RU2780586C2 - Methods and corresponding compositions for manufacture of food products and feeds - Google Patents

Abstract

FIELD: entomology.

SUBSTANCE: present invention relates to a method for improvement of a nutrient profile of an insect by delivery of an effective number of bacteria producing methionine to the insect, wherein the insect is at the stage of larva development.

EFFECT: obtaining compositions for feeding insects.

6 cl, 9 ex, 5 dwg, 12 tbl

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority over U.S. Provisional Application No. 62/450038, filed January 24, 2017, and U.S. Provisional Application No. 62/584011, filed November 9, 2017, the contents of which are incorporated herein by reference in their entirety. volume.

BACKGROUND OF THE INVENTION

Arthropods such as crickets, cicadas, grasshoppers, ants, insect larvae, caterpillars and scorpions have many traditional and potential new uses in the manufacture of food and feed, for humans and animals, respectively. The use of insects as food and feed is becoming a particularly pressing issue due to the increasing cost of animal protein, the insecurity of food and feed, environmental pressures, population growth, and the increasing need for affordable and sustainable sources of nutrients for humans and animals (for example, farm animals). In order to grow beneficial arthropods for use in the food and feed industry, there is a need in the art for methods to promote the growth and fitness of such arthropods.

Brief description of the invention

Disclosed herein are compositions and methods for modulating insect fitness for food and feed production. The composition contains an agent that alters the level, activity, or metabolism of one or more microorganisms living in the host, the change resulting in modulation of the host's fitness.

In one aspect, provided herein is a method for improving the nutritional profile of an insect, the method comprising delivering an effective amount of methionine-producing bacteria to the insect.

In some embodiments, the insect is a cricket, grasshopper, or locust.

In some embodiments, the insect may be at a developmental stage corresponding to an embryo, larva, pupa, or adult.

In some embodiments, delivery may include delivery of the composition to at least one habitat in which the insect grows, lives, breeds, or feeds.

In some embodiments, the methionine-producing bacteria can be delivered in an insect-edible composition for ingestion by insects.

In some embodiments, the methionine-producing bacteria may be formulated with an agriculturally acceptable carrier in the form of a liquid, solid, aerosol, paste, gel, or gaseous composition.

In some embodiments, the carrier may be a seed coater.

In a second aspect, this document provides a modified insect containing exogenous methionine-producing bacteria living in the body of the insect.

In some embodiments of the second aspect, the insect is at a developmental stage corresponding to an embryo, larva, pupa, or adult.

In some embodiments of the second aspect, the methionine-producing bacteria alter the microbiota in the gut and/or hemocoel of the insect.

In yet another aspect, the composition comprises an agent that alters the level, activity, or metabolism of one or more micro-organisms living in the body of an insect host, wherein the change leads to an increase in the fitness of the insect host.

In some embodiments, the implementation of any of the above compositions, one or more microorganisms can be a bacterium or fungus that lives in the host body. In some embodiments, the host bacterium is at least one selected from the group consisting of Candidatus spp, Buchenera spp, Blattabacterium spp, Baumania spp, Wigglesworthia spp, Wolbachia spp, Rickettsia spp, Orientia spp, Sodalis spp, Burkholderia spp, Cupriavidus spp, Frankia spp, Snirhizobium spp, Streptococcus spp, Wolinella spp, Xylella spp, Erwinia spp, Agrobacterium spp, Bacillus spp, Paenibacillus spp, Streptomyces spp, Micrococcus spp, Corynebacterium sppyan, Acetobacter sppyan, Salmonella spp, Rhodococcus spp, Pseudomonas spp, Lactobacillus spp, Enterococcus spp, Alcaligenes spp, Klebsiella spp, Paenibacillus spp, Arthrobacter spp, Corynebacterium spp, Brevibacterium spp, Thermus spp, Pseudomonas spp, Clostridium spp and Escherichia spp. In some embodiments, the host fungus is at least one selected from the group consisting of Candida, Metschnikowia, Debaromyces, Starmerella, Pichia, Cryptococcus, Pseudozyma, Symbiotaphrina bucneri, Symbiotaphrina kochii, Scheffersomyces shehatae, Scheffersomyces stipites , Cryptococcus, Trichosporon, Amylostereum areolatum, Epichloe spp, Pichia pinus, Hansenula capsulate, Daldinia decipien, Ceratocytis spp, Ophiostoma spp and Attamyces bromatificus. In some cases, the bacterium is a naturally occurring bacterium that is capable of producing nutrients (eg, amino acids, eg, methionine).

In any of the above compositions, an agent, which hereinafter may also be referred to as a modulating agent, can alter the growth, division, viability, metabolism, and/or lifespan of a microorganism inhabiting the host organism. In any of the above embodiments, the modulating agent may reduce the viability of one or more microorganisms living in the host. In some embodiments, the implementation of the modulating agent increases the growth or viability of one or more microorganisms living in the host.

In any of the above embodiments, the modulator is a phage, a polypeptide, a small molecule, an antibiotic, a bacterium, or any combination thereof.

In some embodiments, the phage binds to a cell surface protein in a bacterium that lives in the host. In some embodiments, the implementation of the phage is virulent against a bacterium that lives in the host. In some embodiments, the phage is at least one selected from the group consisting of Myoviridae, Siphoviridae, Podoviridae, Lipothrixviridae, Rudiviridae, Ampullaviridae, Bicaudaviridae, Clavaviridae, Corticoviridae, Cystoviridae, Fuselloviridae, Gluboloviridae, Guttaviridae, Inoviridae, Leviviridae, Microviridae , Plasmaviridae and Tectiviridae.

In some embodiments, the polypeptide is at least one of a bacteriocin, an R-type bacteriocin, a C-rich nodule peptide, an antimicrobial peptide, a lysine, or a bacteriocyte regulatory peptide.

In some embodiments, the small molecule is a metabolite.

In some embodiments, the antibiotic is a broad spectrum antibiotic.

In some embodiments, the modulating agent is a naturally occurring bacterium. In some embodiments, the bacterium is at least one bacterium selected from the group consisting of Bartonella apis, Parasaccharibacter apium, Frischella perrara, Snodgrassella alvi, Gilliamela apicola, Bifidobacterium spp , and Lactobacillus spp. In some embodiments, the bacterium is at least one selected from the group consisting of Candidatus spp, Buchenera spp, Blattabacterium spp, Baumania spp, Wigglesworthia spp, Wolbachia spp, Rickettsia spp, Orientia spp, Sodalis spp, Burkholderia spp, Cupriavidus spp, Frankia spp, Snirhizobium spp, Streptococcus spp, Wolinella spp, Xylella spp, Erwinia spp, Agrobacterium spp, Bacillus spp, Paenibacillus spp, Streptomyces spp, Micrococcus spp, Corynebacterium spp, Acetobacter spp, Cyanobacteria spp, Rpphodocuscus, Pseudomonas spp, Lactobacillus spp, Enterococcus spp, Alcaligenes spp, Klebsiella spp, Paenibacillus spp, Arthrobacter spp, Corynebacterium spp, Brevibacterium spp, Thermus spp, Pseudomonas spp, Clostridium spp and Escherichia spp.

In any of the above compositions, host fitness can be measured by host survival, reproduction or metabolism. In some embodiments, the modulating agent modulates host fitness by reducing the host's susceptibility to pesticides (eg, the susceptibility to the pesticide shown in Table 12). In some embodiments, pesticide susceptibility is bactericide or fungicide susceptibility. In some embodiments, pesticide susceptibility is insecticide susceptibility.

In any of the above compositions, the composition may contain a variety of different modulators. In some embodiments, the composition contains a modulating agent and a pesticidal agent (eg, the pesticide listed in Table 12). In some embodiments, the pesticidal agent is a bactericidal or fungicidal agent. In some embodiments, the pesticidal agent is an insecticidal agent.

In any of the above compositions, the modulating agent may be associated with the second moiety. In some embodiments, the second fragment may be a modulator.

In any of the above compositions, the modulator may be associated with a targeting domain. In some embodiments, the targeting domain targets the modulator to a target site in the host. In some embodiments, the targeting domain targets the modulator to one or more microorganisms residing in the host.

In any of the above compositions, the modulating agent may contain an inactivating pre or pro sequence, thereby forming a precursor modulating agent. In some embodiments, the modulator precursor is converted to its active form in the host.

In any of the above compositions, the modulating agent may contain a linker. In some embodiments, the linker is a cleavable linker.

In any of the above compositions, the composition may further comprise a carrier. In some cases, the carrier may be an agriculturally acceptable carrier.

In any of the above compositions, the composition may further comprise a host bait, a sticky agent, or a combination thereof. In some embodiments, the host bait is an edible and/or a chemoattractant.

In any of the above compositions, the composition may be contained in a dosage effective to modulate host fitness.

In any of the above embodiments, the modulating agent in the composition may be effective to increase the production of a nutrient in the host body over a reference level. In some embodiments, the modulating agent is a microorganism that produces a nutrient. In some embodiments, the microorganism is a bacterium. In some embodiments, the nutrient is a vitamin, carbohydrate, amino acid, or polypeptide. In certain embodiments, the amino acid is methionine.

In any of the above compositions, the composition may be formulated for delivery to a microorganism residing in the gut of the host.

In any of the above compositions, the composition may be formulated for delivery to a microorganism residing in the host's bacteriocyte and/or the host's gut. In some embodiments, the composition may be formulated for delivery to a plant. In some embodiments, the composition may be formulated for use in a host feeder.

In any of the above compositions, the composition may be formulated as a liquid, powder, granules or nanoparticles. In some embodiments, the composition is formulated as a composition selected from the group consisting of a liposome, a polymer, a peptide secreting bacterium, and a synthetic nanocapsule. In some embodiments, the synthetic nanocapsule delivers the composition to a target site in the host. In some embodiments, the target site is the gut of the host. In some embodiments, the target site is a bacteriocyte in the host.

In an additional aspect, this document also provides hosts that contain any of the above compositions. In some embodiments, the implementation of the host is an insect. In some embodiments, the insect is an insect belonging to the order Anoplura, Araneae, Blattodea, Coleoptera, Dermaptera, Dictyoptera, Diplura, Diptera, Embioptera, Ephemeroptera, Grylloblatodea, Hemiptera, Homoptera, Hymenoptera, Isoptera, Lepidoptera, Mantodea, Mecoptera, Neuroptera, Odonata, Orthoptera, Phasmida, Plecoptera, Protura, Psocoptera, Siphonaptera, Siphunculata, Thysanura, Strepsiptera, Thysanoptera, Trichoptera or Zoraptera. In certain embodiments, the insect is a cricket. In certain embodiments, the insect is a grasshopper. In some embodiments, the insect is a locust.

In yet another further aspect, this document also provides a system for modulating host fitness, comprising a modulating agent that targets a microorganism required for host fitness, wherein the system is effective to modulate host fitness, and wherein the host is an insect. The modulator may include any of the compositions described herein. In some embodiments, the modulator is formulated as a powder. In some embodiments, the implementation of the modulating agent is in the form of a solvent. In some embodiments, the modulator is formulated as a concentrate. In some embodiments, the implementation of the modulating agent is in the form of a diluent. In some embodiments, the modulator is prepared for delivery by combining any of the previous compositions with a carrier.

In another aspect, this document also provides methods for modulating the fitness of an insect using any of the compositions described herein. In one instance, a method for modulating the fitness of an insect host comprises delivering the composition of any of the preceding paragraphs to the host, wherein the modulating agent targets one or more micro-organisms resident in the host and thereby modulates the fitness of the host. In another case, the method of modulating the diversity of microorganisms in the body of an insect host provides for the delivery of the composition according to any of the previous paragraphs to the host, while the modulating agent purposefully affects one or more microorganisms living in the host body, and thereby modulates the diversity of microorganisms in the host body.

In some embodiments of any of the above methods, the modulating agent may alter the levels of one or more microorganisms living in the host. In some embodiments of any of the above methods, the modulating agent may reduce alter the function of one or more micro-organisms residing in the host. In some embodiments, one or more of the microorganisms may be a bacterium and/or a fungus. In some embodiments, one or more microorganisms are required for host fitness. In some embodiments, one or more microorganisms are required for host survival.

In some embodiments of any of the above methods, the delivery step may include providing the modulator at a dose and for a period of time sufficient to affect one or more microorganisms, thereby modulating microbial diversity in the host. In some embodiments, the delivery step includes topical application of any of the previous compositions to a plant. In some embodiments, the delivery step includes providing a modulating agent via a genetically modified plant. In some embodiments, the delivery step includes providing the modulator to the host as an edible agent. In some embodiments, the delivery step includes obtaining a host carrying a modulator. In some embodiments, the host carrying the modulating agent may transfer the modulating agent to one or more additional hosts.

In some embodiments of any of the above methods, the composition is effective in improving the health and/or survival of the host. In some embodiments, the composition is effective to increase host fitness, increase host longevity, increase effective pollination, increase host product production, increase host breeding, or a combination thereof. In some embodiments, the composition is effective in reducing host susceptibility to a pesticide (eg, the pesticide listed in Table 12). In some embodiments, the pesticidal agent is a neonicotinoid. In some embodiments, the implementation of the composition is effective to increase the resistance of the host to the allelochemical agent produced by the plant. In some embodiments, the allelochemical agent is toxic to the host prior to delivery of the composition. In some embodiments, the allelochemical agent is caffeine, soy cystatin N, monoterpenes, diterpenic acids, or phenolic compounds. In some embodiments, the composition is effective to increase the production of a nutrient in the host body, thereby increasing the nutrient content of the host-derived product. In some embodiments, the nutrient is a vitamin, carbohydrate, amino acid, or polypeptide.

In some embodiments of any of the above methods, at least one portion of the host may be used in the manufacture of a consumable product. In some embodiments of any of the above methods, the host is an insect. In some embodiments, the insect is an insect belonging to the orderAnoplura, Araneae, Blattodea, Coleoptera, Dermaptera, Dictyoptera, Diplura, Diptera, Embioptera, Ephemeroptera, Grylloblatodea, Hemiptera, Homoptera, Hymenoptera, Isoptera, Lepidoptera, Mantodea, Mecoptera, Neuroptera, Odonata, Orthoptera, Phasmida, Plecoptera, Procoptera, Psocoptera, Siphonaptera, Siphunculata, Thysanura, Strepsiptera, Thysanoptera, TrichopteraorZoraptera.In certain embodiments, the insect is a cricket. In certain embodiments, the insect is a grasshopper. In some embodiments, the insect is a locust. In some embodiments, the product includes a human food product. In some embodiments, the product includes a nutritional supplement that complements an animal food or human food product. In some embodiments, the product includes pet food. In some embodiments, the animals are livestock or farm animals.

In another aspect, provided herein is a method for producing a food product for a human or animal that includes (a) providing a plurality (e.g., 2, >2, >5, >10, >100, >1000, >5000, >10000, > 50,000, >100,000) insect hosts, (b) delivering an ingestible composition described herein to a large number of insect hosts in an amount effective to modulate one or more micro-organisms inhabiting the insect host from a plurality of and ( c) processing the plurality (eg, grinding, optionally mixing with a carrier or other food component) into a food product, feed additive or nutritional supplement. In some embodiments, the ingestible composition comprises a microorganism. In some embodiments, the implementation of the microorganism produces a nutrient and the microorganism is effective to increase the production of the nutrient in the host compared to the reference level. In some embodiments, the microorganism is a bacterium. In some embodiments, the nutrient is a vitamin, carbohydrate, amino acid, or polypeptide. In some embodiments, the amino acid is methionine.

In some embodiments of any of the above methods, the delivery step includes delivering any of the previously prepared compositions to a plant. In some embodiments, the plant is a crop. In some embodiments, the crop is an unharvested crop at the time of delivery. In some embodiments, the crop is the harvested crop at the time of delivery. In some embodiments, the crop includes harvested fruits or vegetables. In some embodiments, the composition is delivered in an amount and for a period of time effective to enhance crop growth. In some embodiments, the crop includes corn, soybean, or wheat plants.

In another aspect, this document also provides screening assays to identify a modulator that modulates host fitness. In one instance, a screening assay to identify a modulator that modulates host fitness includes the steps of (a) exposing a microorganism that may inhabit the host to one or more candidate modulators, and (b) identifying a modulator that increases or decreases fitness. owner.

In some embodiments of the screening assay, the modulating agent is a microorganism that lives in the host. In some embodiments, the microorganism is a bacterium. In some embodiments, the implementation of the bacterium, living in the host organism, increases the fitness of the host. In some embodiments, the implementation of the bacterium breaks down the pesticide (for example, the pesticide shown in table 12). In some embodiments, the pesticide is a neonicotinoid. In some embodiments, the bacterium secretes an amino acid. In some embodiments, the amino acid is methionine.

In some embodiments of the screening assay, the modulating agent adversely affects the microorganism that degrades the allelochemical. In some embodiments, the modulating agent is a phage, an antibiotic, or a test compound. In some embodiments, the antibiotic is thimentin or azithromycin.

In some embodiments of the screening assay, the host may be an invertebrate. In some embodiments, the invertebrate is an insect. In some embodiments, the insect is a cricket. In certain embodiments, the insect is a grasshopper. In some embodiments, the insect is a locust.

In any of the above screening assay embodiments, host fitness can be modulated by modulating the host microbiota.

Definitions

Used in this document, the term "bacteriocin" refers to a peptide or polypeptide that has antimicrobial properties. Naturally occurring bacteriocins are produced by certain prokaryotes and act against organisms related to the producer strain, but not against the producer strain itself. The bacteriocins provided herein include, without limitation, naturally occurring bacteriocins, such as bacteriocins produced by bacteria, and derivatives thereof, such as engineered bacteriocins, recombinantly expressed bacteriocins, and chemically synthesized bacteriocins. In some instances, the bacteriocin is a functionally active variant of the bacteriocins described herein. In some cases, the bacteriocin variant is at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83 %, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical , for example, in a certain area or in the entire sequence, the sequence of the bacteriocin described herein, or the naturally occurring bacteriocin.

As used herein, the term "bacteriocyte" refers to a specialized cell found in certain insects that is inhabited by intracellular bacteria with the properties of symbiotic bacteria.

As used herein, the term "effective amount" refers to an amount of a modulating agent (e.g., phage, lysin, bacteriocin, small molecule, or antibiotic) or composition containing said agent, sufficient to achieve the specified result, for example, to increase or promote the fitness of an organism - host (eg, insect); to achieve a target level (eg, a predetermined or threshold level) of the concentration of the modulating agent in the body of the target host; to achieve a target level (eg, a predetermined or threshold level) of the concentration of the modulating agent in the gut of the target host; to achieve a target level (eg, a predetermined or threshold level) of the concentration of the modulating agent in the bacteriocyte of the target host; to modulate the level or activity of one or more microorganisms (eg, endosymbionts) in the target host.

As used herein, the term “fitness” refers to the ability of a host organism to survive and/or produce viable offspring. The fitness of an organism can be measured by one or more parameters, including but not limited to lifespan, nutrient production, reproduction rate, motility, body weight, and metabolic rate. Fitness can be further measured based on activity measurements or product yield.

As used herein, the term "gut" refers to any part of the host's gut, including the foregut, midgut, or hindgut of the host.

As used herein, the term "host" refers to an organism (e.g., an insect) hosting microorganisms (e.g., endogenous microorganisms, endosymbiotic microorganisms (e.g., primary or secondary endosymbionts), commensal organisms, and/or pathogenic microorganisms).

As used herein, the term "enhancing host fitness" or "promoting host fitness" refers to any beneficial change in host physiological processes or any activity carried out by said host due to the administration of a modulating agent, including, without limitation, any one or more of the following desired effects. : (1) an increase in the host population of about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 99%, 100% or more; (2) increasing the reproduction rate of the host (eg, insect) by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 99%, 100%, or more; (3) increasing host (eg, insect) motility by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 99%, 100% or more ; (4) increasing the body weight of the host (eg, insect) by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 99%, 100%, or more; (5) increasing the metabolic rate or activity of the host (eg, insect) by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 99%, 100 % or more; (6) increasing the production of host by-products by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 99%, 100% or more; (7) increasing the content of a host (eg, insect) nutrient (eg, protein, fatty acids, or amino acids) by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90 %, 95%, 99%, 100% or more; or (8) increasing host resistance to the pesticide by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 99%, 100% or more. An increase in host fitness can be measured in comparison to a host that has not been administered a modulating agent.

The term "insect" includes any organism belonging to the phylum Arthropoda and the class Insecta or the class Arachnida at any stage of development, i.e. immature and adult insects.

As used herein, the term "lysin", also known as endolysin, autolysin, murein hydrolase, peptidoglycan hydrolase, or cell wall hydrolase, refers to a hydrolytic enzyme that can lyse a bacterium by cleaving the peptidoglycan in the bacterial cell wall. The lysins provided herein include, without limitation, naturally occurring lysins such as phage-produced lysins, bacterial-produced lysins, and derivatives thereof, such as engineered lysins, recombinantly expressed lysins, and chemically synthesized lysins. A functionally active bacteriocin variant may be at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83 %, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical , for example, in a certain area or in the entire sequence, the sequence of a synthetic, recombinant or naturally occurring bacteriocin, including any described herein.

As used herein, the term "microorganism" refers to bacteria or fungi. Microorganisms can refer to microorganisms living in the host organism (for example, endogenous microorganisms, endosymbiotic microorganisms (for example, primary or secondary endosymbionts)), or microorganisms exogenous to the host, including microorganisms that can act as modulators. As used herein, the term “target microorganism” refers to a microorganism that resides in the host organism and is directly or indirectly exposed to the modulating agent.

As used herein, the term “modulating agent” or “agent” refers to an agent that is capable of altering the levels and/or functioning of microorganisms living in a host organism (eg, an insect) and thereby modulating (eg, increasing) the organism's fitness host (for example, an insect).

As used herein, the term “improving the nutritional profile of an insect” refers to an increase in nutrient production that can increase the protein content, body weight, and/or overall nutritional value of an insect.

As used herein, the term "pesticide" or "pesticidal agent" refers to a substance that can be used to control pests of agricultural, environmental and household/domestic significance such as insects, fungi, bacteria or viruses. The term “pesticide” is understood to encompass naturally occurring or synthetic insecticides (larvicides or adulticides), insect growth regulators, acaricides (miticides), nematicides, ectoparasiticides, bactericides, fungicides or herbicides (substances that can be used in agriculture to control or modify plant growth). Additional examples of pesticides or pesticides are shown in Table 12. In some cases, the pesticide is an allelochemical. As used herein, the term “allelochemical” or “allelochemical agent” means a substance produced by an organism that can affect the physiological function (eg, inception, growth, survival, or reproduction) of another organism (eg, insect host).

As used herein, the term "peptide", "protein", or "polypeptide" encompasses any chain of naturally occurring or non-naturally occurring amino acids (both D- and L-amino acids), regardless of length (e.g., at least 2, 3, 4, 5, 6, 7, 10, 12, 14, 16, 18, 20, 25, 30, 40, 50, 100 or more amino acids), the presence or absence of post-translational modifications (eg, glycosylation or phosphorylation) or having, for example, one or more non-aminoacyl groups (e.g., carbohydrate, lipid, etc.) covalently linked to the peptide, and includes, for example, naturally occurring proteins, synthetic or recombinant polypeptides and peptides, hybrid molecules, peptoids or peptidomimetics.

As used herein, "percent identity" between two sequences is determined using the BLAST 2.0 algorithm as described in Altschul et al., (1990) J. Mol. Biol. 215:403-410. BLAST analysis software is publicly available through the National Center for Biotechnology Information.

As used herein, the term "bacteriophage" or "phage" refers to a virus that infects and replicates in bacteria. Bacteriophages replicate in bacteria after the introduction of their genome into the cytoplasm and do so through a lytic cycle, which results in the lysis of the bacterial cell, or a lysogenic (non-lytic) cycle, in which the bacterial cell remains intact. The phage may be a naturally occurring phage isolate or engineered phage, including vectors or nucleic acids that encode a partial genome of the phage (e.g., including at least all of the essential genes required for the life cycle of the phage in a host bacterium) or the complete phage genome.

As used herein, the term "plant" refers to whole plants, plant organs, plant tissues, plant cells, seeds, or progeny thereof. Plant cells include, without limitation, cells from seeds, suspension cultures, embryos, meristem patches, callus tissue, leaves, roots, shoots, gametophytes, sporophytes, pollen, or microspores. Plant parts include differentiated or undifferentiated tissues, including, but not limited to, roots, stems, shoots, leaves, pollen, seeds, tumor tissue, and various cell and culture forms (eg, single cells, protoplasts, embryos, or callus tissue). The plant tissue may be in a plant or in an organ, tissue or cell culture of a plant. In addition, the plant may be genetically modified such that it produces a heterologous protein or RNA, for example, any of the modulators in the methods and compositions described herein.

The terms “obtainable”, “producible”, or the like are used to indicate that a claim or embodiment refers to a compound, composition, product, etc. per se, i.e. that the compound, composition, product, etc. can be obtained or produced using the method that is described for the manufacture of a compound, composition, product, etc., except that the compound, composition, product, etc. can also be obtained or produced using methods other than those described. The terms "obtained", "produced" or the like indicate that the connection, composition, product receive or produce using a particular specified method. It should be understood that the terms “obtained”, “produced”, etc. also disclose the terms “obtained”, “produced”, etc. as a preferred embodiment, “obtained”, “produced”, etc.

Other characteristics and advantages of the present invention will be apparent from the following detailed description and claims.

BRIEF DESCRIPTION OF GRAPHICS

The figures are provided to illustrate one or more characteristics, aspects or embodiments of the present invention and are not intended to be limiting.

In FIG. 1 is a graph showing the time to mature from an embryo in Drosophila melanogaster . Drosophila melanogaster embryos were grown either with a diet inoculated with Corynebacterium glutamicum (a strain that produces glutamate - C. glutamicum Glu) or with an axenic diet without any bacterium. The percentage of adults emerging from their pupa was measured every 12 hours from the time of first adult appearance. Organisms grown on a bacteria-supplemented diet reach adulthood faster than their bacteria-free counterparts.

In FIG. 2A is a graph showing the effects of males on developmental rate differences in Drosophila melanogaster . The sex of adults was determined, the appearance of which is shown in FIG. 1, and the rate of their appearance was displayed on the graph.

In FIG. 2B is a graph showing the effects of females on developmental rate differences in Drosophila melanogaster . The sex of adults was determined, the appearance of which is shown in FIG. 1, and the rate of their appearance was displayed on the graph. The increase in the rate of development in females due to the presence of bacteria in the diet is significantly greater than that of their male counterparts. The beneficial effects of the presence of bacteria in the diet of flies are more pronounced in females than in males.

In FIG. 3 is a graph showing the strain of C. glutamicum that contributes to the increase in larval biomass. Larvae reared on a diet supplemented with strains of C. glutamicum that produce either glutamate or methionine are larger than larvae reared on a sterile diet or a diet supplemented with Escherichia coli . Larval areas are measured as the number of pixels in the larval images. Medians and 95% confidence intervals are shown as lines on the graph.

In FIG. 4 is a panel of graphs showing the results of Seahorse extracellular flux analysis for bacterial respiration. Bacteria were grown to logarithmic phase and loaded onto Seahorse XFe96 plates for temporal measurements of oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) as described in the methods. The treatments were injected into the wells after about 20 minutes and the bacteria were monitored for changes in growth. Rifampicin=100 µg/ml; chloramphenicol=25 µg/ml; phages (T7 in the case of E. coli and ΦSmVL-C1 in the case of Serratia marcescens ) were lysates diluted 1:2 or 1:100 in SM buffer. The marks on each line are provided solely as indicators of the status to which each line corresponds and do not indicate data points.

In FIG. 5 is a graph showing that the phage against S. marcescens led to a reduction in fly mortality. All flies injected with S. marcescens by puncture died within a day, while a significant proportion of flies injected with both S. marcescens and phage by puncture survived for five days after treatment. Almost all of the control flies, which were not treated in any way, survived until the end of the experiment. A log-rank test was used to compare curves to determine statistical significance, an asterisk indicates p<0.0001.

DETAILED DESCRIPTION OF THE INVENTION

Provided herein are methods and compositions for food and feed applications, such as altering the level, activity, or metabolism of one or more micro-organisms residing in an insect host, the change resulting in increased host fitness. The present invention describes a composition that contains a modulating agent (eg, phage, peptide, small molecule, antibiotic, or combinations thereof) that can alter the host's microbiota in a manner that is beneficial to the host. The modulator described herein can be used to improve fitness of a range of insects used in the human and animal food industry by promoting favorable microbial levels, microbial activity, microbial metabolism, and/or microbial diversity.

The methods and compositions described herein are based, in part, on examples that illustrate how various agents, such as methionine-producing microorganisms, can be used on insect hosts such as cricket, fly, grasshopper, or locust to indirectly improve health (eg, increase in methionine content, body weight, growth rate and/or survival) of these hosts by changing the level, activity or metabolism of microorganisms in these hosts. Methionine-producing microorganisms are an illustrative example of amino acid-producing microorganisms, and are generally illustrative nutrient-producing microorganisms, and other microorganisms of this type can be used in the present invention. Based on this, the present disclosure describes a number of different approaches for the use of agents that alter the level, activity or metabolism of one or more microorganisms living in the host, the change resulting in modulation of the fitness of the host.

hosts

Insects

The host in any of the compositions or methods described herein can be any organism belonging to the phylum Arthropoda (eg, insects), including any of the arthropods described herein. In some cases, the host may be an insect or arachnid that may be reared for the consumable product (eg food or feed). For example, the host may be a moth, butterfly, fly, cricket, grasshopper, locust, spider, or beetle. In some cases, the host belongs to the order Anoplura, Araneae, Blattodea, Coleoptera, Dermaptera, Dictyoptera, Diplura, Diptera, Embioptera, Ephemeroptera, Grylloblatodea, Hemiptera, Homoptera, Hymenoptera, Isoptera, Lepidoptera, Mantodea, Mecoptera, Neuroptera, Odonata, Orthoptera, Phasmida , Plecoptera, Protura, Psocoptera, Siphonaptera, Siphunculata, Thysanura, Strepsiptera, Thysanoptera, Trichoptera or Zoraptera .

In some examples, the host is the black soldier fly ( Hermetia illucens ), housefly, pitch-brown beetle, tailor ant, silkworm ( Bombyx mori ), grasshopper, Chinese grasshopper ( Acrida cinerea ), flour beetle ( Clarias gariepinns ), moth ( Anaphe infracta or Bombyx mori ), Spodoptera littoralis , house cricket, termite, palm weevil ( Rhynchophorus ferruginens ), belostomatid ( Lethocerus indicus ), water beetle, termite ( Macrotermes subhyalinus ), bread grinder ( Stegobium paniceum ), Imbrasia belina, Rhynchophorus phoenicis , Oryctes rhinoceros, Macrotermes bellicosus, Ruspolia differens , Oryctes Monoceros or Oecophylla smaragdina.

In specific cases, the modulators disclosed herein can be used to improve the fitness of crickets, grasshoppers or locusts.

The host can be at any stage of development. For example, the host may be an embryo, larva, pupa, or adult.

Host Fitness

The methods and compositions provided herein can be used to improve fitness in any of the hosts described herein. An increase in fitness may result from any change in the microorganisms living in the host organism, the change being a consequence of the administration of the modulating agent and conferring beneficial or beneficial effects on the host.

In some cases, an increase in host fitness may be manifested as an improvement in the physiological processes of the host (eg, improved health or survival) due to the administration of a modulating agent. In some cases, the fitness of an organism may be measured by one or more parameters, including but not limited to reproduction rate, lifespan, motility, fecundity, body weight, metabolic rate, or activity or survival compared to a host organism in which the modulating agent is not was entered. For example, the methods or compositions provided herein may be effective in improving the general health of the host or in improving the overall survival of the host compared to a host to which a modulating agent has not been administered. In some cases, the improvement in host survival is approximately 2%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100% or more than 100% compared with a reference level (eg, the level found in a host that does not receive modulator). In some cases, the methods and compositions are effective in increasing the intensity of reproduction of the host (eg, the rate of reproduction) compared to the host organism, which has not been administered modulating agent. In some cases, the methods and compositions are effective in increasing other physiological parameters, such as mobility, body weight, lifespan, fertility, or metabolic rate, by about 2%, 5%, 10%, 20%, 30%, 40%, 50 %, 60%, 70%, 80%, 90%, 100%, or more than 100% compared to the reference level (eg, the level found in a host that is not receiving a modulating agent).

In some cases, increased host fitness may manifest as increased production of one or more nutrients in the host (eg, vitamins, carbohydrates, amino acids, or polypeptides) compared to a host that has not been administered a modulating agent. In some cases, the methods or compositions provided herein may be effective to increase the production of nutrients in the host body (eg, vitamins, carbohydrates, amino acids or polypeptides) by about 2%, 5%, 10%, 20%, 30% , 40%, 50%, 60%, 70%, 80%, 90%, 100%, or more than 100% compared to the reference level (eg, the level found in a host that does not receive a modulating agent). In some instances, the methods or compositions provided herein can increase the nutrient content of a host by increasing the nutrient production of one or more microorganisms (e.g., endosymbionts) in the host compared to a host that has not been administered a modulating agent. .

In some cases, an increase in host fitness may manifest itself as a decrease in the susceptibility of the host to a pesticide (for example, the pesticide listed in Table 12) and / or an increase in host resistance to the pesticide (for example, the pesticide listed in Table 12) compared to the organism - a host to which the modulating agent has not been administered. In some instances, the methods and compositions provided herein may be effective in reducing a host's sensitivity to a pesticide by about 2%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70% , 80%, 90%, 100%, or more than 100% compared to the reference level (eg, the level found in a host that is not receiving a modulating agent). The pesticidal agent may be any pesticidal agent known in the art, including insecticidal agents. In some cases, the pesticidal agent is a neonicotinoid. In some instances, the methods or compositions provided herein may desensitize a host to a pesticide (e.g., the pesticide listed in Table 12) by increasing the host's ability to metabolize or break down the pesticide into usable substrates relative to the host organism. to which the modulating agent has not been administered.

In some cases, an increase in host fitness may manifest itself as a decrease in the sensitivity of the host to the allelochemical agent and/or an increase in the resistance of the host to the allelochemical agent compared to a host organism to which the modulating agent has not been introduced. In some instances, the methods or compositions provided herein may be effective in increasing host resistance to an allelochemical agent by about 2%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70% , 80%, 90%, 100%, or more than 100% compared to the reference level (eg, the level found in a host that does not receive a modulator). In some cases, the allelochemical agent is caffeine, soy cystatin N, monoterpenes, diterpenic acids, or phenolic compounds compared to a host organism to which the modulating agent has not been administered. In some instances, the methods or compositions provided herein can desensitize a host to an allelochemical agent by increasing the host's ability to metabolize or break down the allelochemical agent into usable substrates compared to a host that has not received the modulating agent.

In some instances, the methods or compositions provided herein may be effective in increasing host resistance to parasites or pathogens (e.g., fungal, bacterial, or viral pathogens; or parasitic microscopic mites) compared to a host that has not had a modulating agent. entered. In some instances, the methods or compositions provided herein may be effective in increasing host resistance to a pathogen or parasite (e.g., fungal, bacterial, or viral pathogens; or parasitic microscopic mites) by about 2%, 5%, 10%, 20 %, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, or greater than 100% compared to the reference level (e.g., the level found in a host that is not receiving modulator ).

In some cases, increased host fitness may manifest itself as other fitness benefits, such as improved tolerance for certain environmental factors (for example, tolerance for high or low temperatures), improved ability to survive in certain environments, or improved ability to maintain a certain diet (for example, improved ability to metabolize soy compared to corn) compared to a host that has not received the modulating agent. In some instances, the methods or compositions provided herein may be effective in improving host fitness in a variety of ways described herein. In addition, the modulating agent can increase host fitness in any number of host classes, orders, families, genera, or species (e.g., 1 host species, 2, 3, 4, 5, 6, 7, 8, 9,10, 15, 20, 30, 40, 50, 60, 70, 80, 90, 100, 150, 200, 200, 250, 500 or more host species). In some cases, the modulating agent acts on one host class, order, family, genus, or species.

Host fitness can be assessed using any of the standard methods in the art. In some cases, host fitness can be assessed using an individual host score. Alternatively, host fitness can be assessed by assessing the host population. For example, an increase in host fitness may manifest itself as an increase in successful competition relative to other insects, thereby leading to an increase in the size of the host population.

Insect Hosts in Feed/Food Manufacturing

Once the desired stage of development has been reached, the host can be harvested and, if necessary, processed for use in the manufacture of a consumable product. In some cases, the harvested insect may be distributed in whole form (eg, whole, unprocessed insect) as a consumable product. In some cases, the whole harvested insect is processed (eg ground) and distributed as a consumable product. Alternatively, one or more parts of the insect (eg, one or more body parts or one or more substances) can be removed from the insect for use in the manufacture of a consumable product.

The consumable product can be any product that is safe for human or animal consumption (eg, ingestion). In some cases, the host may be used in the manufacture of animal feed. In some instances, the animal is a livestock or farm animal (eg, chicken, cow, horse, or pig). In some cases, the animal is a bird, reptile, amphibian, mammal, or fish. In some cases, the host may be used in the manufacture of a product that replaces the conventional pet food. Alternatively, the host may be used in the manufacture of a product that complements a conventional pet food. The host may also be used in the manufacture of a human food, feed additive or food ingredient. In some cases, the host is used in the manufacture of a nutritional supplement (eg, protein supplement) for humans.

The host may be a wild-type or a domesticated insect. Additionally, the host may be at any stage of development at the time of delivery or use of the compositions described herein. Additionally, the host may be at any stage of development at the time of collection of the host for use in the manufacture of a consumable product. In some cases, the host is a larva, pupa, or adult at the time of collection, use, processing, or manufacture. Delivery of the modulator and the collection steps may occur at the same time or at different times.

In some cases, insect species are selected based on their natural nutritional profile. In some cases, the modulating agent is used to improve the nutritional profile of an insect, where the modulating agent results in increased nutrient production compared to a host that has not received the modulating agent. Examples of nutrients include vitamins, carbohydrates, amino acids, polypeptides or fatty acids. In some cases, the increase in production may result from an increase in the production of the nutrient by the host microorganism. Alternatively, the increase in production may result from an increase in the production of the nutrient by the insect host itself, where the host has increased fitness following delivery or administration of the modulating agent.

In some cases, the final treatment combines a first insect species with a second insect species whose nutritional profile provides a complementary beneficial effect on the overall nutritional value of the food or feed. For example, a species having a high protein profile can be combined with a species having a high omega-3/6 fatty acid profile. Thus, the insect protein feed mixture can be custom-blended to meet the needs of humans or different animal species.

Target microorganisms

Microorganisms targeted by the modulator described herein may include any microorganism residing in or on the host body, including, without limitation, any bacteria and/or fungi described herein. Microorganisms inhabiting the host may include, for example, symbiotic microorganisms (eg, endosymbiotic microorganisms that provide beneficial nutrients or enzymes to the host), commensal microorganisms, pathogenic or parasitic microorganisms. The symbiotic microorganism (eg, bacterium or fungus) may be an obligate host symbiont or a facultative host symbiont. Microorganisms inhabiting the host organism can be acquired by any transfer route, including vertical, horizontal or multiple origin transfer.

bacteria

Exemplary bacteria that can be targeted according to the methods and compositions provided herein include, without limitation , Xenorhabdus spp, Photorhabdus spp, Candidatus spp, Buchnera spp, Blattabacterium spp, Baumania spp, Wigglesworthia spp, Wolbachia spp, Rickettsia spp, Orientia spp, Sodalis spp, Burkholderia spp, Cupriavidus spp, Frankia spp, Snirhizobium spp, Streptococcus spp, Wolinella spp, Xylella spp, Erwinia spp, Agrobacterium spp, Bacillus spp, Paenibacillus spp, Streptomyces spp, Micrococcus spp, Corynebacterium spp, Acetobacter spp, Cyanobacteria spp, Salmonella spp, Rhodococcus spp, Pseudomonas spp, Lactobacillus spp, Enterococcus spp, Alcaligenes spp, Klebsiella spp, Paenibacillus spp, Arthrobacter spp, Corynebacterium spp, Brevibacterium spp, Thermus spp, Pseudomonas spp, Clostridium spp и Escherichia spp . Non-limiting examples of bacteria that can be targeted using the methods and compositions provided herein are shown in Table 1.

Table 1. Examples of target bacteria and insect hosts

Primary endosymbiont Master Location 16S rRNA Gamma proteobacteria Carsonella ruddii Psylloidea (Psylloidea) Bacteriocytes TATCCAGCCACAGGTTCCCCTACAGCTACCTTGTTACGACTTCACCCCAGTTACAAATCATACCGTTGTAATAGTAAAATTACTTATGATACAATTTACTTCCATGGTGTGACGGGCGGTGTGTACAAGGCTCGAGAACGTATTCACCGTAACATTCTGATTTACGATTACTAGCGATTCCAACTTCATGAAATCGAGTTACAGATTTCAATCCGAACTAAGAATATTTTTTAAGATTAGCATTATGTTGCCATATAGCATATAACTTTTTGTAATACTCATTGTAGCACGTGTGTAGCCCTACTTATAAGGGCCATGATGACTTGACGTCGTCCTCACCTTCCTCCAATTTATCATTGGCAGTTTCTTATTAGTTCTAATATATTTTTAGTAAAATAAGATAAGGGTTGCGCTCGTTATAGGACTTAACCCAACATTTCACAACACGAGCTGACGACAGCCATGCAGCACCTGTCTCAAAGCTAAAAAAGCTTTATTATTTCTAATAAATTCTTTGGATGTCAAAAGTAGGTAAGATTTTTCGTGTTGTATCGAATTAAACCACATGCTCCACCGCTTGTGCGAGCCCCCGTCAATTCATTTGAGTTTTAACCTTGCGGTCGTAATCCCCAGGCGGTCAACTTAACGCGTTAGCTTTTTCACTAAAAATATATAACTTTTTTTCATAAAACAAAATTACAATTATAATATTTAATAAATAGTTGACATCGTTTACTGCATGGACTACCAGGGTATCTAATCCTGTTTGCTCCCCATGCTTTCGTGTATTAGTGTCAGTATTAAAATAGAAATACGCCTTCGCCACTAGTATTCTTTCAGATATCTAAGCATTTCACTGCTACTCCTGAAATTCTAATTTCTTCTTTTATACTCAAGTTTATAAGTATTAATTTCAATATTAAATTACTTTAATAAATTTAAAAATTAATTTTTAAAAACAACCTGCACACCCTTTACGCCCAATAATTCCGATTAACGC TTGCACCCCTCGTATTACCGCGGCTGCTGGCACGAAGTTAGCCGGTGCTTCTTTTACAAATAACGTCAAAGATAATATTTTTTTATTATAAAATCTCTTCTTACTTTGTTGAAAGTGTTTTACAACCCTAAGGCCTTCTTCACACACGCGATATAGCTGGATCAAGCTTTCGCTCATTGTCCAATATCCCCCACTGCTGCCTTCCGTAAAAGTTTGGGCCGTGTCTCAGTCCCAATGTGGTTGTTCATCCTCTAAGATCAACTACGAATCATAGTCTTGTTAAGCTTTTACTTTAACAACTAACTAATTCGATATAAGCTCTTCTATTAGCGAACGACATTCTCGTTCTTTATCCATTAGGATACATATTGAATTACTATACATTTCTATATACTTTTCTAATACTAATAGGTAGATTCTTATATATTACTCACCCGTTCGCTGCTAATTATTTTTTTAATAATTCGCACAACTTGCATGTGTTAAGCTTATCGCTAGCGTTCAATCTGAGCTATGATCAAACTCA
(SEQ ID NO: 1) Portiera aleyrodidarum BT-B Whiteflies (Aleyrodoidea) Bacteriocytes
(SEQ ID NO: 2) Buchnera aphidicola, APS strain (Acyrthosiphon pisum) Aphids (Aphidoidea) Bacteriocytes
(SEQ ID NO: 3) Buchnera aphidicola, strain Sg (Schizaphis graminum) Aphids (Aphidoidea) Bacteriocytes
(SEQ ID NO: 4) Buchnera aphidicola strain Bp (Baizongia pistaciae) Aphids (Aphidoidea) Bacteriocytes
(SEQ ID NO: 5) Buchnera aphidicola BCc Aphids (Aphidoidea) Bacteriocytes
(SEQ ID NO: 6) Buchnera aphidicola (Cinara tujafilina) Aphids (Aphidoidea) Bacteriocytes
(SEQ ID NO: 7) Buchnera aphidicola strain G002 (Myzus persicae) Aphids (Aphidoidea) Bacteriocytes ATGAAAAGTATAAAAACTTTTAAAAAACACTTTCCTGTGAAAAAATATGGACAAAATTTTCTTATTAATAAAGAGATCATAAAAAATATTGTTAAAAAAATTAATCCAAATATAGAACAAACATTAGTAGAAATCGGACCAGGATTAGCTGCATTAACTGAGCCCATATCTCAGTTATTAAAAGAGTTAATAGTTATTGAAATAGACTGTAATCTATTATATTTTTTAAAAAAACAACCATTTTATTCAAAATTAATAGTTTTTTGTCAAGATGCTTTAAACTTTAATTATACAAATTTATTTTATAAAAAAAATAAATTAATTCGTATTTTTGGTAATTTACCATATAATATCTCTACATCTTTAATTATTTTTTTATTTCAACACATTAGAGTAATTCAAGATATGAATTTTATGCTTCAAAAAGAAGTTGCTGCAAGATTAATTGCATTACCTGGAAATAAATATTACGGTCGTTTGAGCATTATATCTCAATATTATTGTGATATCAAAATTTTATTAAATGTTGCTCCTGAAGATTTTTGGCCTATTCCGAGAGTTCATTCTATATTTGTAAATTTAACACCTCATCATAATTCTCCTTATTTTGTTTATGATATTAATATTTTAAGCCTTATTACAAATAAGGCTTTCCAAAATAGAAGAAAAATATTACGTCATAGTTTAAAAAATTTATTTTCTGAAACAACTTTATTAAATTTAGATATTAATCCCAGATTAAGAGCTGAAAATATTTCTGTTTTTCAGTATTGTCAATTAGCTAATTATTTGTATAAAAAAAATTATACTAAAAAAAATTAA
(SEQ ID NO: 8) Buchnera aphidicola, strain Ak (Acyrthosiphon kondoi) Aphids (Aphidoidea) Bacteriocytes
(SEQ ID NO: 9) Buchnera aphidicola strain Ua (Uroleucon ambrosiae) Aphids (Aphidoidea) Bacteriocytes
(SEQ ID NO: 10) Buchnera aphidicola (Aphis glycines) Aphids (Aphidoidea) Bacteriocytes
(SEQ ID NO: 11) Annandia Pinicola (Phylloxeroidea) Bacteriocytes
(SEQ ID NO: 12) Moranella endobia (Coccoidea) Bacteriocytes
(SEQ ID NO: 13) Ishikawaella capsulata Mpkobe (Heteroptera) Bacteriocytes
(SEQ ID NO: 14) Baumannia cicadellinicola Leafhoppers (Cicadellinae) Bacteriocytes
(SEQ ID NO: 15) Sodalis-like symbionts Rhopalus sapporensis Broader tissue tropism
(SEQ ID NO: 16) Hartigia pinicola Hermes pine wood Bacteriocytes
(SEQ ID NO: 17) Wigglesworthia glossinidia Tsetse fly (Diptera: Glossinidae) Bacteriocytes Beta Proteobacteria Tremblaya phenacola Phenacoccus avenae (TPPAVE). Bacteriomes
(SEQ ID NO: 18) Tremblaya princeps Grape mealybug Planococcus citri Bacteriomes
(SEQ ID NO: 19) Vidania Bacteriomes Nasuia deltocephalinicola Harmful host insect Macrosteles quadripunctulatus (Hemiptera: Cicadellidae) Bacteriomes
(SEQ ID NO: 20) Zinderia insecticola CARI Pennitsa Clastoptera arizonana Bacteriocytes
(SEQ ID NO: 21) Profftella armatura Diaphorina citri , Asian citrus psyllid Bacteriomes Alpha Proteobacteria Hodgkinia Cicada
Diceroprocta semicincta Bacteriome
(SEQ ID NO: 22) Wolbachia sp. wPip Mosquito
Culex quinquefasciatus Bacteriome
(SEQ ID NO: 23) Bacteroidetes Uzinura diaspidicola Shchitovki Bacteriocytes
(SEQ ID NO: 24) Sulcia muelleri Blue-green leafhoppers and several other leafhopper species Bacteriocytes
(SEQ ID NO: 25) Yeast-like symbionts Symbiotaphrina buchneri , control strain JCM9740 Grinder beetles Stegobium paniceum Mycetoma between the anterior and midgut AGATTAAGCCATGCAAGTCTAAGTATAAGNAATCTATACNGTGAAACTGCGAATGGCTCATTAAATCAGTTATCGTTTATTTGATAGTACCTTACTACATGGATAACCGTGGTAATTCTAGAGCTAATACATGCTAAAAACCCCGACTTCGGAAGGGGTGTATTTATTAGATAAAAAACCAATGCCCTTCGGGGCTCCTTGGTGATTCATGATAACTTAACGAATCGCATGGCCTTGCGCCGGCGATGGTTCATTCAAATTTCTGCCCTATCAACTTTCGATGGTAGGATAGTGGCCTACCATGGTTTTAACGGGTAACGGGGAATTAGGGTTCGATTCCGGAGAGGGAGCCTGAGAAACGGCTACCACATCCAAGGAAGGCAGCAGGCGCGCAAATTACCCAATCCCGACACGGGGAGGTAGTGACAATAAATACTGATACAGGGCTCTTTTGGGTCTTGTAATTGGAATGAGTACAATTTAAATCCCTTAACGAGGAACAATTGGAGGGCAAGTCTGGTGCCAGCAGCCGCGGTAATTCCAGCTCCAATAGCGTATATTAAAGTTGTTGCAGTTAAAAAGCTCGTAGTTGAACCTTGGGCCTGGCTGGCCGGTCCGCCTAACCGCGTGTACTGGTCCGGCCGGGCCTTTCCTTCTGGGGAGCCGCATGCCCTTCACTGGGTGTGTCGGGGAACCAGGACTTTTACTTTGAAAAAATTAGAGTGTTCAAAGCAGGCCTATGCTCGAATACATTAGCATGGAATAATAGAATAGGACGTGCGGTTCTATTTTGTTGGTTTCTAGGACCGCCGTAATGATTAATAGGGATAGTCGGGGGCATCAGTATTCAATTGTCAGAGGTGAAATTCTTGGATTTATTGAAGACTAACTACTGCGAAAGCATTTGCCAAGGATGTTTTCATTAATCAGTGAACGAAAGTTAGGGGATCGAAGACGATCAGATACCGTCGTAGTCTTAACCATAAACTATGCCGACTAG GGATCGGGCGATGTTATTATTTTGACTCGCTCGGCACCTTACGAGAAATCAAAGTCTTTGGGTTCTGGGGGGAGTATGGTCGCAAGGCTGAAACTTAAAGAAATTGACGGAAGGGCACCACCAGGAGTGGAGCCTGCGGCTTAATTTGACTCAACACGGGGAAACTCACCAGGTCCAGACACATTAAGGATTGACAGATTGAGAGCTCTTTCTTGATTATGTGGGTGGTGGTGCATGGCCGTTCTTAGTTGGTGGAGTGATTTGTCTGCTTAATTGCGATAACGAACGAGACCTTAACCTGCTAAATAGCCCGGTCCGCTTTGGCGGGCCGCTGGCTTCTTAGAGGGACTATCGGCTCAAGCCGATGGAAGTTTGAGGCAATAACAGGTCTGTGATGCCCTTAGATGTTCTGGGCCGCACGCGCGCTACACTGACAGAGCCAACGAGTAAATCACCTTGGCCGGAAGGTCTGGGTAATCTTGTTAAACTCTGTCGTGCTGGGGATAGAGCATTGCAATTATTGCTCTTCAACGAGGAATTCCTAGTAAGCGCAAGTCATCAGCTTGCGCTGATTACGTCCCTGCCCTTTGTACACACCGCCCGTCGCTACTACCGATTGAATGGCTCAGTGAGGCCTTCGGACTGGCACAGGGACGTTGGCAACGACGACCCAGTGCCGGAAAGTTGGTCAAACTTGGTCATTTAGAGGAAGTAAAAGTCGTAACAAGGTTTCCGTAGGTGAACCTGCGGAAGGATCATTA
(SEQ ID NO: 26) Symbiotaphrina kochii , control strain CBS 589.63 Grinder beetles Lasioderma serricorne Mycetoma TACCTGGTTGATTCTGCCAGTAGTCATATGCTTGTCTCAAAGATTAAGCCATGCAAGTCTAAGTATAAGCAATCTATACGGTGAAACTGCGAATGGCTCATTAAATCAGTTATCGTTTATTTGATAGTACCTTACTACATGGATAACCGTGGTAATTCTAGAGCTAATACATGCTAAAAACCTCGACTTCGGAAGGGGTGTATTTATTAGATAAAAAACCAATGCCCTTCGGGGCTCCTTGGTGATTCATGATAACTTAACGAATCGCATGGCCTTGCGCCGGCGATGGTTCATTCAAATTTCTGCCCTATCAACTTTCGATGGTAGGATAGTGGCCTACCATGGTTTCAACGGGTAACGGGGAATTAGGGTTCGATTCCGGAGAGGGAGCCTGAGAAACGGCTACCACATCCAAGGAAGGCAGCAGGCGCGCAAATTACCCAATCCCGACACGGGGAGGTAGTGACAATAAATACTGATACAGGGCTCTTTTGGGTCTTGTAATTGGAATGAGTACAATTTAAATCCCTTAACGAGGAACAATTGGAGGGCAAGTCTGGTGCCAGCAGCCGCGGTAATTCCAGCTCCAATAGCGTATATTAAAGTTGTTGCAGTTAAAAAGCTCGTAGTTGAACCTTGGGCCTGGCTGGCCGGTCCGCCTAACCGCGTGTACTGGTCCGGCCGGGCCTTTCCTTCTGGGGAGCCGCATGCCCTTCACTGGGTGTGTCGGGGAACCAGGACTTTTACTTTGAAAAAATTAGAGTGTTCAAAGCAGGCCTATGCTCGAATACATTAGCATGGAATAATAGAATAGGACGTGTGGTTCTATTTTGTTGGTTTCTAGGACCGCCGTAATGATTAATAGGGATAGTCGGGGGCATCAGTATTCAATTGTCAGAGGTGAAATTCTTGGATTTATTGAAGACTAACTACTGCGAAAGCATTTGCCAAGGATGTTTTCATTAATCAGTGAACGAAAGTTAGGGGATCGAAGACGATC AGATACCGTCGTAGTCTTAACCATAAACTATGCCGACTAGGGATCGGGCGATGTTATTATTTTGACTCGCTCGGCACCTTACGAGAAATCAAAGTCTTTGGGTTCTGGGGGGAGTATGGTCGCAAGGCTGAAACTTAAAGAAATTGACGGAAGGGCACCACCAGGAGTGGAGCCTGCGGCTTAATTTGACTCAACACGGGGAAACTCACCAGGTCCAGACACATTAAGGATTGACAGATTGAGAGCTCTTTCTTGATTATGTGGGTGGTGGTGCATGGCCGTTCTTAGTTGGTGGAGTGATTTGTCTGCTTAATTGCGATAACGAACGAGACCTTAACCTGCTAAATAGCCCGGTCCGCTTTGGCGGGCCGCTGGCTTCTTAGAGGGACTATCGGCTCAAGCCGATGGAAGTTTGAGGCAATAACAGGTCTGTGATGCCCTTAGATGTTCTGGGCCGCACGCGCGCTACACTGACAGAGCCAACGAGTACATCACCTTGGCCGGAAGGTCTGGGTAATCTTGTTAAACTCTGTCGTGCTGGGGATAGAGCATTGCAATTATTGCTCTTCAACGAGGAATTCCTAGTAAGCGCAAGTCATCAGCTTGCGCTGATTACGTCCCTGCCCTTTGTACACACCGCCCGTCGCTACTACCGATTGAATGGCTCAGTGAGGCCTTCGGACTGGCACAGGGACGTTGGCAACGACGACCCAGTGCCGGAAAGTTCGTCAAACTTGGTCATTTAGAGGAAGNNNAAGTCGTAACAAGGTTTCCGTAGGTGAACCTGCGGAAGGATCATTA
(SEQ ID NO: 27) Primary extracellular symbiont Master Location 16S rRNA Bacteria that break down fenitrothion Burkholderia sp. SFA1 Riptortus pedestris Intestine
(SEQ ID NO: 28) Burkholderia sp. KM-A Riptortus pedestris Intestine GCAACCCTGGTGGCGAGTGGCGAACGGGTGAGTAATACATCGGAACGTGTCCTGTAGTGGGGGATAGCCCGGCGAAAGCCGGATTAATACCGCATACGATCTACGGAAGAAAGCGGGGGATCCTTCGGGACCTCGCGCTATAGGGGCGGCCGATGGCAGATTAGCTAGTTGGTGGGGTAAAGGCCTACCAAGGCGACGATCTGTAGCTGGTCTGAGAGGACGACCAGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTGGGGAATTTTGGACAATGGGGGCAACCCTGATCCAGCAATGCCGCGTGTGTGAAGAAGGCCTTCGGGTTGTAAAGCACTTTTGTCCGGAAAGAAAACGTCTTGGTTAATACCTGAGGCGGATGACGGTACCGGAAGAATAAGCACCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGTGCGAGCGTTAATCGGAATTACTGGGCGTAAAGCGTGCGCAGGCGGTCTGTTAAGACCGATGTGAAATCCCCGGGCTTAACCTGGGAACTGCATTGGTGACTGGCAGGCTTTGAGTGTGGCAGAGGGGGGTAGAATTCCACGTGTAGCAGTGAAATGCGTAGAGATGTGGAGGAATACCGATGGCGAAGGCAGCCCCCTGGGCCAACACTGACGCTCATGCACGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCCTAAACGATGTCAACTAGTTGTTGGGGATTCATTTCCTTAGTAACGTAGCTAACGCGTGAAGTTGACCGCCTGGGGAGTACGGTCGCAAGATTAAAACTCAAAGGAATTGACGGGGACCCGCACAAGCGGTGGATGATGTGGATTAATTCGATGCAACGCGAAAAACCTTACCTACCCTTGACATGGTCGGAAGTCTGCTGAGAGGTGGACGTGCTCGAAAGAGAACCGGCGCACAGGTGCTGCATGGCTGTCGTCAGCTCGTGTCGTGAG ATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGTCCTTAGTTGCTACGCAAGAGCACTCTAAGGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAGTCCTCATGGCCCTTATGGGTAGGGCTTCACACGTCATACAATGGTCGGAACAGAGGGTTGCCAAGCCGCGAGGTGGAGCCAATCCCAGAAAACCGATCGTAGTCCGGATCGCAGTCTGCAACTCGACTGCGTGAAGCTGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGTCTTGTACACACCGCCCGTCACACCATGGGAGTGGGTTTCACCAGAAGTAGGTAGCCTAACCGCAAGGAGGGCGCTTACCACGGTGGGATTCATGACTGGGGTGAAGT
(SEQ ID NO: 29) Burkholderia sp. KM-G Riptortus pedestris Intestine
(SEQ ID NO: 30) bees Snodgrassella alvi Honey bee ( Apis mellifera ) and Bombus spp. Ileum GAGAGTTTGATCCTGGCTCAGATTGAACGCTGGCGGCATGCCTTACACATGCAAGTCGAACGGCAGCACGGAGAGCTTGCTCTCTGGTGGCGAGTGGCGAACGGGTGAGTAATGCATCGGAACGTACCGAGTAATGGGGGATAACTGTCCGAAAGGATGGCTAATACCGCATACGCCCTGAGGGGGAAAGCGGGGGATCGAAAGACCTCGCGTTATTTGAGCGGCCGATGTTGGATTAGCTAGTTGGTGGGGTAAAGGCCTACCAAGGCGACGATCCATAGCGGGTCTGAGAGGATGATCCGCCACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTGGGGAATTTTGGACAATGGGGGGAACCCTGATCCAGCCATGCCGCGTGTCTGAAGAAGGCCTTCGGGTTGTAAAGGACTTTTGTTAGGGAAGAAAAGCCGGGTGTTAATACCATCTGGTGCTGACGGTACCTAAAGAATAAGCACCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGTGCGAGCGTTAATCGGAATTACTGGGCGTAAAGCGAGCGCAGACGGTTAATTAAGTCAGATGTGAAATCCCCGAGCTCAACTTGGGACGTGCATTTGAAACTGGTTAACTAGAGTGTGTCAGAGGGAGGTAGAATTCCACGTGTAGCAGTGAAATGCGTAGAGATGTGGAGGAATACCGATGGCGAAGGCAGCCTCCTGGGATAACACTGACGTTCATGCTCGAAAGCGTGGGTAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCCTAAACGATGACAATTAGCTGTTGGGACACTAGATGTCTTAGTAGCGAAGCTAACGCGTGAAATTGTCCGCCTGGGGAGTACGGTCGCAAGATTAAAACTCAAAGGAATTGACGGGGACCCGCACAAGCGGTGGATGATGTGGATTAATTCGATGCAACGCGAAGAACCTTACCTGGTCTTGACATGTACGGAATCT CTTAGAGATAGGAGAGTGCCTTCGGGAACCGTAACACAGGTGCTGCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGTCATTAGTTGCCATCATTAAGTTGGGCACTCTAATGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAGTCCTCATGGCCCTTATGACCAGGGCTTCACACGTCATACAATGGTCGGTACAGAGGGTAGCGAAGCCGCGAGGTGAAGCCAATCTCAGAAAGCCGATCGTAGTCCGGATTGCACTCTGCAACTCGAGTGCATGAAGTCGGAATCGCTAGTAATCGCAGGTCAGCATACTGCGGTGAATACGTTCCCGGGTCTTGTACACACCGCCCGTCACACCATGGGAGTGGGGGATACCAGAATTGGGTAGACTAACCGCAAGGAGGTCGCTTAACACGGTATGCTTCATGACTGGGGTGAAGTCGTAACAAGGTAGCCGTAG
(SEQ ID NO: 33) Gilliamella apicola Honey bee ( Apis mellifera ) and Bombus spp. Ileum
(SEQ ID NO: 34) Bartonella apis Honey bee ( Apis mellifera ) Intestine
(SEQ ID NO: 35) Parasaccharibacter apium Honey bee ( Apis mellifera ) Intestine
(SEQ ID NO: 36) Lactobacillus kunkeei Honey bee ( Apis mellifera ) Intestine
(SEQ ID NO: 37) Lactobacillus Firm-4 Honey bee ( Apis mellifera ) Intestine ACGAACGCTGGCGGCGTGCCTAATACATGCAAGTCGAGCGCGGGAAGTCAGGGAAGCCTTCGGGTGGAACTGGTGGAACGAGCGGCGGATGGGTGAGTAACACGTAGGTAACCTGCCCTAAAGCGGGGGATACCATCTGGAAACAGGTGCTAATACCGCATAAACCCAGCAGTCACATGAGTGCTGGTTGAAAGACGGCTTCGGCTGTCACTTTAGGATGGACCTGCGGCGTATTAGCTAGTTGGTGGAGTAACGGTTCACCAAGGCAATGATACGTAGCCGACCTGAGAGGGTAATCGGCCACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCACAATGGACGCAAGTCTGATGGAGCAACGCCGCGTGGATGAAGAAGGTCTTCGGATCGTAAAATCCTGTTGTTGAAGAAGAACGGTTGTGAGAGTAACTGCTCATAACGTGACGGTAATCAACCAGAAAGTCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGATTTATTGGGCGTAAAGGGAGCGCAGGCGGTCTTTTAAGTCTGAATGTGAAAGCCCTCAGCTTAACTGAGGAAGAGCATCGGAAACTGAGAGACTTGAGTGCAGAAGAGGAGAGTGGAACTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAAGAACACCAGTGGCGAAGGCGGCTCTCTGGTCTGTTACTGACGCTGAGGCTCGAAAGCATGGGTAGCGAACAGGATTAGATACCCTGGTAGTCCATGCCGTAAACGATGAGTGCTAAGTGTTGGGAGGTTTCCGCCTCTCAGTGCTGCAGCTAACGCATTAAGCACTCCGCCTGGGGAGTACGACCGCAAGGTTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAGGTCTTGACATCTCCTGCAAGC CTAAGAGATTAGGGGTTCCCTTCGGGGACAGGAAGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGTTACTAGTTGCCAGCATTAAGTTGGGCACTCTAGTGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGACGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGATGGTACAATGAGAAGCGAACTCGCGAGGGGAAGCTGATCTCTGAAAACCATTCTCAGTTCGGATTGCAGGCTGCAACTCGCCTGCATGAAGCTGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCC
(SEQ ID NO: 38) Silkworm Enterococcus bombyx mori Intestine
(SEQ ID NO: 39) Delftia bombyx mori Intestine CAGAAAGGAGGTGATCCAGCCGCACCTTCCGATACGGCTACCTTGTTACGACTTCACCCCAGTCACGAACCCCGCCGTGGTAAGCGCCCTCCTTGCGGTTAGGCTACCTACTTCTGGCGAGACCCGCTCCCATGGTGTGACGGGCGGTGTGTACAAGACCCGGGAACGTATTCACCGCGGCATGCTGATCCGCGATTACTAGCGATTCCGACTTCACGCAGTCGAGTTGCAGACTGCGATCCGGACTACGACTGGTTTTATGGGATTAGCTCCCCCTCGCGGGTTGGCAACCCTCTGTACCAGCCATTGTATGACGTGTGTAGCCCCACCTATAAGGGCCATGAGGACTTGACGTCATCCCCACCTTCCTCCGGTTTGTCACCGGCAGTCTCATTAGAGTGCTCAACTGAATGTAGCAACTAATGACAAGGGTTGCGCTCGTTGCGGGACTTAACCCAACATCTCACGACACGAGCTGACGACAGCCATGCAGCACCTGTGTGCAGGTTCTCTTTCGAGCACGAATCCATCTCTGGAAACTTCCTGCCATGTCAAAGGTGGGTAAGGTTTTTCGCGTTGCATCGAATTAAACCACATCATCCACCGCTTGTGCGGGTCCCCGTCAATTCCTTTGAGTTTCAACCTTGCGGCCGTACTCCCCAGGCGGTCAACTTCACGCGTTAGCTTCGTTACTGAGAAAACTAATTCCCAACAACCAGTTGACATCGTTTAGGGCGTGGACTACCAGGGTATCTAATCCTGTTTGCTCCCCACGCTTTCGTGCATGAGCGTCAGTACAGGTCCAGGGGATTGCCTTCGCCATCGGTGTTCCTCCGCATATCTACGCATTTCACTGCTACACGCGGAATTCCATCCCCCTCTACCGTACTCTAGCCATGCAGTCACAAATGCAGTTCCCAGGTTGAGCCCGGGGATTTCACATCTGTCTTACATAACCGCCTGCGCACGCTTTACGCCCAGTAATTCCGATTAACGCTCG CACCCTACGTATTACCGCGGCTGCTGGCACGTAGTTAGCCGGTGCTTATTCTTACGGTACCGTCATGGGCCCCCTGTATTAGAAGGAGCTTTTTCGTTCCGTACAAAAGCAGTTTACAACCCGAAGGCCTTCATCCTGCACGCGGCATTGCTGGATCAGGCTTTCGCCCATTGTCCAAAATTCCCCACTGCTGCCTCCCGTAGGAGTCTGGGCCGTGTCTCAGTCCCAGTGTGGCTGGTCGTCCTCTCAGACCAGCTACAGATCGTCGGCTTGGTAAGCTTTTATCCCACCAACTACCTAATCTGCCATCGGCCGCTCCAATCGCGCGAGGCCCGAAGGGCCCCCGCTTTCATCCTCAGATCGTATGCGGTATTAGCTACTCTTTCGAGTAGTTATCCCCCACGACTGGGCACGTTCCGATGTATTACTCACCCGTTCGCCACTCGTCAGCGTCCGAAGACCTGTTACCGTTCGACTTGCATGTGTAAGGCATGCCGCCAGCGTTCAATCTGAGCCAGGATCAAACTCTACAGTTCGATCT
(SEQ ID NO: 40) Pelomonas bombyx mori Intestine
(SEQ ID NO: 41)

Any number of bacterial species can be targeted using the compositions or methods described herein. For example, in some cases, the modulating agent may target one species of bacteria. In some cases, the modulator may target at least about any of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30, 40, 50, 60, 70, 80 , 90, 100, 150, 200, 250, 500 or more different types of bacteria. In some instances, the modulating agent may target any of about 1 to about 5, about 5 to about 10, about 10 to about 20, about 20 to about 50, about 50 to about 100, about 100 to about 200, about 200 to about 500, about 10 to about 50, about 5 to about 20, or about 10 to about 100 different bacterial species. In some instances, the modulator may target at least about any of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30, 40, 50, 60, 70, 80 , 90, 100 or more types, classes, orders, families or genera of bacteria.

In some cases, the modulating agent may increase the population of one or more bacteria (e.g., symbiotic bacteria) by at least about any of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more in the host compared to the host to which the modulating agent has not been administered. In some instances, the modulating agent may reduce the population of one or more bacteria (e.g., pathogenic or parasitic bacteria) by at least about any of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80 %, 90% or more in the host compared to the host to which the modulating agent has not been administered. In some cases, the modulating agent can destroy the population of bacteria (eg, pathogenic or parasitic bacteria) in the host organism. In some cases, the modulating agent may increase the level of one or more commensal bacteria by at least about any of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or more in the host body and/or reduce the level of one or more pathogenic bacteria by at least about any of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more in the body host compared to a host to which the modulating agent has not been administered.

In some instances, the modulating agent may alter the bacterial diversity and/or bacterial composition of the host. In some instances, the modulating agent may increase the host's bacterial diversity relative to baseline diversity by at least about any of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or more compared to the host organism, to which the modulating agent was not introduced. In some instances, the modulating agent may reduce host bacterial diversity relative to baseline diversity by at least about any one of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or more compared to the host organism, to which the modulating agent was not introduced.

In some cases, the modulating agent may alter the function, activity, growth and/or division of one or more bacterial cells. For example, a modulating agent may alter the expression of one or more genes in a bacterium. In some cases, the modulating agent may alter the function of one or more proteins in the bacterium. In some instances, the modulating agent may alter the function of one or more cellular structures (eg, cell wall, outer or inner membrane) in the bacterium. In some cases, the modulating agent may kill (eg, lyse) the bacterium.

The target bacterium can live in one or more parts of the insect's body. In addition, the target bacterium may be intracellular or extracellular. In some cases, the bacteria reside in one or more parts of the host's gut, including, for example, the foregut, midgut, and/or hindgut. In some cases, the bacteria live as intracellular bacteria in the cell of the insect host. In some cases, the bacteria inhabit the bacteriocyte of the insect host.

Changes in bacterial populations in the host can be determined using any methods known in the art, such as microarray analysis, polymerase chain reaction (PCR), real-time PCR, flow cytometry, fluorescence microscopy, transmission electron microscopy, fluorescence in situ hybridization (eg, FISH), spectrophotometry, matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS), and DNA sequencing. In some cases, a sample from a host treated with a modulating agent is sequenced (eg, by metagenomic sequencing of 16S rRNA or rDNA) to determine the microbiota of the host after delivery or administration of the modulating agent. In some cases, a sample from a host that has not received the modulating agent is also sequenced to provide a reference.

Mushrooms

Illustrative fungi that can be targeted according to the methods and compositions provided herein include, without limitation , Amylostereum areolatum, Epichloe spp, Pichia pinus, Hansenula capsulate, Daldinia decipien, Ceratocytis spp, Ophiostoma spp, and Attamyces bromatificus. Non-limiting examples of yeast and yeast-like symbionts found in insects include Candida, Metschnikowia, Debaryomyces, Scheffersomyces shehatae and Scheffersomyces stipites, Starmerella, Pichia, Trichosporon, Cryptococcus, Pseudozyma, and yeast-like symbionts from the subtype Pezizomycotina (e.g., Symbiotaphrina bucaphrina and Symbiotaphrina bucaphrina kochii ). Non-limiting examples of yeasts that can be targeted using the methods and compositions of this document are shown in Table 2.

table 2

type of insect Order: family Location of yeast fungi (species) Stegobium paniceum Coleoptera: Anobiidae Mycetoma (= Sitodrepa panicea) ( Saccharomyces ) Caecum ( Torulopsis buchnerii ) Mycetoma between the anterior and midgut Mycetomas ( Symbiotaphrina buchnerii ) Mycetomas and alimentary canal ( Torulopsis buchnerii ) Caecum (Symbiotaphrina buchnerii ) Lasioderma serricorne Coleoptera: Anobiidae Mycetoma between the anterior and midgut ( Symbiotaphrina kochii ) Ernobius abietis Coleoptera: Anobiidae Mycetomas ( Torulopsis karawaiewii ) ( Candida karawaiewii ) Ernobius mollis Coleoptera: Anobiidae Mycetomas ( Torulopsis ernobii ) ( Candida ernobii ) Hemicoelus gibbicollis Coleoptera: Anobiidae Mycetoma larvae Xestobium plumbeum Coleoptera: Anobiidae Mycetomas ( Torulopsis xestobii ) ( Candida xestobii ) Criocephalus rusticus Coleoptera: Cerambycidae Mycetoma Phoracantha semipunctata Coleoptera: Cerambycidae Alimentary canal ( Candida guilliermondii , C. tenuis ) Cecum near midgut ( Candida guilliermondii ) Harpium inquisitor Coleoptera: Cerambycidae Mycetomas ( Candida rhagii ) Harpium mordax
H. sycophanta Coleoptera: Cerambycidae Caecum near midgut ( Candida tenuis ) Gaurotes virginea Coleoptera: Cerambycidae Cecum near midgut ( Candida rhagii ) Leptura rubra Coleoptera: Cerambycidae Caecum near midgut ( Candida tenuis ) Cecum near midgut ( Candida parapsilosis ) Leptura maculicornis Coleoptera: Cerambycidae Cecum near midgut ( Candida parapsilosis ) L. cerambyciformis Leptura sanguinolenta Coleoptera: Cerambycidae Cecum near midgut ( Candida sp. ) Rhagium bifasciatum Coleoptera: Cerambycidae Caecum near midgut ( Candida tenuis ) Rhagium inquisitor Coleoptera: Cerambycidae Cecum near midgut ( Candida guilliermondii ) Rhagium mordax Coleoptera: Cerambycidae Cecum near midgut ( Candida ) Carpophilus hemipterus Coleoptera: Nitidulidae Intestinal tract (10 yeast species) Odontotaenius disjunctus Coleoptera: Passalidae Hindgut ( Entoramus dimorphus ) Odontotaenius disjunctus Coleoptera: Passalidae Gut ( Pichia stipitis , P. segobiensis , Candida shehatae ) Verres sternbergianus ( C. ergatensis ) Scarabaeus semipunctatus Coleoptera: Scarabaeidae Digestive tract (10 yeast species) Chironitis furcifer unknown species Coleoptera: Scarabaeidae Gut ( Trichosporon cutaneum ) Dendroctonus and Ips spp. Coleoptera: Scolytidae Alimentary canal (13 types of yeast) Dendroctonus frontalis Coleoptera: Scolytidae Midgut ( Candida sp. ) Ips sexdentatus Coleoptera: Scolytidae Digestive tract ( Pichia bovis , P. rhodanensis ) Hansenula holstii ( Candida rhagii ) digestive tract ( Candida pulcherina ) Ips typographus Coleoptera: Scolytidae alimentary canal Digestive tract ( Hansenula capsulata , Candida parapsilosis ) Intestine and homogenates of beetles ( Hansenula holstii , H. capsulata , Candida diddensii , C. mohschtana , C. nitratophila , Cryptococcus albidus , C. laurentii ) Trypodendron lineatum Coleoptera: Scolytidae Undefined Xyloterinus politus Coleoptera: Scolytidae Head, thorax, abdomen ( Candida , Pichia , Saccharomycopsis ) Periplaneta americana Dictyoptera: Blattidae Hemocoel ( Candida sp. nov. ) Blatta orientalis Dictyoptera: Blattidae Intestinal tract ( Kluyveromyces blattae ) Blatella germanica Dictyoptera: Blattellidae Hemocel Cryptocercus punctulatus Dictyoptera: Cryptocercidae Hindgut (1 type of yeast) Philophylla heraclei Diptera: Tephritidae Hemocel Aedes (4 species) Diptera: Culicidae Internal microflora (9 genera of yeast fungi) Drosophila pseudoobscura Diptera: Drosophilidae Alimentary canal (24 yeast species) Drosophila (5 species) Diptera: Drosophilidae Goiter (42 types of yeast fungi) Drosophila melanogaster Diptera: Drosophilidae Goiter (8 types of yeast fungi) Drosophila (4 species) Diptera: Drosophilidae Goiter (7 types of yeast fungi) Drosophila (6 species) Diptera: Drosophilidae Gut of larvae (17 types of yeast mushrooms) Drosophila (20 species) Diptera: Drosophilidae Goiter (20 types of yeast fungi) Drosophila (8 species groups) Diptera: Drosophilidae Goiter ( Kloeckera, Candida, Kluyveromyces ) Drosophila serido Diptera: Drosophilidae Goiter (18 types of yeast fungi) Drosophila (6 species) Diptera: Drosophilidae Intestinal epithelium ( Coccidiascus legeri ) Protaxymia melanoptera Diptera Not known ( Candida , Cryptococcus , Sporoblomyces ) Astegopteryx styraci Homoptera: Aphididae Hemocoel and fat body Tuberaphis sp. Homoptera: Aphididae tissue sections Hamiltonaphis styraci Glyphinaphis bambusae Cerataphis sp. Hamiltonaphis styraci Homoptera: Aphididae Hemocoel of the abdomen Cofana unimaculata Homoptera: Cicadellidae fat body Leofa unicolor Homoptera: Cicadellidae fat body Lecaniines etc. Homoptera: Coccoidea Hemolymph, adipose tissue, etc. Lecanium sp. Homoptera: Coccidae Hemolymph, adipose tissue Ceroplastes (4 species) Homoptera: Coccidae blood smears Laodelphax striatellus Homoptera: Delphacidae fat body Eggs Eggs ( Candida ) Nilaparvata lugens Homoptera: Delphacidae fat body Eggs (2 unidentified yeast species) Eggs, nymphs ( Candida ) Eggs (7 unidentified yeast species) Eggs ( Candida ) Nisia nervosa Homoptera: Delphacidae fat body Nisia grandiceps Perkinsiella spp. Sardia rostrata Sogatella furcifera Sogatodes orizicola Homoptera: Delphacidae fat body Amrasca devastans Homoptera: Jassidae Eggs, mycetomas, hemolymph Tachardina lobata Homoptera: Kerriidae Blood smears ( Torulopsis ) Laccifer (=Lakshadia) sp. Homoptera: Kerriidae Blood smears ( Torula variabilis ) Comperia merceti Hymenoptera: Encyrtidae Hemolymph, gut, venom gland Solenopsis invicta Hymenoptera: Formicidae Hemolymph ( Myrmecomyces annellisae ) S. quinquecuspis Solenopsis invicta Hymenoptera: Formicidae Fourth instar larvae ( Candida parapsilosis , Yarrowia lipolytica ) Gut and hemolymph ( Candida parapsilosis , C. lipolytica , C. guillermondii , C. rugosa , Debaryomyces hansenii ) Apis mellifera Hymenoptera: Apidae Digestive tract ( Torulopsis sp. ) Intestinal tract ( Torulopsis apicola ) Digestive tract (8 types of yeast) Intestinal contents (12 yeast species) Intestinal contents (7 types of yeast) Intestines (14 types of yeast mushrooms) Intestinal tract ( Pichia melissophila ) Intestinal tract (7 types of yeast) Alimentary canal ( Hansenula silvicola ) Goiter and intestine (13 types of yeast fungi) Apis mellifera Hymenoptera: Apidae Midgut (9 yeast genera) Anthophora occidentalis Hymenoptera: Anthophoridae Nomia melanderi Hymenoptera: Halictidae Halictus rubicundus Hymenoptera: Halictidae Megachile rotundata Hymenoptera: Megachilidae

Any number of fungal species can be targeted using the compositions or methods described herein. For example, in some cases, the modulating agent may target one type of fungus. In some cases, the modulator may target at least about any of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30, 40, 50, 60, 70, 80 , 90, 100, 150, 200, 250, 500 or more different types of mushrooms. In some instances, the modulating agent may target any of about 1 to about 5, about 5 to about 10, about 10 to about 20, about 20 to about 50, about 50 to about 100, about 100 to about 200, about 200 to about 500, about 10 to about 50, about 5 to about 20, or about 10 to about 100 different mushroom species. In some instances, the modulator may target at least about any of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30, 40, 50, 60, 70, 80 , 90, 100 or more types, classes, orders, families or genera of fungi.

In some cases, the modulating agent may increase the population of one or more fungi (e.g., symbiotic fungi) by at least about any of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more in the host compared to the host to which the modulating agent has not been administered. In some cases, the modulating agent may reduce the population of one or more fungi (e.g., pathogenic or parasitic fungi) by at least about any of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80 %, 90% or more in the host compared to the host to which the modulating agent has not been administered. In some cases, the modulating agent can destroy the population of fungi (eg, pathogenic or parasitic fungi) in the host organism. In some cases, the modulating agent may increase the level of one or more commensal fungi by at least about any of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or more in host organism and/or may reduce the level of one or more pathogenic fungi by at least approximately any of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more in the host organism in comparison with the host organism, to which the modulating agent was not introduced.

In some instances, the modulating agent may alter the fungal diversity and/or fungal composition of the host. In some instances, the modulating agent may increase fungal diversity in the host relative to baseline diversity by at least about any of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or more compared to the host organism, to which the modulating agent was not introduced. In some instances, the modulating agent may reduce the diversity of fungi in the host relative to baseline diversity by at least about any one of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or more compared to the host organism, to which the modulating agent was not introduced.

In some cases, the modulating agent may alter the function, activity, growth and/or division of one or more fungi. For example, a modulating agent may alter the expression of one or more genes in a fungus. In some cases, the modulator may alter the function of one or more proteins in the fungus. In some cases, the modulating agent may alter the function of one or more cellular components in the fungus. In some cases, the modulating agent can kill the fungus.

In addition, the target fungus may reside in one or more parts of the insect's body. In some cases, the fungus lives in or on one or more parts of the insect's gut, including, for example, the foregut, midgut, and/or hindgut. In some cases, the fungus lives extracellularly in the hemolymph, fat bodies, or in specialized structures in the host organism.

Changes in the population of fungi in the host can be determined using any methods known in the art, such as microarray analysis, polymerase chain reaction (PCR), real-time PCR, flow cytometry, fluorescence microscopy, transmission electron microscopy, fluorescence in situ hybridization (eg, FISH), spectrophotometry, matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS), and DNA sequencing. In some cases, a sample from a host treated with a modulating agent is sequenced (eg, by metagenomic sequencing) to determine the microbiota of the host after delivery or administration of the modulating agent. In some cases, a sample from a host that has not received the modulating agent is also sequenced to provide a reference.

modulating means

The modulator of the methods and compositions provided herein may include a phage, a polypeptide, a small molecule, an antibiotic, a secondary metabolite, a bacterium, a fungus, or any combination thereof.

phage

The modulator described herein may include a phage (eg, lytic phage or non-lytic phage). In some cases, an effective concentration of any phage described herein can alter the level, activity, or metabolism of one or more microorganisms (described herein) living in the host described herein, the change resulting in increased host fitness. In some instances, the modulating agent includes at least one phage selected from the orders Tectiviridae, Myoviridae, Siphoviridae, Podoviridae, Caudovirales, Lipothrixviridae, Rudiviridae, or Ligamenvirales . In some cases, the composition contains at least one phage selected from the family Myoviridae, Siphoviridae, Podoviridae, Lipothrixviridae, Rudiviridae, Ampullaviridae, Bicaudaviridae, Clavaviridae, Corticoviridae, Cystoviridae, Fuselloviridae, Gluboloviridae, Guttaviridae, Inoviridae, Leviviridae, Microviridae, Plasmaviridae and Tectiviridae. Additional non-limiting examples of phages useful in methods and compositions are shown in Table 3.

Table 3. Examples of phages and target bacteria

Phage and Access No. Target bacterium Host-target SA1(NC_027991), phiP68(NC_004679) Staphylococcus sp. Family Apidae WO (AB036666.1) Wolbachia sp. Aedes aegypti ; Drosophila melanogaster ; Plasmodium sp ; Teleogryllus taiwanemma ; bombyx mori KL1 (NC_018278), BcepNazgul (NC_005091) PhiE125 (NC_003309) Burkholderia sp. Riptortus sp. ; Pyrrhocoris apterus. Fern (NC_028851), Xenia (NC_028837), HB10c2 (NC_028758) Paenibacillus larvae Bumblebees: Bombus sp. ; honey bees: A. mellifera CP2 (NC_020205), XP10 (NC_004902), XP15 (NC_007024), phiL7 (NC_012742) Xanthomonas sp. Plebeina denoiti ; family Apidae ; Apis mellifera ; family Drosophilidae and family Chloropidae PP1 (NC_019542), PM1 (NC_023865) Pectobacterium
carotovorum subsp .
carotovorum Family Apidae ΦRSA1 (NC_009382),
ΦRSB1 (NC_011201), ΦRSL1 (NC_010811), RSM1 (NC_008574) Ralstonia solanacearum bombyx mori SF1(NC_028807) Streptomyces scabies Philanthus sp. ; Trachypus sp. ECML-4 (NC_025446), ECML-117 (NC_025441), ECML-134 (NC_025449) Escherichia coli Family Apidae ;
Varroa destructor SSP5(JX274646.1), SSP6(NC_004831), SFP10(NC_016073), F18SE(NC_028698) Salmonella sp. Family Drosophilidae (NC_001416), Bcp1 (NC_024137) Bacillus sp. Gypsy moth; Lymantria dispar ; Varroa destructor Phi1 (NC_009821) Enterococcus sp. Schistocerca gragaria ΦKMV (NC_005045), ΦEL(AJ697969.1), ΦKZ (NC_004629) Pseudomonas sp. Lymantria dispar ; family Apidae A2 (NC_004112), fig1e (NC_004305) Lactobacillus sp. Family Apidae ; family Drosophila ; Varroa destructor KLPN1 (NC_028760) Klebsiella sp. C.capitata vB_AbaM_Acibel004 (NC_025462), vB_AbaP_Acibel007 (NC_025457) Acinetobacter sp. Schistocerca gragaria

In some cases, the modulating agent includes a lytic phage. Thus, upon delivery of a lytic phage to a bacterial cell residing in the host organism, the phage causes lysis in the target bacterial cell. In some cases, a lytic phage purposefully affects a bacterium living in an insect host and destroys it with an increase in the host's fitness. Alternatively or additionally, the modulating agent phage may be non-lytic phage (also referred to as lysogenic or temperate phage). Thus, after delivery of a non-lytic phage to a bacterial cell living in the host organism, the bacterial cell can remain viable and able to stably maintain the expression of genes encoded in the phage genome. In some cases, a non-lytic phage is used to alter gene expression in a bacterium living in an insect host to improve host fitness. In some cases, the modulating agent includes a mixture of lytic and non-lytic phage.

The modulator described herein includes phage with a narrow or broad spectrum of target bacteria. In some cases, the phage has a narrow range of target bacteria. In some cases, the phage is a promiscuous phage with a wide range of target bacteria. For example, a promiscuous phage can target at least about any one of 5, 10, 20, 30, 40, 50 or more bacteria living in the host organism. A phage with a narrow spectrum of target bacteria can target a particular strain of bacteria in the host without adversely affecting another, eg, non-target, bacterium in the host. For example, a phage can target no more than about any one of 50, 40, 30, 20, 10, 8, 6, 4, 2, or 1 bacteria living in the host organism.

The compositions described herein may contain any number of phages, such as at least about any number of 1, 2, 3, 4, 5, 10, 15, 20, 30, 40, 50, 100 or more phages. . In some cases, the composition contains a phage from one or more families (for example, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more phages), one or more orders (for example, 2, 3, 4, 5 , 6, 7, 8, 9, 10 or more phages) or one or more species (e.g. 1, 2, 3, 4, 5, 10, 15, 20, 30, 40, 50, 100 or more phages). Compositions containing one or more phages are also referred to herein as "phage cocktails". Phage cocktails are useful because they target a wider range of bacterial hosts. In addition, they target each strain (i.e., subspecies) of bacteria of numerous orthogonal phages, thereby preventing or significantly delaying the emergence of resistance. In some cases, the cocktail contains several phages that target one bacterial species. In some cases, the cocktail contains several phages that target several bacterial species. In some cases, a single-phage "cocktail" contains one promiscuous phage (ie, a phage with a wide host spectrum) that targets many strains within a species.

Suitable concentrations of phage in the modulator described herein depend on factors such as potency, survival rate, phage tolerance, number of different phages and/or types of lysine in the compositions, composition and methods of application of the composition. In some cases, the phage is in a liquid or solid composition. In some instances, the concentration of each phage in any of the compositions described herein is at least approximately any one ^of 102, ¹⁰³ , ¹⁰⁴ , ¹⁰⁵ , ¹⁰⁶ , ¹⁰⁷ , ¹⁰⁸ , ¹⁰⁹ , ¹⁰¹⁰ or more PFU / ml. In some cases, the concentration of each phage in any of the compositions described herein is no more than approximately any of 10 ² , 10 ³ , 10 ⁴ , 10 ⁵ , 10 ⁶ , 10 ⁷ , 10 ⁸ , 10 ⁹ PFU/ml . In some instances, the concentration of each phage in the composition is any of from about ¹⁰² to about ¹⁰³ , from about ¹⁰³ to about ¹⁰⁴ , from about ¹⁰⁴ to about ¹⁰⁵ , from about ¹⁰⁵ to about ¹⁰⁶ , from about 10 ⁷ to about 10 ⁸ , about 10 ⁸ to about 10 ⁹ , about 10 ² to about 10 ⁴ , about 10 ⁴ to about 10 ⁶ , about 10 ⁶ to about 10 ⁹ , or about 10 ³ to about 10 ⁸ PFU/ml. In some cases, if the composition contains at least two types of phage, then the concentration of each type of phage may be the same or different. For example, in some cases the concentration of one phage in the cocktail is approximately 10 ⁸ PFU/ml and the concentration of the second phage in the cocktail is approximately 10 ⁶ PFU/ml.

A modulator comprising a phage as described herein may be contacted with a target host in an amount and for a time sufficient to: (a) achieve a target level (e.g., a predetermined or threshold level) of phage concentration within host-target; (b) achieving a target level (eg, a predetermined or threshold level) of phage concentration within the gut of the target host; (c) achieving a target level (eg, a predetermined or threshold level) of phage concentration within the target host bacteriocyte; (d) modulating the level or activity of one or more microorganisms (eg, endosymbionts) in the target host or/and (e) modulating the fitness of the target host.

As illustrated in examples 7 and 9, phages can be used as modulators by removing bacterial pathogens such as Serratia marcescens , Erwinia catotovora and Pseudomonas enzomophila from insect hosts such as crickets.

Polypeptides

Numerous polypeptides (eg, bacteriocin, R-type bacteriocin, C-rich nodule peptide, antimicrobial peptide, lysine, or bacteriocyte regulatory peptide) can be used in the compositions and methods described herein. In some cases, an effective concentration of any peptide or polypeptide described herein can alter the level, activity or metabolism of one or more microorganisms (described herein) living in the host, the change resulting in increased host fitness. The polypeptides included herein may include naturally occurring polypeptides or recombinantly produced variants. For example, a polypeptide may be a functionally active variant of any of the polypeptides described herein by at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79 %, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical, for example, in a certain area or in the entire sequence, the sequence of the polypeptide described in this document, or a naturally occurring polypeptide.

A modulator comprising a polypeptide described herein may be contacted with a target host in an amount and for a time sufficient to: (a) achieve a target level (e.g., a predetermined or threshold level) of concentration within the host - targets; (b) reaching a target level (eg, a predetermined or threshold level) of the concentration within the gut of the target host; (c) achieving a target level (eg, a predetermined or threshold level) of the concentration within the target host bacteriocyte; (d) modulating the level or activity of one or more microorganisms (eg, endosymbionts) in the target host or/and (e) modulating the fitness of the target host.

Bacteriocins

The modulator described herein may include bacteriocin. In some cases, bacteriocin is produced naturally by Gram-positive bacteria such as Pseudomonas, Streptomyces, Bacillus, Staphylococcus, or lactic acid bacteria (LABs such as Lactococcus lactis ). In some cases, bacteriocin is produced naturally by Gram-negative bacteria such as Hafnia alvei , Citrobacter freundii , Klebsiella oxytoca, Klebsiella pneumonia, Enterobacter cloacae , Serratia plymiticum, Xanthomonas campestris, Erwinia carotovora, Ralstonia solanacearum, or Escherichia coli . Illustrative bacteriocins include, without limitation, LAB class I-IV antibiotics (such as lantibiotics), colicins, microcins, and pyocins. Non-limiting examples of bacteriocins are shown in Table 4.

Table 4. Examples of bacteriocins

Class Name Producer targets Subsequence Class I Lowlands Lactococcus lactis Active against Gram-positive bacteria:
Enterococcus - Lactobacillus - Lactococcus - Leuconostoc -
Listeria - clostridium ITSISLCTPGCKTGALMGCNMKTATCHCSIHVSK
(SEQ ID NO: 42) Epidermin Staphylococcus epidermis Gram-positive bacteria IASKFICTPGCAKTGSFNSYCC
(SEQ ID NO: 43) Class II Class IIa Pediocin PA-1 Pediococcus acidilactici Pediococci - Lactobacilli - Leuconostoc - Brochothrix thermosphacta - propionibacteria - Bacilli - Enterococci - Staphylococci - Listeria clostridia - Listeria monocytogenes - Listeria innocua KYYGNGVTCG KHSCSVDWGKATTCIINNGAMAWATGGHQGNHKC
(SEQ ID NO: 44) Class IIb Enterocin P Enterococcus faecium Lactobacillus sakei - Enterococcus faecium ATRSYGNGVYCNNSKCWVNWGEAKENIAGIVISGWASGLAGMGH
(SEQ ID NO: 45) Class IIc Lactococcin G Streptococcus lactis Gram-positive bacteria GTWDDIGQGIGRVAYWVGKAMGNMSDVNQASRINRKKKH
(SEQ ID NO: 46) Class IId Lactacin-F Lactobacillus johnsonii Lactobacilli - Enterococcus faecalis NRWGDTVLSAASGAGTGIKACKSFGPWGMAICGVGGAAIGGYFGYTHN
(SEQ ID NO: 47) Class III Class IIIa Enterocin AS-48 Enterococcus faecalis Broad Spectrum: Gram-positive and Gram-negative bacteria MAKEFGIPAAVAGTVLNVVEAGGWVTTIVSILTAVGSGGLSLLAAAGRESIKAYLKKEIKKKGKRAVIAW
(SEQ ID NO: 48) Class IIIb Aureocin A70 Staphylococcus aureus Broad Spectrum: Gram-positive and Gram-negative bacteria MSWLNFLKYIAKYGKKAVSAAWKYKGKVLEWLNVGPTLEWVWQKLKKIAGL
(SEQ ID NO: 49) Class IV Garvicin A Lactococcus garvieae Broad Spectrum: Gram-positive and Gram-negative bacteria IGGALGNALNGLGTWANMMNGGGFVNQWQVYANKGKINQYRPY
(SEQ ID NO: 50) Unclassified Colicin V Escherichia coli Active against Escherichia coli (as well as closely related bacteria) - Enterobacteriaceae MRTLTLNELDSVSGGASGRDIAMAIGTLSGQFVAGGIGAAAGGVAGGAIYDYASTHKPNPAMSPSGLGGTIKQKPEGIPSEAWNYAAGRLCNWSPNNLSDVCL
(SEQ ID NO: 51)

In some cases, the bacteriocin is a colicin, pyocin or microcin produced by Gram-negative bacteria. In some cases, the bacteriocin is colicin. The colicin can be a group A colicin (eg, uses the Tol system to penetrate the outer membrane of the target bacterium) or a group B colicin (eg, uses the Ton system to penetrate the outer membrane of the target bacterium). In some cases, the bacteriocin is microcin. The microcin can be class I microcin (eg, <5 kDa, has post-translational modifications) or class II microcin (eg, 5-10 kDa, with or without post-translational modifications). In some instances, class II microcin is class IIa microcin (eg, requires more than one gene to synthesize and assemble functional peptides) or class IIb microcin (eg, linear peptides with or without post-translational modifications at the C-terminus). In some cases, the bacteriocin is pyocin. In some cases, pyocin is R-pyocin, F-pyocin, or S-pyocin.

In some cases, the bacteriocin is a class I, class II, class III, or class IV bacteriocin produced by Gram-positive bacteria. In some cases, the modulating agent includes a class I bacteriocin (eg, antibiotics containing lanthionine (lantibiotics) produced by Gram-positive bacteria). Class I bacteriocins or lantibiotics may be a low molecular weight peptide (eg, less than about 5 kDa) and may have amino acid residues with post-translational modifications (eg, lanthionine, β-methyllanthionine, or dehydrogenated amino acids).

In some cases, the bacteriocin is a class II bacteriocin (eg, non-lantibiotic compounds produced by Gram-positive bacteria). Many of these are positively charged lanthionine-free peptides that, unlike lantibiotics, do not undergo extensive post-translational modification. Class II bacteriocin may belong to one of the following subclasses: "pediocin-like" bacteriocins (eg, pediocin PA-1 and carnobacteriocin X (class IIa)); bipeptide bacteriocins (eg lactacin F and ABP-118 (class IIb)); circular bacteriocins (eg, carnocycline A and enterocin AS-48 (class IIc)) or unmodified linear non-pediocin-like bacteriocins (eg, epidermycin NI01 and lactococcin A (class IId)).

In some cases, the bacteriocin is a class III bacteriocin (eg, produced by Gram-positive bacteria). Class III bacteriocins may have a molecular weight greater than 10 kDa and may be thermally unstable proteins. Class III bacteriocins can be further subdivided into group IIIA and group IIIB bacteriocins. Group IIIA bacteriocins include bacteriolytic enzymes that kill sensitive strains through cell wall lysis, such as enterolysin A. Group IIIB bacteriocins include non-lytic proteins such as caseicin 80, helveticin J, and lactacin B.

In some cases, the bacteriocin is a class IV bacteriocin (eg, produced by Gram-positive bacteria). Class IV bacteriocins are a group of complex proteins associated with other lipid or carbohydrate moieties that appear to be required for activity. They are relatively hydrophobic and thermostable. Examples of class IV bacteriocins are leukocin S, lactocin 27, and lactocin S.

In some cases, the bacteriocin is an R-type bacteriocin. R-type bacteriocins are contractile bactericidal protein complexes. Some R-type bacteriocins have a contractile structure similar to a phage outgrowth. The C-terminal region of the phage outgrowth filament protein determines the specificity of the binding to the target. They can attach to target cells via a receptor-binding protein, such as a filament of a process. Attachment is followed by contraction of the outer sheath and insertion of the core through the shell of the target bacterium. Penetration of the core leads to rapid depolarization of the cell membrane potential and instantaneous cell death. Contact with a single particle of R-type bacteriocin can lead to cell death. An R-type bacteriocin, for example, may be heat-labile, moderately acid-resistant, trypsin-resistant, centrifugally sedimentable, electron microscopy-detectable, or a combination of these properties. Other R-type bacteriocins may be complex molecules, including numerous proteins, polypeptides, or subunits, and may resemble the outgrowth structure of bacteriophages of the Myoviridae family. In naturally occurring R-type bacteriocins, subunit structures can be encoded by the bacterial genome, such as that of C. difficile or P. aeruginosa , and form R-type bacteriocins that act as a natural defense against other bacteria. In some cases, the R-type bacteriocin is pyocin. In some cases, pyocin is R-pyocin, F-pyocin, or S-pyocin.

In some instances, the bacteriocin is a functionally active variant of the bacteriocins described herein. In some cases, the bacteriocin variant is at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83 %, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical , for example, in a certain area or in the entire sequence, the sequence of the bacteriocin described herein, or the naturally occurring bacteriocin.

In some instances, bacteriocin can be bioengineered according to standard methods to modulate its biological activity, eg increase or decrease or regulate it, or to determine its target microorganisms. In other cases, bacteriocin is produced by the translation apparatus (eg, ribosome, etc.) of the microbial cell. In some cases, bacteriocin is synthesized chemically. Some bacteriocins can be formed from a polypeptide precursor. The polypeptide precursor may be cleaved (eg, processed by a protease) to form the bacteriocin polypeptide itself. Thus, in some cases, bacteriocin is produced from a precursor polypeptide. In some other cases, bacteriocin includes a polypeptide that has undergone post-translational modifications, such as cleavage or the addition of one or more functional groups.

The bacteriocins described herein may be formulated for any of the uses described herein. The compositions disclosed herein may contain any number or type (e.g., classes) of bacteriocins, such as at least about any of 1 bacteriocin, 2, 3, 4, 5, 10, 15, 20, 30, 40, 50, 100 or more bacteriocins. Suitable concentrations of each bacteriocin in the compositions described herein depend on such factors as the potency, stability of the bacteriocin, the amount of different types of bacteriocin in the compositions, composition and methods of application of the composition. In some instances, the concentration of each bacteriocin in the liquid composition is from about 0.01 ng/mL to about 100 mg/mL. In some instances, the concentration of each bacteriocin in the solid composition is from about 0.01 ng/g to about 100 mg/g. In some cases, if the composition contains at least two types of bacteriocins, then the concentration of each type of bacteriocins may be the same or different. In some instances, the bacteriocin is provided in a composition containing a bacterial cell that secretes the bacteriocin. In some instances, the bacteriocin is provided in a composition containing a polypeptide (eg, a polypeptide isolated from a bacterial cell).

Bacteriocins can neutralize (eg, destroy) at least one microorganism other than the individual bacterial cell in which the polypeptide is formed, including cells clonally related to the bacterial cell and other microbial cells. Thus, a bacterial cell can exhibit cytotoxic or growth-inhibiting effects against a variety of microorganisms through the secretion of bacteriocins. In some cases, bacteriocin targets one or more species of bacteria living in the host organism and destroys them through the formation of pores in the cytoplasmic membrane, cell wall destruction (for example, peptidoglycanase activity), or nuclease activity (for example, DNase activity, 16S rRNase activity, or tRNase activity).

In some cases, bacteriocin is characterized by neutralizing activity. The neutralizing activity of bacteriocins may include, without limitation, microbial arrest or cytotoxicity. Some bacteriocins are characterized by cytotoxic activity, and thus can kill microorganisms, such as bacteria, yeasts, algae, etc. Some bacteriocins can inhibit the multiplication of microorganisms, such as bacteria, yeasts, algae, etc., for example by stopping the cell cycle.

In some cases, bacteriocin is characterized by destroying activity. The killing mechanism of bacteriocins is specific to each group of bacteriocins. In some cases, bacteriocin is characterized by a narrow spectrum of biological activity. Bacteriocins are known for their very high activity against their target strains. The activity of some bacteriocins is limited to strains that are closely related to the bacteriocin-producing strain (narrow spectrum biological activity). In some cases, bacteriocin is characterized by a broad spectrum of biological activity in relation to a wide range of genera.

In some cases, bacteriocins interact with a receptor molecule or docking molecule on the cell membrane of the target bacterium. For example, nisin is highly active against its target bacterial strains, demonstrating antimicrobial activity even at a single nanomolar concentration. The nisin molecule has been shown to bind to lipid II, which is the main carrier of peptidoglycan subunits from the cytoplasm to the cell wall.

In some cases, bacteriocin is characterized by antifungal activity. A number of bacteriocins with anti-yeast or anti-fungal activity have been identified. For example, bacteriocins from Bacillus have been shown to have neutralizing activity against certain yeast strains (see, for example, Adetunji and Olaoye, Malaysian Journal of Microbiology 9:130-13, 2013). In another example, an Enterococcus faecalis peptide has been shown to have neutralizing activity against Candida species (see, for example, Shekh and Roy, BMC Microbiology 12:132, 2012). In another example, bacteriocins from Pseudomonas have been shown to have neutralizing activity against fungi such as Curvularia lunata , Fusarium spp., Helminthosporium spp., and Biopolaris spp . (See, e.g., Shalani and Srivastava, The Internet Journal of Microbiology Volume 5 Number 2, 2008). In another example, botrycidin AJ1316 and alirin B1 from B. subtilis were shown to have antifungal activity.

A modulating agent comprising a bacteriocin as described herein may be contacted with a target host in an amount and for a period of time sufficient to: host-target; (b) achieving a target level (eg, a predetermined or threshold level) of bacteriocin concentration within the gut of the target host; (c) reaching a target level (eg, a predetermined or threshold level) of bacteriocin concentration within a target host bacteriocyte; (d) modulating the level or activity of one or more microorganisms (eg, endosymbionts) in the target host or/and (e) modulating the fitness of the target host.

Lysines

The modulator described herein may include lysine (eg, also known as murein hydrolase or peptidoglycan autolysin). Any lysine suitable for inhibiting the host bacterium may be used. In some cases, lysine is a lysine that can be produced naturally by a bacterial cell. In some cases, lysine is a lysine that can be produced naturally by a bacteriophage. In some cases, lysine is obtained from a phage that inhibits a bacterium that lives in the host organism. In some cases, lysine is designed based on naturally occurring lysine. In some cases, the lysine is engineered for secretion by the host bacterium, for example, by introducing a signal peptide into the lysine. In some cases, lysine is used in combination with phage choline. In some cases, lysine is expressed by a recombinant bacterial host that is not sensitive to lysine. In some cases, lysine is used to inhibit a gram-positive or gram-negative bacterium living in the host organism.

The lysine may be any class of lysine and may have one or more substrate specificities. For example, the lysine may be a glycosidase, an endopeptidase, a carbopeptidase, or a combination thereof. In some cases, lysine cleaves the β-1-4-glycosidic bond in the sugar fragment of the cell wall, the amide bond connecting the sugar and peptide fragments of the bacterial cell wall, and/or the peptide bonds between the peptide fragments of the cell wall. Lysine may belong to one or more specific lysine groups depending on the cleavage site in the peptidoglycan. In some cases, lysine is N -acetyl-β-D-muramidase (for example, lysozyme), lytic transglycosylase, N -acetyl-β-D-glucosaminidase, N -acetylmuramyl-L-alanine amidase, L,D-endopeptidase, D, D-endopeptidase, D,D-carboxypeptidase, L,D-carboxypeptidase or L,D-transpeptidase. Non-limiting examples of lysines and forms of their activity are shown in table 5.

Table 5. Examples of lysines

Target bacterium Producer Lysines Activity Subsequence S. pneumoniae Cp1 cpl-1 Muramidase MVKKNDLFVDVSSHNGYDITGILEQMGTTNTIIKISESTTYLNPCLSAQVEQSNPIGFYHFARFGGDVAEAEREAQFFLDNVPMQVKYLVLDYEDDPSGDAQANTNACLRFMQMIADAGYKPIYYSYKPFTHDNVDYQQILAQFPNSLWIAGYGLNDGTANFEYFPSMDGIRWWQYSSNPFDKNIVLLDDEEDDKPKTAGTWKQDSKGWWFRRNNGSFPYNKWEKIGGVWYYFDSKGYCLTSEWLKDNEKWYYLKDNGAMATGWVLVGSEWYYMDDSGAMVTGWVKYKNNWYYMTNERGNMVSNEFIKSGKGWYFMNTNGELADNPSFTKEPDGLITVA
(SEQ ID NO: 52) S. pneumoniae Dp-1 Pal Amidase MGVDIEKGVAWMQARKGRVSYSMDFRDGPDSYDCSSSMYYALRSAGASSAGWAVNTEYMHAWLIENGYELISENAPWDAKRGDIFIWGRKGASAGAGGHTGMFIDSDNIIHCNYAYDGISVNDHDERWYYAGQPYYYVYRLTNANAQPAEKKLGWQKDATGFWYARANGTYPKDEFEYIEENKSWFYFDDQGYMLAEKWLKHTDGNWYWFDRDGYMATSWKRIGESWYYFNRDGSMVTGWIKYYDNWYYCDATNGDMKSNAFIRYNDGWYLLLPDGRLADKPQFTVEPDGLITAKV
(SEQ ID NO: 53) S. pyogenes C1 C1 Amidase But. B. anthracis PlayG Amidase MEIQKKLVDPSKYGTKCPYTMKPKYITVHNTYNDAPAENEVSYMISNNNEVSFHIAVDDKKAIQGIPLERNAWACGDGNGSGNRQSISVEICYSKSGGDRYYKAEDNAVDVVRQLMSMYNIPIENVRTHQSWSGKYCPHRMLAEGRWGAFIQKVKNGNVATTSPTKQNIIQSGAFSPYETPDVMGALTSLKMTADFILTSDAGLTYK
(SEQ ID NO: 54) B. anthracis Ames Prophage PlyPH Amidase But. E. faecalis and E. faecium Phi1 PlyV12 Amidase But. S. aureus ΦMR11 MV-L Endopeptidase and amidase MQAKLTKKEFIEWLKTSEGKQFNVDLWYGFQCFDYANAGWKVLFGLLLKGLGAKDIPFANNFDGLATVYQNTPDFLAQPGDMVVFGSNYGAGYGHVAWVIEATLDYIIVYEQNWLGGGWTDRIEQPGWGWEKVTRRQHAYDFPMWFIRPNFKSETAPRSIQSPTQASKKETAKPQPKAVELKIIKDVVKGYDLPKRGGNPKGIVIHNDAGSKGATAEAYRNGLVNAPLSRLEAGIAHSYVSGNTVWQALDESQVGWHTANQLGNKYYYGIEVCQSMGADNATFLKNEQATFQECARLLKKWGLPANRNTIRLHNEFTSTSCPHRSSVLHTGFDPVTRGLLPEDKQLQLKDYFIKQIRVYMDGKIPVATVSNESSASSNTVKPVASAWKRNKYGTYYMEENARFTNGNQPITVRKIGPFLSCPVAYQFQPGGYCDYTEVMLQDGHVWVGYTWEGQRYYLPIRTWNGSAPPNQILGDLWGEIS
(SEQ ID NO: 55) S. pyogenes C1 plyc Amidase But. S. agalactiae B30 Lysine GBS Muramidase and endopeptidase MVINIEQAIAWMASRKGKVTYSMDYRNGPSSYDCSSSVYFALRSAGASDNGWAVNTEYEHDWLIKNGYVLIAENTNWNAQRGDIFIWGKRGASAGAFGHTGMFVDPDNIIHCNYGYNSITVNNHDEIWGYNGQPYVYAYRYSGKQSNAKVDNKSVVSKFEKELDVNTPLSNSNMPYYEATISEDYYVEIGARSNQPDVNSTDKELLVAGWAG
(SEQ ID NO: 56) S. aureus P68 Lys16 Endopeptidase But. S. aureus K LysK Amidase and endopeptidase MAKTQAEINKRLDAYAKGTVDSPYRVKKATSYDPSFGVMEAGAIDADGYYHAQCQDLITDYVLWLTDNKVRTWGNAKDQIKQSYGTGFKIHENKPSTVPKKGWIAVFTSGSYEQWGHIGIVYDGGNTSTFTILEQNWNGYANKKPTKRVDNYYGLTHFIEIPVKAGTTVKKETAKKSASKTPAPKKKATLKVSKNHINYTMDKRGKKPEGMVIHNDAGRSSGQQYENSLANAGYARYANGIAHYYGSEGYVWEAIDAKNQIAWHTGDGTGANSGNFRFAGIEVCQSMSASDAQFLKNEQAVFQFTAEKFKEWGLTPNRKTVRLHMEFVPTACPHRSMVLHTGFNPVTQGRPSQAIMNKLKDYFIKQIKNYMDKGTSSSTVVKDGKTSSASTPATRPVTGSWKKNQYGTWYKPENATFVNGNQPIVTRIGSPFLNAPVGGNLPAGATIVYDEVCIQAGHIWIGYNAYNGNRVYCPVRTCQGVPPNQIPGVAWGVFK
(SEQ ID NO: 57) L. monocytogenes A118 Ply118 Amidase MTSYYYSRSLANVNKLADNTKAAARKLLDWSESNGIEVLIYETIRTKEQQAANVNSGASQTMRSYHLVGQALDFVMAKGKTVDWGAYRSDKGKKFVAKAKSLGFEWGGDWSGFVDNPHLQFNYKGYGTDTFGKGASTSNSSKPSADTNTNSLGLVDYMNLNKLDSSFANRKKLATSYGIKNYSGTATQNTTLLAKLKAGKPHTPASKNTYYTENPRKVKTLVQCDLYKSVDFTTKNQTGGTFPPGTVFTISGMGKTKGGTPRLKTKSGYYLTANTKFVKKI
(SEQ ID NO: 58) L. monocytogenes A511 Ply511 Amidase MVKYTVENKIIAGLPKGKLKGANFVIAHETANSKSTIDNEVSYMTRNWKNAFVTHFVGGGGRVVQVANVNYVSWGAGQYANSYSYAQVELCRTSNATTFKKDYEVYCQLLVDLAKKAGIPITLDSGSKTSDKGIKSHKWVADKLGGTTHQDPYAYLSSWGISKAQFASDLAKVSGGGNTGTAPAKPSTPAPKPSTPSTNLDKLGLVDYMNAKKMDSSYSNRDKLAKQYGIANYSGTASQNTTLLSKIKGGAPKPSTPAPKPSTSTAKKIYFPPNKGNWSVYPTNKAPVKANAIGAINPTKFGGLTYTIQKDRGNGVYEIQTDQFGRVQVYGAPSTGAVIKK
(SEQ ID NO: 59) L. monocytogenes A500 Ply500 Endopeptidase MALTEAWLIEKANRKLNAGGMYKITSDKTRNVIKKMAKEGIYLCVAQGYRSTAEQNALYAQGRTKPGAIVTNAKGGQSNHNYGVAVDLCLYTNDGKDVIWESTTSRWKKVVAAMKAEGFKWGGDWKSFKDYPHFELCDAVSGEKIPAATQNTNTNSNRYEGKVIDSAPLLPKMDFKSSPFRMYKVGTEFLVYDHNQYWYKTYIDDKLYYMYKSFCDVVAKKDAKGRIKVRIKSAKDLRIPVWNNIKLNSGKIKWYAPNVKLAWYNYRRGYLELWYPNDGWYYTAEYFLK
(SEQ ID NO: 60) S. pneumoniae ΦDp-1 Pal, S Endopeptidase and amidase But. S. agalactiae Prophage LambdaSa1 LambdaSa1 glycosidase MVINIEQAIAWMASRKGKVTYSMDYRNGPSSYDCSSSVYFALRSAGASDNGWAVNTEYEHDWLIKNGYVLIAENTNWNAQRGDIFIWGKRGASAGAFGHTGMFVDPDNIIHCNYGYNSITVNNHDEIWGYNGQPYVYAYRYARKQSNAKVDNQSVVSKFEKELDVNTPLSNSNMPYYEATISEDYYVESKPDVNSTDKELLVAGY
(SEQ ID NO: 61) S. agalactiae Prophage LambdaSa2 LambdaSa2 glycosidase and endopeptidase MEINTEIAIAWMSARQGKVSYSMDYRDGPNSYDCSSSVYYALRSAGASSAGWAVNTEYMHDWLIKNGYELIAENVDWNAVRGDIAIWGMRGHSSGAGGHVVMFIDPENIIHCNWANNGITVNNYNQTAAASGWMYCYVYRLKSGASTQGKSLDTLVKETLAGNYGNGEARKAVLGNQYEAVMSVINGKTTTNQKTVDQLVQEVIAGKHGNGEARKKSLGSQYDAVQKRVTELLKKQPSEPFKAQEVNKPTETKTSQTELTGQATATKEEGDLSFNGTILKKAVLDKILGNCKKHDILPSYALTILHYEGLWGTSAVGKADNNWGGMTWTGQGNRPSGVTVTQGSARPSNEGGHYMHYASVDDFLTDWFYLLRAGGSYKVSGAKTFSEAIKGMFKVGGAVYDYAASGFDSYIVGASSRLKAIEAENGSLDKFDKATDIGDGSKDKIDITIEGIEVTINGITYELTKKPV
(SEQ ID NO: 62) S.uberis Prophage (ATCC700407) Ply700 Amidase MTDSIQEMRKLQSIPVRYDMGDRYGNDADRDGRIEMDCSSAVSKALGISMTNNTETLQQALPAIGYGKIHDAVDGTFDMQAYDVIIWAPRDGSSSLGAFGHVLIATSPTTAIHCNYGSDGITENDYNYIWEINGRPREIVFRKGVTQTQATVTSQFERELDVNARLTVSDKPYYEATLSEDYYVEAGPRIDSQDKELIKAGTRVRVYPEEKWNGNG
(SEQ ID NO: 63) S. suis SMP LySMP glycosidase and endopeptidase But. B. anthracis Bcp1 PlyB Muramidase But. S. aureus Phi11 and Phi12 Lysine Phi11 Amidase and endopeptidase MQAKLTKNEFIEWLKTSEGKQFNVDLWYGFQCFDYANAGWKVLFGLLLKGLGAKDIPFANNFDGLATVYQNTPDFLAQPGDMVVFGSNYGAGYGHVAWVIEATLDYIIVYEQNWLGGGWTDGIEQPGWGWEKVTRRQHAYDFPMWFIRPNFKSETAPRSVQSPTQAPKKETAKPQPKAVELKIIKDVVKGYDLPKRGSNPKGIVIHNDAGSKGATAEAYRNGLVNAPLSRLEAGIAHSYVSGNTVWQALDESQVGWHTANQIGNKYYYGIEVCQSMGADNATFLKNEQATFQECARLLKKWGLPANRNTIRLHNEFTSTSCPHRSSVLHTGFDPVTRGLLPEDKRLQLKDYFIKQIRAYMDGKIPVATVSNESSASSNTVKPVASAWKRNKYGTYYMEESARFTNGNQPITVRKVGPFLSCPVGYQFQPGGYCDYTEVMLQDGHVWVGYTWEGQRYYLPIRTWNGSAPPNQILGDLWGEIS
(SEQ ID NO: 64) S. aureus ΦH5 LysH5 Amidase and endopeptidase MQAKLTKKEFIEWLKTSEGKQYNADGWYGFQCFDYANAGWKALFGLLLKGVGAKDIPFANNFDGLATVYQNTPDFLAQPGDMVVFGSNYGAGYGHVAWVIEATLDYIIVYEQNWLGGGWTDGVQQPGSGWEKVTRRQHAYDFPMWFIRPNFKSETAPRSVQSPTQASKKETAKPQPKAVELKIIKDVVKGYDLPKRGSNPNFIVIHNDAGSKGATAEAYRNGLVNAPLSRLEAGIAHSYVSGNTVWQALDESQVGWHTANQIGNKYGYGIEVCQSMGADNATFLKNEQATFQECARLLKKWGLPANRNTIRLHNEFTSTSCPHRSSVLHTGFDPVTRGLLPEDKRLQLKDYFIKQIRAYMDGKIPVATVSNDSSASSNTVKPVASAWKRNKYGTYYMEESARFTNGNQPITVRKVGPFLSCPVGYQFQPGGYCDYTEVMLQDGHVWVGYTWEGQRYYLPIRTWNGSAPPNQILGDLWGEIS
(SEQ ID NO: 65) S. warneri ΦWMY LysWMY Amidase and endopeptidase MKTKAQAKSWINSKIGKGIDWDGMYGYQCMDEAVDYIHHVTDGKVTMWGNAIDAPKNNFQGLCTVYTNTPEFRPAYGDVIVWSYGTFATYGHIAIVVNPDPYGDLQYITVLEQNWNGNGIYKTEFATIRTHDYTGVSHFIRPKFADEVKETAKTVNKLSVQKKIVTPKNSVERIKNYVKTSGYINGEHYELYNRGHKPKGVVIHNTAGTASATQEGQRLTNMTFQQLANGVAHVYIDKNTIYETLPEDRIAWHVAQQYGNTEFYGIEVCGSRNTDKEQFLANEQVAFQEAARRLKSWGMRANRNTVRLHHTFSSTECPDMSMLLHTGYSMKNGKPTQDITNKCADYFMKQINAYIDGKQPTSTVVGSSSSNKLKAKNKDKSTGWNTNEYGTLWKKEHATFTCGVRQGIVTRTTGPFTSCPQAGVLYYGQSVNYDTVCKQDGYVWISWTTSDGYDVWMPIRTWDRSTDKVSEIWGTIS
(SEQ ID NO: 66) Streptococci (GBS) ΦNCTC 11261 PlayGBS Muramidase and endopeptidase MATYQEYKSRSNGNAYDIDGSFGAQCWDGYADYCKYLGLPYANCTNTGYARDIWEQRHENGILNYFDEVEVMQAGDVAIFMVVDGVTPYSHVAIFDSDAGGGYGWFLGQNQGGANGAYNIVKIPYSATYPTAFRPKVFKNAVTVTGNIGLNKGDYFIDVSAYQQADLTTTCQQAGTTKTIIKVSESIAWLSDRHQQQANTSDPIGYYHFGRFGGDSALAQREADLFLSNLPSKKVSYLVIDYEDSASADKQANTNAVIAFMDKIASAGYKPIYYSYKPFTLNNIDYQKIIAKYPNSIWIAGYPDYEVRTEPLWEFFPSMDGVRWWQFTSVGVAGGLDKNIVLLADDSSKMDIPKVDKPQELTFYQKLATNTKLDNSNVPYYEATLSTDYYVESKPNASSADKEFIKAGTRVRVYEKVNGWSRINHPESAQWVEDSYLVNATDM
(SEQ ID NO: 67) C. perfringens Φ3626 Ply3626 Amidase But. C. difficile ΦCD27 Lysine CD27 Amidase But. E. faecalis Φ1 PlyV12 Amidase But. A. naeslundii ΦAv-1- Lysine Av-1 Putative amidase/muramidase But. L. gasseri ΦgaY LysgaY Muramidase But. S. aureus ΦSA4 LysSA4 Amidase and endopeptidase But. S. haemolyticus ΦSH2 SH2 Amidase and endopeptidase But. B. thuringiensis ΦBtCS33 PlyBt33 Amidase But. L. monocytogenes ΦP40 PlyP40 Amidase But. L. monocytogenes ΦFWLLm3 LysZ5 Amidase MVKYTVENKIIAGLPKGKLKGANFVIAHETANSKSTIDNEVSYMTRNWQNAFVTHFVGGGGRVVQVANVNYVSWGAGQYANSYSYAQVELCRTSNATTFKKDYEVYCQLLVDLAKKAGIPITLDSGSKTSDKGIKSHKWVADKLGGTTHQDPYAYLSSWGISKAQFASDLAKVSGGGNTGTAPAKPSTPSTNLDKLGLVDYMNAKKMDSSYSNRAKLAKQYGIANYSGTASQNTTLLSKIKGGAPKPSTPAPKPSTSTAKKIYFPPNKGNWSVYPTNKAPVKANAIGAINPTKFGGLTYTIQKDRGNGVYEIQTDQFGRVQVYGAPSTGAVIKK
(SEQ ID NO: 68) B. cereus ΦBPS13 LysBPS13 Amidase MAKREKYIFDVEAEVGKAAKSIKSLEAELSKLQKLNKEIDATGGDRTEKEMLATLKAAKEVNAEYQKMQRILKDLSKYSGKVSRKEFNDSKVINNAKTSVQGGKVTDSFGQMLKNMERQINSVNKQFDNHRKAMVDRGQQYTPHLKTNRKDSQGNSNPSMMGRNKSTTQDMEKAVDKFLNGQNEATTGLNQALYQLKEISKLNRRSESLSRRASASGYMSFQQYSNFTGDRRTVQQTYGGLKTANRERVLELSGQATGISKELDRLNSKKGLTAREGEERKKLMRQLEGIDAELTARKKLNSSLDETTSNMEKFNQSLVDAQVSVKPERGTMRGMMYERAPAIALAIGGAITATIGKLYSEGGNHSKAMRPDEMYVGQQTGAVGANWRPNRTATMRSGLGNHLGFTGQEMMEFQSNYLSANGYHGAEDMKAATTGQATFARATGLGSDEVKDFFNTAYRSGGIDGNQTKQFQNAFLGAMKQSGAVGREKDQLKALNGILSSMSQNRTVSNQDMMRTVGLQSAISSSGVASLQGTKGGALMEQLDNGIREGFNDPQMRVLFGQGTKYQGMGGRAALRKQMEKGISDPDNLNTLIDASKASAGQDPAEQAEVLATLASKMGVNMSSDQARGLIDLQPSGKLTKENIDKVMKEGLKEGSIESAKRDKAYSESKASIDNSSEAATAKQATELNDMGSKLRQANAALGGLPAPLYTAIAAVVAFTAAVAGSALMFKGASWLKGGMASKYGGKGGKGGKGGGTGGGGGAGGAAATGAGAAAGAGGVGAAAAGEVGAGVAAGGAAAGAAAGGSKLAGVGKGFMKGAGKLMLPLGILMGASEIMQAPEEAKGSAIGSAVGGIGGGIAGGAATGAIAGSFLGPIGTAVGGIAGGIAGGFAGSSLGETIGGWFDSGPKEDASAADKAKADASAAALAAAAGTSGAVGSSALQSQMAQGITGAPNMSQVGSMASALGISSGAMASALGISSGQENQIQTMTDKENTNTKKA NEAKKGDNLSYERENISMYERVLTRAEQILAQARAQNGIMGVGGGGTAGAGGGINGFTGGGKLQFLAAGQKWSSSNLQQHDLGFTDQNLTAEDLDKWIDSKAPQGSMMRGMGATFLKAGQEYGLDPRYLIAHAAEESGWGTSKIARDKGNFFGIGAFDDSPYSSAYEFKDGTGSAAERGIMGGAKWISEKYYGKGNTTLDKMKAAGYATNASWAPNIASIMAGAPTGSGSGNVTATINVNVKGDEKVSDKLKNSSDMKKAGKDIGSLLGFYSREMTIA
(SEQ ID NO: 69) S. aureus ΦGH15 LysGH15 Amidase and endopeptidase MAKTQAEINKRLDAYAKGTVDSPYRIKKATSYDPSFGVMEAGAIDADGYYHAQCQDLITDYVLWLTDNKVRTWGNAKDQIKQSYGTGFKIHENKPSTVPKKGWIAVFTSGSYQQWGHIGIVYDGGNTSTFTILEQNWNGYANKKPTKRVDNYYGLTHFIEIPVKAGTTVKKETAKKSASKTPAPKKKATLKVSKNHINYTMDKRGKKPEGMVIHNDAGRSSGQQYENSLANAGYARYANGIAHYYGSEGYVWEAIDAKNQIAWHTGDGTGANSGNFRFAGIEVCQSMSASDAQFLKNEQAVFQFTAEKFKEWGLTPNRKTVRLHMEFVPTACPHRSMVLHTGFNPVTQGRPSQAIMNKLKDYFIKQIKNYMDKGTSSSTVVKDGKTSSASTPATRPVTGSWKKNQYGTWYKPENATFVNGNQPIVTRIGSPFLNAPVGGNLPAGATIVYDEVCIQAGHIWIGYNAYNGDRVYCPVRTCQGVPPNHIPGVAWGVFK
(SEQ ID NO: 70) S. aureus ΦvB SauS-PLA88 HydH5 Endopeptidase and glycosidase But. E. faecalis F168/08 Lys168 Endopeptidase But. E. faecalis ΦF170/08 Lys170 Amidase But. S. aureus ΦP-27/HP P-27/HP indefinite But. C. perfringens ΦSM101 psm Muramidase But. C. sporogenes Φ8074-B1 CS74L Amidase MKIGIDMGHTLSGADYGVVGLRPESVLTREVGTKVIYKLQKLGHVVVNCTVDKASSVSESLYTRYYRANQANVDLFISIHFNATPGGTGTEVYTYAGRQLGEATRIRQEFKSLGLRDRGTKDGSGLAVIRNTKAKAMLVECCFCDNPNDMKLYNSESFSNAIVKGITGKLPNGESGNNNQGGNKVKAVVIYNEGADRRGAEYLADYLNCPTISNSRTFDYSCVEHVYAVGGKKEQYTKYLKTLLSGANRYDTMQQILNFINGGK
(SEQ ID NO: 71) S. typhimurium ΦSPN1S SPN1S glycosidase MDINQFRRASGINEQLAARWFPHITTAMNEFGITKPDDQAMFIAQVGHESGGFTRLQENFNYSVNGLSGFIRAGRITPDQANALGRKTYEKSLPLERQRAIANLVYSKRMGNNGPGDGWNYRGRGLIQITGLNNYRDCGNGLKVDLVAQPELLAQDEYAARSAAWFFSSKGCMKYTGDLVRVTQIINGGQNGIDDRRTRYAAARKVLAL
(SEQ ID NO: 72) C. michiganensis ΦCMP1 CMP1 Peptidase But. C. michiganensis ΦCN77 CN77 Peptidase MGYWGYPNGQIPNDKMALYRGCLLRADAAAQAYALQDAYTRATGKPLVILEGYRDLTRQKYLRNLYLSGRGNIAAVPGLSNHGWGLACDFAAPLNSSGSEEHRWMRQNAPLFGFDWARGKADNEPWHWEYGNVPVSRWASLDVTPIDRNDMADITEGQMQRIAVILLDTEIQTPLGPRLVKHALGDALLLGQANANSIAEVPDKTWDVLVDHPLAKNEDGTPLKVRLGDVAKYEPLEHQNTRDAIAKLGTLQFTDKQLATIGAGVKPIDEASLVKKIVDGVRALFGRAAA
(SEQ ID NO: 73) A. baumannii ΦAB2 LysAB2 glycosidase MILTKDGFSIIRNELFGGKLDQTQVDAINFIVAKATESGLTYPEAAYLLATIYHETGLPSGYRTMQPIKEAGSDSYLRSKKYYPYIGYGYVQLTWKENYERIGKLIGVDLIKNPEKALEPLIAIQIAIKGMLNGWFTGVGFRRKRPVSKYNKQQYVAARNIINGKDKAELIAKYAIIFERALRSL
(SEQ ID NO: 74) B. cereus ΦB4 LysB4 Endopeptidase MAMALQTLIDKANRKLNVSGMRKDVADRTRAVITQMHAQGIYICVAQGFRSFAEQNALYAQGRTKPGSIVTNARGGQSNHNYGVAVDLCLYTQDGSDVIWTVEGNFRKVIAAMKAQGFKWGGDWVSFKDYPHFELYDVVGGQKPPADNGGAVDNGGGSGSTGGSGGGSTGGGSTGGGYDSSWFTKETGTFVTNTSIKLRTAPFTSADVIATLPAGSPVNYNGFGIEYDGYVWIRQPRSNGYGYLATGESKGGKRQNYWGTFK
(SEQ ID NO: 75) P. aeruginosa ΦKMV KMV45 indefinite But. C. tyrobutyricum ΦCTP1 Ctp1l glycosidase MKKIADISNLNGNVDVKLLFNLGYIGIIAKASEGGTFVDKYYKQNYTNTKAQGKITGAYHFANFSTIAKAQQEANFFLNCIAGTTPDFVVLDLEQQCTGDITDACLAFLNIVAKKFKCVVYCNSSFIKEHLNSKICAYPLWIANYGVATPAFTLWTKYAMWQFTEKGQVSGISGYIDFSYITDEFIKYIKGEDEVENLVVYNDGADQRAAEYLADRLACPTINNARKFDYSNVKNVYAVGGNKEQYTSYLTTLIAGSTRYTTMQAVLDYIKNLK
(SEQ ID NO: 76) P. aeruginosa ΦEL EL188 Transglycosylase But. P. aeruginosa ΦKZ KZ144 Transglycosylase But. S. aureus Ply187 Cell wall hydrolase MALPKTGKPTAKQVVDWAINLIGSGVDVDGYYGRQCWDLPNYIFNRYWNFKTPGNARDMAWYRYPEGFKVFRNTSDFVPKPGDIAVWTGGNYNWNTWGHTGIVVGPSTKSYFYSVDQNWNNSNSYVGSPAAKIKHSYFGVTHFVRPAYKAEPKPTPPAQNNPAPKDPEPSKKPESNKPIYKVVTKILFTTAHIEHVKANRFVHYITKSDNHNNKPNKIVIKNTNTALSTIDVYRYRDELDKDEIPHFFVDRLNVWACRPIEDSINGYHDSVVLSITETRTALSDNFKMNEIECLSLAESILKANNKKMSASNIIVDNKAWRTFKLHTGKDSLKSSSFTSKDYQKAVNELIKLFNDKDKLLNNKPKDVVERIRIRTIVKENTKFVPSELKPRNNIRDKQDSKIDRVINNYTLKQALNIQYKLNPKPQTSNGVSWYNASVNQIKSAMDTTKIFNNNVQVYQFLKLNQYQGIPVDKLNKLLVGKGTLANQGHAFADGCKKYNINEIYLIAHRFLESANGTSFFASGKTGVYNYFGIGAFDNNPNNAMAFARSHGWTSPTKAIIGGAEFVGKGYFNVGQNTLYRMRWNPQKPGTHQYATDISWAKVQAQMISAMYKEIGLTGDYFIYDQYKK
(SEQ ID NO: 77) P. fluorescens ΦOBP OBPgp279 glycosidase But. L. monocytogenes ΦP35 PlyP35 Amidase MARKFTKAELVAKAEKKVGGLKPDVKKAVLSAVKEAYDRYGIGIIVSQGYRSIAEQNGLYAQGRTKPGNIVTNAKGGQSNHNFGVAVDFAIDLIDDGKIDSWQPSATIVNMMKRRGFKWGGDWKSFTDLPHFEACDWYRGERKYKVDTSEWKKKENINIVIKDVGYFQDKPQFLNSKSVRQWKHGTKVKLTKHNSHWYTGVVKDGNKSVRGYIYHSMAKVTSKNSDGSVNATINAHAFCWDNKKLNGGDFINLKRGFKGITHPASDGFYPLYFASRKKTFYIPRYMFDIKK
(SEQ ID NO: 78) L. fermentum ΦPYB5 Lyb5 Muramidase But. S. pneumoniae ΦCP-7 cpl-7 Muramidase MVKKNDLFVDVASHQGYDISGILEEAGTTNTIIKVSESTSYLNPCLSAQVSQSNPIGFYHFAWFGGNEEEAEAEARYFLDNVPTQVKYLVLDYEDHASASVQRNTTACLRFMQIIAEAGYTPIYYSYKPFTLDNVDYQQILAQFPNSLWIAGYGLNDGTANFEYFPSMDGIRWWQYSSNPFDKNIVLLDDEKEDNINNENTLKSLTTVANEVIQGLWGNGQERYDSLANAGYDPQAVQDKVNEILNAREIADLTTVANEVIQGLWGNGQERYDSLANAGYDPQAVQDKVNEILNAREIADLTTVANEVIQGLWGNGQERYDSLANAGYDPQAVQDKVNELLS
(SEQ ID NO: 79) P. chlororaphis201 Φ2-1 2012-1gp229 glycosidase But. S. enterica ΦPVP-SE1) PVP-SE1gp146 glycosidase But. Corynebacterium ΦBFK20 BKF20 Amidase But. E. faecalis ΦEFAP-1 EFAL-1 Amidase MKLKGILLSVVTTFGLLFGATNVQAYEVNNEFNLQPWEGSQQLAYPNKIILHETANPRATGRNEATYMKNNWFNAHTTAIVGDGGIVYKVAPEGNVSWGAGNANPYAPVQIELQHTNDPELFKANYKAYVDYTRDMGKKFGIPMTLDQGGSLWEKGVVSHQWVTDFVWGDHTDPYGYLAKMGISKAQLAHDLANGVSGNTATPTPKPDKPKPTQPSKPSNKKRFNYRVDGLEYVNGMWQIYNEHLGKIDFNWTENGIPVEVVDKVNPATGQPTKDQVLKVGDYFNFQENSTGVVQEQTPYMGYTLSHVQLPNEFIWLFTDSKQALMYQ
(SEQ ID NO: 80) Lactobacilli LamdaSA2 LysA, LysA2 and LysgaY indefinite But. S. aureus SAL-1 indefinite But.

In some instances, lysine is a functionally active variant of the lysines described herein. In some cases, the lysine variant is at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83 %, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical , for example, in a certain area or in the entire sequence, the sequence of the lysine described in this document, or naturally occurring lysine.

In some cases, lysine can be created using bioengineering to modulate its biological activity, for example, increase, or decrease, or regulation, or to determine its target microorganism. In some cases, lysine is produced by the translation apparatus (eg, ribosome, etc.) of the microbial cell. In some cases, lysine is synthesized chemically. In some cases, lysine is formed from a polypeptide precursor. The precursor polypeptide may be cleaved (eg, processed by a protease) to form the lysine polypeptide itself. Thus, in some cases, lysine is produced from a precursor polypeptide. In some instances, lysine includes a polypeptide that has undergone post-translational modifications, such as cleavage or the addition of one or more functional groups.

The lysines described herein may be formulated for any of the uses described herein. The compositions disclosed herein may contain any number or type (e.g., classes) of lysines, such as at least about any of 1 lysine, 2, 3, 4, 5, 10, 15, 20 or more lysines . The appropriate concentration of each lysine in the composition depends on factors such as potency, stability of the lysine, the amount of different lysines, composition and methods of application of the composition. In some cases, the concentration of each lysine in the liquid composition is from about 0.1 ng/ml to about 100 mg/ml. In some instances, the concentration of each lysine in the solid composition is from about 0.1 ng/g to about 100 mg/g. In some cases, if the composition contains at least two types of lysines, then the concentration of each type of lysins may be the same or different.

A modulator comprising a lysine described herein may be contacted with a target host in an amount and for a time sufficient to: host-target; (b) achieving a target level (eg, a predetermined or threshold level) of lysine concentration within the gut of the target host; (c) reaching a target level (eg, a predetermined or threshold level) of lysine concentration within the target host bacteriocyte; (d) modulating the level or activity of one or more microorganisms (eg, endosymbionts) in the target host or/and (e) modulating the fitness of the target host.

Antimicrobial peptides

The modulator described herein may include an antimicrobial peptide (AMP). Any AMP suitable for inhibiting a microorganism residing in the host may be used. AMPs are a diverse group of molecules that are divided into subgroups based on their amino acid composition and structure. AMPs can be obtained from or produced in any organism that naturally produces AMP, including AMPs derived from plants (e.g., copsin), insects (e.g., drozocin, scorpion peptide (e.g., Uy192, UyCT3, D3, D10, Uy17, Uy192), mastoparan, poneratoxin, cecropin, moricin, mellitin), frogs (eg magainin, dermaseptin, aurein) and mammals (eg cathelicidins, defensins and protegrins). For example, AMP may be a scorpion peptide such as Uy192 (5'-FLSTIWNGIKGLL-3'; SEQ ID NO: 221), UyCT3 (5'-LSAIWSGIKSLF-3; SEQ ID NO: 222), D3 (5'-LWGKLWEGVKSLI -3'; SEQ ID NO: 223) and D10 (5'-FPFLKLSLKIPKSAIKSAIKRL-3'; SEQ ID NO: 224), Uy17 (5'-ILSAIWSGIKGLL-3'; SEQ ID NO: 225) or a combination thereof. Other non-limiting examples of AMP are shown in Table 6.

Table 6. Examples of antimicrobial peptides

Type of Characteristic Illustrative AMP Subsequence Anionic peptides Rich in glutamic and aspartic acid Dermsidin SSLLEKGLDGAKKAVGGLGKLGKDAVEDLESVGKGAVHDVKDVLDSVL
(SEQ ID NO: 81) Linear cationic α-helical peptides Do not have cysteine Cecropin A KWKLFKKIEKVGQNIRDGIIKAGPAVAVVGQATQIAK
(SEQ ID NO: 82) Andropin MKYFSVLVVLTLILAIVDQSDAFINLLDKVEDALHTGAQAGFKLIRPVERGATPKKSEKPEK
(SEQ ID NO: 83) Moricin MNILKFFFVFIVAMSLVSCSTAAPAKIPIKKAIKTVGKAVGKGLRAINIASTANDVFNFLKPKKRKH
(SEQ ID NO: 84) Ceratotoxin MANLKAVFLICIVAFIALQCVVAEPAAEDSVVVKRSIGSALKKALPVAKKIGKIALPIAKAALPVAAGLVG
(SEQ ID NO: 85) Cationic peptides enriched with specific amino acids Rich in proline, arginine, phenylalanine, glycine, tryptophan Abetsin MKVVIFIFALLATICAAFAYVPLPNVPQPGRRPFPTFPGQGPFNPKIKWPQGY
(SEQ ID NO: 86) Apidecins KNFALAILVVTFVVAVFGNTNLDPPTRPTRLRREAKPEAEPGNNRPVYIPQPRPPHPRLRREAEPEAEPGNNRPVYIPQPRPPHPRLRREAELEAEPGNNRPVYISQPRPPHPRLRREAEPEAEPGNNRPVYIPQPRPPHPRLRREAELEAEPGNNRPVYISQPRPPHPRLRREAEPEAEPGNNRPVYIPQPRPPHPRLRREAEPEAEPGNNRPVYIPQPRPPHPRLRREAEPEAEPGNNRPVYIPQPRPPHPRLRREAKPEAKPGNNRPVYIPQPRPPHPRI
(SEQ ID NO: 87) Profinin METQRASLCLGRWSLWLLLALVVPSASAQALSYREAVLRAVDRLNEQSSEANLYRLLELDQPPKADEDPGTPKPVSFTVKETVCPRPTRRPPELCDFKENGRVKQCVGTVTLDQIKDPLDITCNEGVRRFPWWWPPNVPGPRFPPPNVPGPRFPPPNFPGPRFPPPNFPGPRFPPPNFPGPPFPPPIFPGPWFPPPPPFRPPPFGPPRFPGRR
(SEQ ID NO: 88) Indolicidin MQTQRASLSLGRWSLWLLLGLVVPSASAQALSYREAVLRAVDQLNELSSEANLYRLLELDPPPKDNEDLGTRKPVSFTVKETVCPRTIQQPAEQCDFKEKGRVKQCVGTVTLDPSNDQFDLNCNELQSVILPWKWPWWPWRRG
(SEQ ID NO: 89) Anionic and cationic peptides that contain cysteine and form disulfide bonds Contain 1-3 disulfide bonds Protegrin METQRASLCLGRWSLWLLLALVVPSASAQALSYREAVLRAVDRLNEQSSEANLYRLLELDQPPKADEDPGTPKPVSFTVKETVCPRPTRQPPELCDFKENGRVKQCVGTVTLDQIKDPLDITCNEVQGVRGGRLCYCRRRFCVCVGRG
(SEQ ID NO: 90) Tachyplesins KWCFRVCYRGYRRCR
(SEQ ID NO: 91) Defensins MKCATIVCTIAVVLAATLLNGSVQAAPQEEAALSGGANLNTLLDELPEETHHAALENYRAKRATCDLASGFGVGSSLCAAHCIARRYRGGYCNSKAVCVCRN
(SEQ ID NO: 92) Drosomycin MMQIKYLFALFAVLMLVVLGANEADADCLSGRYKGPCAVWDNETCRRVCKEEGRSSGHCSPSLKCWCEGC
(SEQ ID NO: 93)

AMP can be active against any number of target microorganisms. In some cases, AMP may be characterized by forms of antibacterial and/or antifungal activity. In some cases, AMP may have narrow spectrum biological activity or broad spectrum biological activity. For example, some AMPs target and kill only a few types of bacteria or fungi, while others are active against both gram-negative and gram-positive bacteria, as well as fungi.

In addition, AMP can function through a number of known mechanisms of action. For example, the cytoplasmic membrane is a common target for AMP, but AMP can also interfere with DNA and protein synthesis, protein folding, and cell wall synthesis. In some cases, AMPs with a total cationic charge and amphipathic nature destroy bacterial membranes, leading to cell lysis. In some cases, AMP can enter cells and interact with an intracellular target to disrupt DNA, RNA, protein, or cell wall synthesis. In addition to killing microorganisms, AMPs have demonstrated a number of immunomodulatory functions that are involved in clearing infection, including the ability to alter host gene expression, act as chemokines and/or induce chemokine production, inhibit lipopolysaccharide-induced pro-inflammatory cytokine production, promote wound healing, and modulate responses. dendritic cells and cells involved in the adaptive immune response.

In some cases, AMP is a functionally active variant of the AMPs described in this document. In some cases, the AMP variant is at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83 %, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical , for example, in a certain area or in the entire sequence, the sequence of the AMP described in this document, or AMP of natural origin.

In some cases, an AMP can be bioengineered to modulate its biological activity, such as increasing or decreasing or regulating it, or to determine its target microorganism. In some cases, AMP is produced by the translation apparatus (eg, ribosome, etc.) of the cell. In some cases, AMP is synthesized chemically. In some cases, AMP is formed from a polypeptide precursor. The precursor polypeptide may be cleaved (eg, processed by a protease) to form the AMP polypeptide itself. Thus, in some cases, AMP is produced from a precursor polypeptide. In some cases, AMP includes a polypeptide that has undergone post-translational modifications, such as cleavage or the addition of one or more functional groups.

The AMPs described herein may be formulated for any of the uses described herein. The compositions disclosed herein may contain any number or type (eg, classes) of AMP, such as at least about any of 1 AMP, 2, 3, 4, 5, 10, 15, 20 or more AMP . For example, the compositions may contain an AMP cocktail (eg, a scorpion peptide cocktail, eg UyCT3, D3, D10 and Uy17). The appropriate concentration of each AMP in the composition depends on such factors as the potency, stability of the AMP, the amount of different AMPs in the composition, the composition and methods of application of the composition. In some cases, the concentration of each AMP in the liquid composition is from about 0.1 ng/ml to about 100 mg/ml. In some instances, the concentration of each AMP in the solid composition is from about 0.1 ng/g to about 100 mg/g. In some cases, if the composition contains at least two types of AMP, then the concentration of each type of AMP may be the same or different.

A modulator comprising an AMP described herein may be contacted with a target host in an amount and for a time sufficient to: host-target; (b) achieving a target level (eg, a predetermined or threshold level) of AMP concentration within the gut of the target host; (c) achieving a target level (eg, a predetermined or threshold level) of AMP concentration within the target host bacteriocyte; (d) modulating the level or activity of one or more microorganisms (eg, endosymbionts) in the target host or/and (e) modulating the fitness of the target host.

C-rich nodule peptides

The modulator described herein may include a C-rich nodule peptide (NCR peptide). NCR peptides are produced in certain species of leguminous plants and play an important role in the mutualistic nitrogen-fixing symbiosis of plants with bacteria from the Rhizobiaceae family (nodule bacteria), which leads to the formation of root nodules in which plant cells contain thousands of intracellular endosymbionts. NCR peptides have antimicrobial properties by directing the irreversible process of terminal bacterial differentiation, such as permeabilization of the bacterial membrane, disruption of cell division, or inhibition of protein synthesis. For example, Medicago truncatula nodule cells infected with Sinorhizobium meliloti produce hundreds of NCR peptides that control the irreversible process of bacterial differentiation into large polyploid nitrogen-fixing bacteroids. Non-limiting examples of NCR peptides are shown in Table 7.

Table 7 Examples of NCR peptides

Name Peptide sequence Producer >gi|152218086|gb|ABS31477.1| NCR 340 MTKIVVFIYVVILLLTIFHVSAKKKRYIECETHEDCSQVFMPPFVMRCVIHECKIFNGEHLRY
(SEQ ID NO: 94) Medicago truncatula >gi|152218084|gb|ABS31476.1| NCR 339 MAKIMKFVYNMIPFLSIFIITLQVNVVVCEIDADCPQICMPPYEVRCVNHRCGWVNTDDSLFLTQEFTRSKQYIIS
(SEQ ID NO: 95) Medicago truncatula >gi|152218082|gb|ABS31475.1| NCR 338 MYKVVESIFIRYMHRKPNMTKFFKFVYTMFILISLFLVVTNANAHNCTDISDCSSNHCSYEGVSLCMNGQCICIYE
(SEQ ID NO: 96) Medicago truncatula >gi|152218080|gb|ABS31474.1| NCR 337 MVETLRLFYIMILFVSLCLVVVDGESKLEQTCSEDFECYIKNPHVPFGHLRCFEGFCQQLNGPA
(SEQ ID NO: 97) Medicago truncatula >gi|152218078|gb|ABS31473.1| NCR 336 MAKIVNFVYSMIVFLFLFLVATKAARGYLCVTDSHCPPHMCPPGMEPRCVRRMCKCLPIGWRKYFVP
(SEQ ID NO: 98) Medicago truncatula >gi|152218076|gb|ABS31472.1| NCR 335 MQIGKNMVETPKLDYVIIFFFLYFFFRQMIILRLNTTFRPLNFKMLRFWGQNRNIMKHRGQKVHFSLILSDCKTNKDCPKLRRANVRCRKSYCVPI
(SEQ ID NO: 99) Medicago truncatula >gi|152218074|gb|ABS31471.1| NCR 334 MLRLYLVSYFLLKRTLLVSYFSYFSTYIIECKTDDNDCPISQLKIYAWKCVKNGCHLFDVIPMMYE
(SEQ ID NO: 100) Medicago truncatula >gi|152218072|gb|ABS31470.1| NCR 333 MAEILKFVYIVILFVSLLLIVVASERECVTDDDCEKLYPTNEYRMMCDSGYCMNLLNGKIIYLLCLKKKKFLIIISVLL
(SEQ ID NO: 101) Medicago truncatula >gi|152218070|gb|ABS31469.1| NCR 332 MAEIIKFVYIMILCVSLLLIEVAGEECVTDADCDKLYPDIRKPLMCSIGECYSLYKGKFSLSIISKTSFSLMVYNVVTLVICLRLAYISLLLKFL
(SEQ ID NO: 102) Medicago truncatula >gi|152218068|gb|ABS31468.1| NCR 331 MAEILKDFYAMNLFIFLIILPAKIRGETLSLTHPKCHHIMLPSLFITEVFQRVTDDGCPKPVNHLRVVKCIEHICEYGYNYRPDFASQIPESTKMPRKRE
(SEQ ID NO: 103) Medicago truncatula >gi|152218066|gb|ABS31467.1| NCR 330 MVEILKNFYAMNLFIFLIILAVKIRGAHFPCVTDDDCPKPVNKLRVIKCIDHICQYARNLPDFASEISESTKMPCKGE
(SEQ ID NO: 104) Medicago truncatula >gi|152218064|gb|ABS31466.1| NCR 329 MFHAQAENMAKVSNFVCIMILFLALFFITMNDAARFECREDSHCVTRIKCVLPRKPECRNYACGCYDSNKYR
(SEQ ID NO: 105) Medicago truncatula >gi|152218062|gb|ABS31465.1| NCR 328 MQMRQNMATILNFVFVIILFISLLLVVTKGYREPFSSFTEGPTCKEDIDCPSISCVNPQVPKCIMFECHCKYIPTTLK
(SEQ ID NO: 106) Medicago truncatula >gi|152218060|gb|ABS31464.1| NCR 327 MATILMYVYITILFISILTVLTEGLYEPLYNFRRDPDCRRNIDCPSYLCVAPKVPRCIMFECHCKDIPSDH
(SEQ ID NO: 107) Medicago truncatula >gi|152218058|gb|ABS31463.1| NCR 326 MTTSLKFVYVAILFLSLLLVVMGGIRRFECRQDSDCPSYFCEKLTVPKCFWSKCYCK
(SEQ ID NO: 108) Medicago truncatula >gi|152218056|gb|ABS31462.1| NCR 325 MTTSLKFVYVAILFLSLLLVVMGGIRKKECRQDSDCPSYFCEKLTIAKCIHSTCLCK
(SEQ ID NO: 109) Medicago truncatula >gi|152218054|gb|ABS31461.1| NCR 324 MQIGKNMVETPKLVYFIILFLSIFLCITVSNSSFSQIFNSACKTDKDCPKFGRVNVRCRKGNCVPI
(SEQ ID NO: 110) Medicago truncatula >gi|152218046|gb|ABS31457.1| NCR 320 MTAILKKFINAVFLFIVLFLATTNVEDFVGGSNDECVYPDVFQCINNICKCVSHHRT
(SEQ ID NO: 111) Medicago truncatula >gi|152218044|gb|ABS31456.1| NCR 319 MQKRKNMAQIIFYVYALIILFSPFLAARLVFVNPEKPCVTDADCDRYRHESAIYSDMFCKDGYCFIDYHHDPYP
(SEQ ID NO: 112) Medicago truncatula >gi|152218042|gb|ABS31455.1| NCR 318 MQMRKNMAQILFYVYALLILFTPFLVARIMVVNPNNPCVTDADCQRYRHKLATRMICNQGFCLMDFTHDPYAPSLP
(SEQ ID NO: 113) Medicago truncatula >gi|152218040|gb|ABS31454.1| NCR 317 MNHISKFVYALIIFLSIYLVVLDGLPISCKDHFECRRKINILRCIYRQEKPMCINSICTCVKLL
(SEQ ID NO: 114) Medicago truncatula >gi|152218038|gb|ABS31453.1| NCR 316 MQREKNMAKIFEFVYAMIIFILLFLVEKNVVAYLKFECKTDDDCQKSLLKTYVWKCVKNECYFFAKK
(SEQ ID NO: 115) Medicago truncatula >gi|152218036|gb|ABS31452.1| NCR 315 MAGIIKFVHVLIIFLSLFHVVKNDDGSFCFKDSDCPDEMCPSPLKEMCYFLQCKCGVDTIA
(SEQ ID NO: 116) Medicago truncatula >gi|152218034|gb|ABS31451.1| NCR 314 MANTHKLVSMILFIFLFLASNNVEGYVNCETDADCPPSTRVKRFKCVKGECRWTRMSYA
(SEQ ID NO: 117) Medicago truncatula >gi|152218032|gb|ABS31450.1| NCR 313 MQRRKKKAQVVMFVHDLIICIYLFIVITTRKTDIRCRFYYDCPRLEYHFCECIEDFCAYIRLN
(SEQ ID NO: 118) Medicago truncatula >gi|152218030|gb|ABS31449.1| NCR 312 MAKVYMFVYALIIFVSPFLLATFRTRLPCEKDDDCPEAFLPPVMKCVNRFCQYEILE
(SEQ ID NO: 119) Medicago truncatula >gi|152218028|gb|ABS31448.1| NCR 310 MIKQFSVCYIQMRRNMTTILKFPYIMVICLLLLHVAAYEDFEKEIFDCKKDGDCDHMCVTPGIPKCTGYVCFCFENL
(SEQ ID NO: 120) Medicago truncatula >gi|152218026|gb|ABS31447.1| NCR 309 MQRSRNMTTIFKFAYIMIICVFLLNIAAQEIENGIHPCKKNEDCNHMCVMPGLPWCHENNLCFCYENAYGNTR
(SEQ ID NO: 121) Medicago truncatula >gi|152218024|gb|ABS31446.1| NCR 304 MTIIIKFVNVLIIFLSLFHVAKNDDNKLLLSFIEEGFLCFKDSDCPYNMCPSPLKEMCYFIKCVCGVYGPIRERRLYQSHNPMIQ
(SEQ ID NO: 122) Medicago truncatula >gi|152218022|gb|ABS31445.1| NCR 303 MRKNMTKILMIGYALMIFIFLSIAVSITGNLARASRKKPVDVIPCIYDHDCPRKLYFLERCVGRVCKYL
(SEQ ID NO: 123) Medicago truncatula >gi|152218020|gb|ABS31444.1| NCR 301 MAHKLVYAITLFIFLFLIANNIEDDIFCITDNDCPPNTLVQRYRCINGKCNLSFVSYG
(SEQ ID NO: 124) Medicago truncatula >gi|152218018|gb|ABS31443.1| NCR 300 MDETLKFVYILILFVSLCLVVADGVKNINRECTQTSDCYKKYPFIPWGKVRCVKGRCRLDM
(SEQ ID NO: 125) Medicago truncatula >gi|152218016|gb|ABS31442.1| NCR 290 MAKIIKFVYVLAIFFSLFLVAKNVNGWTCVEDSDCPANICQPPMQRMCFYGECACVRSKFCT
(SEQ ID NO: 126) Medicago truncatula >gi|152218014|gb|ABS31441.1| NCR 289 MVKIIKFVYFMTLFLSMLLVTTKEDGSVECIANIDCPQIFMLPFVMRCINFRCQIVNSEDT
(SEQ ID NO: 127) Medicago truncatula >gi|152218012|gb|ABS31440.1| NCR 286 MDEILKFVYTLIIFFSLFFAANNVDANIMNCQSTFDCPRDMCSHIRDVICIFKKCKCAGGRYMPQVP
(SEQ ID NO: 128) Medicago truncatula >gi|152218008|gb|ABS31438.1| NCR 278 MQRRKNMANNHMLIYAMIICLFPYLVVTFKTAITCDCNEDCLNFFTPLDNLKCIDNVCEVFM
(SEQ ID NO: 129) Medicago truncatula >gi|152218006|gb|ABS31437.1| NCR 266 MVNILKFIYVIIFFILMFFVLIDVDGHVLVECIENRDCEKGMCKFPFIVRCLMDQCKCVRIHNLI
(SEQ ID NO: 130) Medicago truncatula >gi|152218004|gb|ABS31436.1| NCR 265 MIIQFSIYYMQRRKLNMVEILKFSHALIIFLFLSALVTNANIFFCSTDEDCTWNLCRQPWVQKCRLHMCSCEKN
(SEQ ID NO: 131) Medicago truncatula >gi|152218002|gb|ABS31435.1| NCR 263 MDEVFKFVYVMIIFPFLILDVATNAEKIRRCFNDAHCPPDMCTLGVIPKCSRFTICIC
(SEQ ID NO: 132) Medicago truncatula >gi|152218000|gb|ABS31434.1| NCR 244 MHRKPNMTKFFKFVYTMFILISLFLVVTNANANNCTDTSDCSSNHCSYEGVSLCMNGQCICIYE
(SEQ ID NO: 133) Medicago truncatula >gi|152217998|gb|ABS31433.1| NCR 239 MQMKKMATILKFVYLIILLIYPLLVVTEESHYMKFSICKDDTDCPTLFCVLPNVPKCIGSKCHCKLMVN
(SEQ ID NO: 134) Medicago truncatula >gi|152217996|gb|ABS31432.1| NCR 237 MVETLRLFYIMILFVSLYLVVVDGVSKLAQSCSEDFECYIKNPHAPFGQLRCFEGYCQRLDKPT
(SEQ ID NO: 135) Medicago truncatula >gi|152217994|gb|ABS31431.1| NCR 228 MTTFLKVAYIMIICVFVLHLAAQVDSQKRLHGCKEDRDCDNICSVHAVTKCIGNMCRCLANVK
(SEQ ID NO: 136) Medicago truncatula >gi|152217992|gb|ABS31430.1| NCR 224 MRINRTPAIFKFVYTIIIYLFLLRVVAKDLPFNICEKDEDCLEFCAHDKVAKCMLNICFCF
(SEQ ID NO: 137) Medicago truncatula >gi|152217990|gb|ABS31429.1| NCR 221 MAEILKILYVFIIFLSLILAVISQHPFTPCETNADCKCRNHKRPDCLWHKCYCY
(SEQ ID NO: 138) Medicago truncatula >gi|152217988|gb|ABS31428.1| NCR 217 MRKSMATILKFVYVIMLFIYSLFVIESFGHRFLIYNNCKNDTECPNDCGPHEQAKCILYACYCVE
(SEQ ID NO: 139) Medicago truncatula >gi|152217986|gb|ABS31427.1| NCR 209 MNTILKFIFVVFLFLSIFLSAGNSKSYGPCTTLQDCETHNWFEVCSCIDFECKCWSLL
(SEQ ID NO: 140) Medicago truncatula >gi|152217984|gb|ABS31426.1| NCR 206 MAEIIKFVYIMILCVSLLLIAEASGKECVTDADCENLYPGNKKPMFCNNTGYCMSLYKEPSRYM
(SEQ ID NO: 141) Medicago truncatula >gi|152217982|gb|ABS31425.1| NCR 201 MAKIIKFVYIMILCVSLLLIVEAGGKECVTDVDCEKIYPGNKKPLICSTGYCYSLYEEPPRYHK
(SEQ ID NO: 142) Medicago truncatula >gi|152217980|gb|ABS31424.1| NCR 200 MAKVTKFGYIIIHFLSLFFLAMNVAGGRECHANSHCVGKITCVLPQKPECWNYACVCYDSNKYR
(SEQ ID NO: 143) Medicago truncatula >gi|152217978|gb|ABS31423.1| NCR 192 MAKIFNYVYALIMFLSLFLMGTSGMKNGCKHTGHCPRKMCGAKTTKCRNNKCQCV
(SEQ ID NO: 144) Medicago truncatula >gi|152217976|gb|ABS31422.1| NCR 189 MTEILKFVCVMIIFISSFIVSKSLNGGGKDKCFRDSDCPKHMCPSSLVAKCINRLCRCRRPELQVQLNP
(SEQ ID NO: 145) Medicago truncatula >gi|152217974|gb|ABS31421.1| NCR 187 MAHIIMFVYALIYALIIFSSLFVRDGIPCLSDDECPEMSHYSFKCNNKICEYDLGEMSDDDYYLEMSRE
(SEQ ID NO: 146) Medicago truncatula >gi|152217972|gb|ABS31420.1| NCR 181 MYREKNMAKTLKFVYVIVLFLSLFLAAKNIDGRVSYNSFIALPVCQTAADCPEGTRGRTYKCINNKCRYPKLLKPIQ
(SEQ ID NO: 147) Medicago truncatula >gi|152217970|gb|ABS31419.1| NCR 176 MAHIFNYVYALLVFLSLFLMVTNGIHIGCDKDRDCPKQMCHLNQTPKCLKNICKCV
(SEQ ID NO: 148) Medicago truncatula >gi|152217968|gb|ABS31418.1| NCR 175 MAEILKCFYTMNLFIFLIILPAKIREHIQCVIDDDCPKSLNKLLIIKCINHVCQYVGNLPDFASQIPKSTKMPYKGE
(SEQ ID NO: 149) Medicago truncatula >gi|152217966|gb|ABS31417.1| NCR 173 MAYISRIFYVLIIFLSLFFVVINGVKSLLLIKVRSFIPCQRSDDCPRNLCVDQIIPTCVWAKCKCKNYND
(SEQ ID NO: 150) Medicago truncatula >gi|152217964|gb|ABS31416.1| NCR 172 MANVTKFVYIAIYFLSLFFIAKNDATATFCHDDSHCVTKIKCVLPRTPQCRNEACGCYHSNKFR
(SEQ ID NO: 151) Medicago truncatula >gi|152217962|gb|ABS31415.1| NCR 171 MGEIMKFVYVMIIYLFMFNVATGSEFIFTKKLTSCDSSKDCRSFLCYSPKFPVCKRGICECI
(SEQ ID NO: 152) Medicago truncatula >gi|152217960|gb|ABS31414.1| NCR 169 MGEMFKFIYTFILFVHLFLVVIFEDIGHIKYCGIVDDCYKSKKPLFKIWKCVENVCVLWYK
(SEQ ID NO: 153) Medicago truncatula >gi|152217958|gb|ABS31413.1| NCR 165 MARTLKFVYSMILFLSLFLVANGLKIFCIDVADCPKDLYPLLYKCIYNKCIVFTRIPPPFDWI
(SEQ ID NO: 154) Medicago truncatula >gi|152217956|gb|ABS31412.1| NCR 159 MANITKFVYIAILFLSLFFIGMNDAAILECREDSHCVTKIKCVLPRKPECRNNACTCYKGGFSFHH
(SEQ ID NO: 155) Medicago truncatula >gi|152217954|gb|ABS31411.1| NCR 147 MQRVKKMSETLKFVYVLILFISIFHVVIVCDSIYFPVSRPCITDKDCPNMKHYKAKCRKGFCISSRVR
(SEQ ID NO: 156) Medicago truncatula >gi|152217952|gb|ABS31410.1| NCR 146 MQIRKIMSGVLKFVYAIILFLFLFLVAREVGGLETIECETDGDCPRSMIKMWNKNYRHKCIDGKCEWIKKLP
(SEQ ID NO: 157) Medicago truncatula >gi|152217950|gb|ABS31409.1| NCR 145 MFVYDLILFISLILVVTGINAEADTSCHSFDDCPWVAHHYRECIEGLCAYRILY
(SEQ ID NO: 158) Medicago truncatula >gi|152217948|gb|ABS31408.1| NCR 144 MQRRKKSMAKMLKFFFAIILLLSLFLVATEVGGAYIECEVDDDCPKPMKNSHPDTYYKCVKHRCQWAWK
(SEQ ID NO: 159) Medicago truncatula >gi|152217946|gb|ABS31407.1| NCR 140 MFVYTLIIFLFPSHVITNKIAIYCVSDDDCLKTFTPLDLKCVDNVCEFNLRCKGKCGERDEKFVFLKALKKMDQKLVLEEQGNAREVKIPKKLLFDRIQVPTPATKDQVEEDDYDDDDEEEEEEEDDVDMWFHLPDVVCH
(SEQ ID NO: 160) Medicago truncatula >gi|152217944|gb|ABS31406.1| NCR 138 MAKFSMFVYALINFLSLFLVETAITNIRCVSDDDDCPKVIKPLVMKCIGNYCYFFMIYEGP
(SEQ ID NO: 161) Medicago truncatula >gi|152217942|gb|ABS31405.1| NCR 136 MAHKFVYAIILFIFLFLVAKNVKGYVVCRTVDDCPPDTRDLRYRCLNGKCKSYRLSYG
(SEQ ID NO: 162) Medicago truncatula >gi|152217940|gb|ABS31404.1| NCR 129 MQRKKNMGQILIFVFALINFLSPILVEMTTTTIPCTFIDDCPKMPLVVKCIDNFCNYFEIK
(SEQ ID NO: 163) Medicago truncatula >gi|152217938|gb|ABS31403.1| NCR 128 MAQTLMLVYALIIFTSLFLVVISRQTDIPCKSDDACPVSSHHIECVKGFCTYWKLD
(SEQ ID NO: 164) Medicago truncatula >gi|152217936|gb|ABS31402.1| NCR 127 MLRRKNTVQILMFVSALLIYIFLFLVITSSANIPCNSDSDCPWKIYYTYRCNDGFCVYKSIDPSTIPQYMTDLIFPR
(SEQ ID NO: 165) Medicago truncatula >gi|152217934|gb|ABS31401.1| NCR 122 MAVILKFVYIMIIFLFLLYVVNGTRCNRDEDCPFICTGPQIPKCVSHICFCLSSGKEAY
(SEQ ID NO: 166) Medicago truncatula >gi|152217932|gb|ABS31400.1| NCR 121 MDAILKFIYAMFLFLFLFVTTRNVEALFEECNRDFVCGNDDECVYPYAVQCIHRYCKCLKSRN
(SEQ ID NO: 167) Medicago truncatula >gi|152217930|gb|ABS31399.1| NCR 119 MQIGRKKMGETPKLVYVIILFLSIFLCTNSSFSQMINFRGCKRDKDCPQFRGVNIRCRSGFCTPIDS
(SEQ ID NO: 168) Medicago truncatula >gi|152217928|gb|ABS31398.1| NCR 118 MQMRKNMAQILFYVYALLILFSPFLVARIMVVNPNNPCVTDADCQRYRHKLATRMVCNIGFCLMDFTHDPYAPSLP
(SEQ ID NO: 169) Medicago truncatula >gi|152217926|gb|ABS31397.1| NCR 111 MYVYYIQMGKNMAQRFMFIYALIIFLSQFFVVINTSDIPNNSNRNSPKEDVFCNSNDDCPTTILYYVSKCVYNFCEYW
(SEQ ID NO: 170) Medicago truncatula >gi|152217924|gb|ABS31396.1| NCR 103 MAKIVNFVYSMIIFVSLFLVATKGGSKPFLTRPYPCNTGSDCPQNMCPPGYKPGCEDGYCNHCYKRW
(SEQ ID NO: 171) Medicago truncatula >gi|152217922|gb|ABS31395.1| NCR 101 MVRTLKFVYVIILILSLFLVAKGGGKKIYCENAASCPRLMYPLVYKCLDNKCVKFMMKSRFV
(SEQ ID NO: 172) Medicago truncatula >gi|152217920|gb|ABS31394.1| NCR96 MARTLKFVYAVILFLSLFLVAKGDDVKIKCVVAANCPDLMYPLVYKCLNGICVQFTLTFPFV
(SEQ ID NO: 173) Medicago truncatula >gi|152217918|gb|ABS31393.1| NCR94 MSNTLMFVITFIVLVTLFLGPKNVYAFQPCVTTADCMKTLKTDENIWYECINDFCIPFPIPKGRK
(SEQ ID NO: 174) Medicago truncatula >gi|152217916|gb|ABS31392.1| NCR93 MKRVVNMAKIVKYVYVIIIFLSLFLVATKIIEGYYYKCFKDSDCVKLLCRIPLRPKCMYRHICKCKVVLTQNNYVLT
(SEQ ID NO: 175) Medicago truncatula >gi|152217914|gb|ABS31391.1| NCR90 MKRGKNMSKILKFIYATLVLYLFLVVTKASDDECKIDGDCPISWQKFHTYKCINQKCKWVLRFHEY
(SEQ ID NO: 176) Medicago truncatula >gi|152217912|gb|ABS31390.1| NCR 88 MAKTLNFMFALILFISLFLVSKNVAIDIFVCQTDADCPKSELSMYTWKCIDNECNLFKVMQQMV
(SEQ ID NO: 177) Medicago truncatula >gi|152217910|gb|ABS31389.1| NCR 86 MANTHKLVSMILFIFLFLVANNVEGYVNCETDADCPPSTRVKRFKCVKGECRWTRMSYA
(SEQ ID NO: 178) Medicago truncatula >gi|152217908|gb|ABS31388.1| NCR 77 MAHFLMFVYALITCLSLFLVEMGHLSIHCVSVDDCPKVEKPITMKCINNYCKYFVDHKL
(SEQ ID NO: 179) Medicago truncatula >gi|152217906|gb|ABS31387.1| NCR 76 MNQIPMFGYTLIIFFSLFPVITNGDRIPCVTNGDCPVMRLPLYMRCITYSCELFFDGPNLCAVERI
(SEQ ID NO: 180) Medicago truncatula >gi|152217904|gb|ABS31386.1| NCR 74 MRKDMARISLFVYALIIFFSLFFVLTNGELEIRCVSDADCPLFPLPLHNRCIDDVCHLFTS
(SEQ ID NO: 181) Medicago truncatula >gi|152217902|gb|ABS31385.1| NCR68 MAQILMFVYFLIIFLSLFLVESIKIFTEHRCRTDADCPARELPEYLKCQGGMCRLLIKKD
(SEQ ID NO: 182) Medicago truncatula >gi|152217900|gb|ABS31384.1| NCR65 MARVISLFYALIIFLFLFLVATNGDLSPCLRSGDCSKDECPSHLVPKCIGLTCYCI
(SEQ ID NO: 183) Medicago truncatula >gi|152217898|gb|ABS31383.1| NCR62 MQRRKNMAQILLFAYVFIISISLFLVVTNGVKIPCVKDTDCPTLPCPLYSKCVDGFCKMLSI
(SEQ ID NO: 184) Medicago truncatula >gi|152217896|gb|ABS31382.1| NCR 57 MNHISKFVYALIIFLSVYLVVLDGRPVSCKDHYDCRRKVKIVGCIFPQEKPMCINSMCTCIREIVP
(SEQ ID NO: 185) Medicago truncatula >gi|152217894|gb|ABS31381.1| NCR 56 MKSQNHAKFISFYKNDLFKIFQNNDSHFKVFFALIIFLYTYLHVTNGVFVSCNSHIHCRVNNHKIGCNIPEQYLLCVNLFCLWLDY
(SEQ ID NO: 186) Medicago truncatula >gi|152217892|gb|ABS31380.1| NCR 54 MTYISKVVYALIIFLSIYVGVNDCMLVTCEDHFDCRQNVQQVGCSFREIPQCINSICKCMKG
(SEQ ID NO: 187) Medicago truncatula >gi|152217890|gb|ABS31379.1| NCR 53 MTTHISKFVFALIIFLSIYVGVNDCKRIPCKDNNDCNNNWQLLACRFEREVPRCINSICKCMPM
(SEQ ID NO: 188) Medicago truncatula >gi|152217888|gb|ABS31378.1| NCR 43 MVQTPKLVYVIVLLLSIFLGMTICNSSFSHFFEGACKSDKDCPKLHRSNVRCRKGQCVQI
(SEQ ID NO: 189) Medicago truncatula >gi|152217886|gb|ABS31377.1| NCR 28 MTKILMLFYAMIVFHSIFLVASYTDECSTDADCEYILCLFPIIKRCIHNHCKCVPMGSIEPMSTIPNGVHKFHIINN
(SEQ ID NO: 190) Medicago truncatula >gi|152217884|gb|ABS31376.1| NCR 26 MAKTLNFVCAMILFISLFLVSKNVALYIIECKTDADDCPISKLNMYNWRCIKSSCHLYKVIQFMV
(SEQ ID NO: 191) Medicago truncatula >gi|152217882|gb|ABS31375.1| NCR 24 MQKEKNMAKTFEFVYAMIIFILLFLVENNFAAYIIECQTDDDCPKSQLEMFAWKCVKNGCHLFGMYEDDDDP
(SEQ ID NO: 192) Medicago truncatula >gi|152217880|gb|ABS31374.1| NCR 21 MAATRKFIYVLSHFLFLFLVTKITDARVCKSDKDCKDIIIYRYILKCRNGECVKIKI
(SEQ ID NO: 193) Medicago truncatula >gi|152217878|gb|ABS31373.1| NCR 20 MQRLDNMAKNVKFIYVIILLLFFLLVIIVCDSAFVPNSGPCTTDKDCKQVKGYIARCRKGYCMQSVKRTWSSYSR
(SEQ ID NO: 194) Medicago truncatula >gi|152217876|gb|ABS31372.1| NCR 19 MKFIYIMILFLSLFLVQFLTCKGLTVPCENPTTCPEDFCTPPMITRCINFICLCDGPEYAEPEYDGPEPEYDHKGDFLSVKPKIINENMMMRERHMMKEIEV
(SEQ ID NO: 195) Medicago truncatula >gi|152217874|gb|ABS31371.1| NCR 12 MAQFLMFIYVLIIFLYLFYVEAAMFELTKSTIRCVTDADCPNVVKPLKPKCVDGFCEYT
(SEQ ID NO: 196) Medicago truncatula >gi|152217872|gb|ABS31370.1| NCR 10 MKMRIHMAQIIMFFYALIIFLSPFLVDRRSFPSSFVSPKSYTSEIPCKARDCPYELYYETKCVDSLCTY
(SEQ ID NO: 197) Medicago truncatula

Any NCR peptide known in the art is suitable for use in the methods or compositions described herein. Plants producing NCR peptides include, without limitationPisum sativum (peas),Astragalus sinicus (bean IRLC),Phaseolus vulgaris(beans),Vigna unguiculata (cowpeas)Medicago truncatula(truncated alfalfa) andLotus japonicus. For example, based on the genomic sequenceM. truncatula over 600 potential NCR peptides are predicted, and nearly 150 different NCR peptides have been detected in cells isolated from root nodules by mass spectrometry.

The NCR peptides described herein may be mature or immature NCR peptides. Immature NCR peptides have a C-terminal signal peptide that is required for translocation to the endoplasmic reticulum and is cleaved off after translocation. The N-terminus of the NCR peptide contains a signal peptide that can be cleaved to target the secretory pathway. NCR peptides are typically small peptides with disulfide bridges that stabilize their structures. Mature NCR peptides range in length from about 20 to about 60 amino acids, from about 25 to about 55 amino acids, from about 30 to about 50 amino acids, from about 35 to about 45 amino acids, or any range in between. NCR peptides may contain a conserved sequence of cysteine residues, while the remainder of the peptide sequence is highly variable. NCR peptides typically have approximately four or eight cysteine molecules.

NCR peptides can be anionic, neutral or cationic. In some instances, synthetic NCR cationic peptides having a pI greater than about eight have forms of antimicrobial activity. For example, both NCR247 (pI=10.15) (RNGCIVDPRCPYQQCRRPLYCRRR; SEQ ID NO: 198) and NCR335 (pI=11.22) (MAQFLLFVYSLIIFLSLFFGEAAFERTETRMLTIPCTSDDNCPKVISPCHTKCFDGFCGWYIEGSYEGP; SEQ ID NO: 199) are effective against Gram-negative and Gram-positive bacteria. also mushrooms. In some instances, neutral and/or anionic NCR peptides, such as NCR001, do not exhibit forms of antimicrobial activity at pI greater than about 8.

In some cases, the NCR peptide is effective in killing bacteria. In some cases, the NCR peptide is effective in killing S. meliloti, Xenorhabdus spp, Photorhabdus spp, Candidatus spp, Buchnera spp, Blattabacterium spp, Baumania spp, Wigglesworthia spp, Wolbachia spp, Rickettsia spp, Orientia spp, Sodalis spp, Burkholderia spp, Cupriavidus spp, Frankia spp, Snirhizobium spp, Streptococcus spp, Wolinella spp, Xylella spp, Erwinia spp, Agrobacterium spp, Bacillus spp, Paenibacillus spp, Streptomyces spp, Micrococcus spp, Corynebacterium spp, Acetobacter spp, Cyanobacteria spp, Rpphodocuscus, Pseudomonas spp, Lactobacillus spp, Enterococcus spp, Alcaligenes spp, Klebsiella spp, Paenibacillus spp, Arthrobacter spp, Corynebacterium spp, Brevibacterium spp, Thermus spp, Pseudomonas spp, Clostridium spp or Escherichia spp .

In some instances, the NCR peptide is a functionally active variant of the NCR peptide described herein. In some cases, the NCR peptide variant is at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical, for example, in a certain area or in the entire sequence, to the sequence of the NCR peptide described herein, or the NCR peptide of natural origin.

In some instances, the NCR peptide can be bioengineered to modulate its biological activity, eg increase or decrease or regulate, or to determine its target microorganism. In some cases, the NCR peptide is produced by the translation apparatus (eg, ribosome, etc.) of the cell. In some cases, the NCR peptide is chemically synthesized. In some cases, the NCR peptide is formed from a polypeptide precursor. The polypeptide precursor may be cleaved (eg, processed by a protease) to form the NCR peptide itself. Thus, in some cases, the NCR peptide is produced from a precursor polypeptide. In some instances, an NCR peptide includes a polypeptide that has undergone post-translational modifications, such as cleavage or the addition of one or more functional groups.

The NCR peptide described herein may be formulated for any of the uses described herein. The compositions disclosed herein may contain any number or type of NCR peptides, such as at least about any of 1 NCR peptide, 2, 3, 4, 5, 10, 15, 20, 30, 40, 50 , 100 or more NCR peptides. The appropriate concentration of each NCR peptide in the composition depends on such factors as potency, stability of the NCR peptide, amount of different NCR peptides, composition and methods of application of the composition. In some instances, the concentration of each NCR peptide in the liquid composition is from about 0.1 ng/mL to about 100 mg/mL. In some instances, the concentration of each NCR peptide in the solid composition is from about 0.1 ng/g to about 100 mg/g. In some cases, if the composition contains at least two types of NCR peptides, then the concentration of each type of NCR peptide may be the same or different.

A modulator comprising an NCR peptide described herein may be contacted with a target host in an amount and for a time sufficient to: (a) achieve a target level (e.g., a predetermined or threshold level) of peptide concentration NCR within the target host; (b) achieving a target level (eg, a predetermined or threshold level) for the concentration of the NCR peptide within the gut of the target host; (c) achieving a target level (eg, a predetermined or threshold level) of the NCR peptide concentration within the target host bacteriocyte; (d) modulating the level or activity of one or more microorganisms (eg, endosymbionts) in the target host or/and (e) modulating the fitness of the target host.

Regulatory peptides of bacteriocytes

The modulator described herein may include bacteriocyte regulatory peptide (BRP). BRPs are peptides expressed in insect bacteriocytes. These genes are first expressed at the time of development, coinciding with the inclusion of symbionts, and their bacteriocyte-specific expression is maintained throughout the life of the insect. In some cases, BRP has a hydrophobic amino-terminal domain, which is assumed to be a signal peptide. In addition, some BRPs have a cysteine-rich domain. In some instances, the bacteriocyte regulatory peptide is a bacteriocyte-specific cysteine-rich protein (BCR). Bacteriocyte regulatory peptides are about 40 to 150 amino acids in length. In some instances, the bacteriocyte regulatory peptide has a length in the range of about 45 to about 145, about 50 to about 140, about 55 to about 135, about 60 to about 130, about 65 to about 125, about 70 to about 120, about 75 to about 115, about 80 to about 110, about 85 to about 105, or any range in between. Non-limiting examples of BRPs and their forms of activity are shown in Table 8.

Table 8. Examples of bacteriocyte regulatory peptides

Name Peptide sequence Cysteine-rich bacteriocyte-specific BCR protein family, BCR1 peptide MKLLHGFLIIMLTMHLSIQYAYGGPFLTKYLCDRVCHKLCGDEFVCSCIQYKSLKGLWFPHCPTGKASVVLHNFLTSP
(SEQ ID NO: 200) Cysteine-rich bacteriocyte-specific BCR protein family, BCR2 peptide MKLLYGFLIIMLTIHLSVQYFESPFETKYNCDTHCNKLCGKIDHCSCIQYHSMEGLWFPHCRTGSAAQMLHDFLSNP
(SEQ ID NO: 201) Cysteine-rich bacteriocyte-specific BCR protein family, BCR3 peptide MSVRKNVLPTMFVVLLIMSPVTPTSVFISAVCYSGCGSLALVCFVSNGITNGLDYFKSAPLSTSETSCGEAFDTCTDHCLANFKF
(SEQ ID NO: 202) Cysteine-rich bacteriocyte-specific BCR protein family, BCR4 peptide MRLLYGFLIIMLTIYLSVQDFDPTEFKGPFPTIEICSKYCAVVCNYTSRPCYCVEAAKERDQWFPYCYD
(SEQ ID NO: 203) Cysteine-rich bacteriocyte-specific BCR protein family, BCR5 peptide MRLLYGFLIIMLTIHLSVQDIDPNTLRGPYPTKEICSKYCEYNVVCGASLPCICVQDARQLDHWFACCYDGGPEMLM
(SEQ ID NO: 204) SP family of secreted proteins, SP1 peptide MKLFVVVVLVAVGIMFVFASDTAAAPTDYEDTNDMISLSSLVGDNSPYVRVSSADSGGSSKTSSKNPILGLLKSVIKLLTKIFGTYSDAAPAMPPIPPALRKNRGMLA
(SEQ ID NO: 205) SP family of secreted proteins, SP2 peptide MVACKVILAVAVVFVAAVQGRPGGEPEWAAPIFAELKSVSDNITNLVGLDNAGEYATAAKNNLNAFAESLKTEAAVFSKSFEGKASASDVFKESTKNFQAVVDTYIKNLPKDLTLKDFTEKSEQALKYMVEHGTEITKKAQGNTETEKEIKEFFKKQIENLIGQGKALQAKIAEAKKA
(SEQ ID NO: 206) SP family of secreted proteins, SP3 peptide MKTSSSKVFASCVAIVCLASVANALPVQKSVAATTENPIVEKHGCRAHKNLVRQNVVDLKTYDSMLITNEVVQKQSNEVQSSEQSNEGQNSEQSNEGQNSEQSNEVQSSEHSNEGQNSKQSNEGQNSEQSNEVQSSEHSNEGQNSEQSNEVQSSEHSNEGQNSKQSNEGQNSKQSNEVQSSEHWNEGQNSKQSNEDQNSEQSNEGQNSKQSNEGQNSKQSNEDQNSEQSNEGQNSKQSNEVQSSEQSNEGQNSKQSNEGQSSEQSNEGQNSKQSNEVQSPEEHYDLPDPESSYESEETKGSHESGDDSEHR
(SEQ ID NO: 207) SP family of secreted proteins, SP4 peptide MKTIILGLCLFGALFWSTQSMPVGEVAPAVPAVPSEAVPQKQVEAKPETNAASPVSDAKPESDSKPVDAEVKPTVSEVKAESEQKPSGEPKPESDAKPVVASESKPESDPKPAAVVESKPENDAVAPETNNDAKPENAAAPVSENKPATDAKAETELIAQAKPESKPASDLKAEPEAAKPNSEVPVALPLNPTETKATQQSVETNQVEQAAPAAAQADPAAAPAADPAPAPAAAPVAAEEAKLSESAPSTENKAAEEPSKPAEQQSAKPVEDAVPAASEISETKVSPAVPAVPEVPASPSAPAVADPVSAPEAEKNAEPAKAANSAEPAVQSEAKPAEDIQKSGAVVSAENPKPVEEQKPAEVAKPAEQSKSEAPAEAPKPTEQSAAEEPKKPESANDEKKEQHSVNKRDATKEKKPTDSIMKKQKQKKAN
(SEQ ID NO: 208) SP secreted protein family, SP5a peptide MNGKIVLCFAVVFIGQAMSAATGTTPEVEDIKKVAEQMSQTFMSVANHLVGITPNSADAQKSIEKIRTIMNKGFTDMETEANKMKDIVRKNADPKLVEKYDELEKELKKHLSTAKDMFEDKVVKPIGEKVELKKITENVIKTTKDMEATMNKAIDGFKKQ
(SEQ ID NO: 209) SP family of secreted proteins, SP6 peptide MHLFLALGLFIVCGMVDATFYNPRSQTFNQLMERRQRSIPIPYSYGYHYNPIEPSINVLDSLSEGLDSRINTFKPIYQNVKMSTQDVNSVPRTQYQPKNSLYDSEYISAKDIPSLFPEEDSYDYKYLGSPLNKYLTRPSTQESGIAINLVAIKETSVFDYGFPTYKSPYSSDSVWNFGSKIPNTVFEDPQSVESDPNTFKVSSPTIKIVKLLPETPEQESIITTTKNYELNYKTTQETPTEAELYPITSEEFQTEDEWHPMVPKENTTKDESSFITTEEPLTEDKSNSITIEKTQTEDESNSIEFNSIRTEEKSNSITTEENQKEDDESMSTTSQETTTAFNLNDTFDTNRYSSSHESLMLRIRELMKNIADQQNKSQFRTVDNIPAKSQSNLSSDESTNQQFEPQLVNGADTYK
(SEQ ID NO: 210) Coloptericin A, ColA peptide MTRTMLFLACVAALYVCISATAGKPEEFAKLSDEAPSNDQAMYESIQRYRRFVDGNRYNGGQQQQQQPKQWEVRPDLSRDQRGNTKAQVEINKKGDNHDINAGWGKNINGPDSHKDTWHVGGSVRW
(SEQ ID NO: 211) RlpA type I MKETTVVWAKLFLILIILAKPLGLKAVNECKRLGNNSCRSHGECCSGFCFIEPGWALGVCKRLGTPKKSDDSNNGKNIEKNNGVHERIDDVFERGVCSYYKGPSITANGDVFDENEMTAAHRTLPFNTMVKVEGMGTSVVVKINDRKTAADGKVMLLSRAAAESLNIDENTGPVQCFCQLKFVLDGSGCTPDYGDTCVLHHECCSQGLPSEK21

In some cases, BRP alters the growth and/or activity of one or more bacteria residing in the host bacteriocyte. In some cases, BRP can be created using bioengineering to modulate its biological activity (for example, increase, decrease or regulate) or to determine its target microorganism. In some cases, BRP is produced by the translation apparatus (eg, ribosome, etc.) of the cell. In some cases, BRP is synthesized chemically. In some cases, BRP is formed from a polypeptide precursor. The precursor polypeptide may be cleaved (eg, processed by a protease) to form the BRP polypeptide itself. Thus, in some cases, BRP is produced from a precursor polypeptide. In some instances, the BRP includes a polypeptide that has undergone post-translational modifications, such as cleavage or the addition of one or more functional groups.

The functionally active BRP variants described herein are also useful in the compositions and methods described herein. In some cases, the BRP variant is at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83 %, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical , for example, in a certain area or in the entire sequence, the sequence of the BRP described in this document, or BRP of natural origin.

The BRP described herein may be formulated for any of the uses described herein. The compositions disclosed herein may contain any number or type (eg, classes) of BRPs, such as at least about any of 1 BRP, 2, 3, 4, 5, 10, 15, 20 or more BRPs. . The appropriate concentration of each BRP in the composition depends on such factors as the potency, stability of the BRP, the amount of different BRPs, composition and methods of application of the composition. In some instances, the concentration of each BRP in the liquid composition is from about 0.1 ng/mL to about 100 mg/mL. In some cases, the concentration of each BRP in the solid composition is from about 0.1 ng/g to about 100 mg/g. In some cases, if the composition contains at least two types of BRP, then the concentration of each type of BRP may be the same or different.

A modulator comprising a BRP described herein may be contacted with a target host in an amount and for a time sufficient to: (a) achieve a target level (e.g., a predetermined or threshold level) of the BRP concentration host-target; (b) achieving a target level (eg, a predetermined or threshold level) of BRP concentration within the gut of the target host; (c) achieving a target level (eg, a predetermined or threshold level) of BRP concentration within the target host bacteriocyte; (d) modulating the level or activity of one or more microorganisms (eg, endosymbionts) in the target host or/and (e) modulating the fitness of the target host.

small molecules

Numerous small molecules (eg, antibiotic or metabolite) may be useful in the compositions and methods described herein. In some cases, an effective concentration of any small molecule described herein can alter the level, activity or metabolism of one or more microorganisms (described above) living in the host, the change resulting in increased host fitness.

A modulator comprising a small molecule as described herein may be contacted with a target host in an amount and for a time sufficient to: (a) achieve a target level (e.g., a predetermined or threshold level) of a small concentration molecules within the target host; (b) achieving a target level (eg, a predetermined or threshold level) of the concentration of a small molecule within the gut of the target host; (c) achieving a target level (eg, a predetermined or threshold level) of the small molecule concentration within the target host bacteriocyte; (d) modulating the level or activity of one or more microorganisms (eg, endosymbionts) in the target host or/and (e) modulating the fitness of the target host.

Antibiotics

The modulator described herein may include an antibiotic. Any antibiotic known in the art can be used. Antibiotics are usually classified based on their mechanism of action, chemical structure, or spectrum of activity.

The antibiotic described herein can target any function or process of bacterial growth and can be either bacteriostatic (eg, inhibit or prevent bacterial growth) or bactericidal (eg, kill bacteria). In some cases, the antibiotic is a bactericidal antibiotic. In some cases, a bactericidal antibiotic is an antibiotic that targets the bacterial cell wall (eg, penicillins and cephalosporins); an antibiotic that targets the cell membrane (for example, polymyxins); or an antibiotic that inhibits essential bacterial enzymes (eg, rifamycins, lipiarmycins, quinolones, and sulfonamides). In some cases, the bactericidal antibiotic is an aminoglycoside. In some cases, the antibiotic is a bacteriostatic antibiotic. In some cases, a bacteriostatic antibiotic specifically affects protein synthesis (eg, macrolides, lincosamides, and tetracyclines). Additional classes of antibiotics that may be used herein include cyclic lipopeptides (such as daptomycin), glycylcyclines (such as tigecycline), oxazolidinones (such as linezolid), or lipiaarmycins (such as fidaxomycin). Examples of antibiotics include oxytetracycline, doxycycline, rifampicin, ciprofloxacin, ampicillin, and polymyxin B. Other non-limiting examples of antibiotics are in Table 9.

Table 9 Examples of antibiotics

Antibiotics Action Penicillins, cephalosporins, vancomycin Cell wall synthesis polymyxin, gramicidin Membrane-active agent, destroys the cell membrane Tetracyclines, macrolides, chloramphenicol, clindamycin, spectinomycin Inhibit protein synthesis Sulfonamides Inhibit folate-dependent pathways Ciprofloxacin Inhibit DNA gyrase Isoniazid, rifampicin, pyrazinamide, ethambutol, (miambutol), streptomycin Antimycobacterial agents

An antibiotic described herein may have any level of target specificity (eg, narrow or broad spectrum). In some cases, the antibiotic is a narrow spectrum antibiotic and thus targets specific types of bacteria, such as Gram-negative or Gram-positive bacteria. Alternatively, the antibiotic may be a broad spectrum antibiotic that targets a broad range of bacteria.

The antibiotics described herein may be formulated for any of the routes of administration described herein. The compositions disclosed herein may contain any number or type (e.g., classes) of antibiotics, such as at least about any of 1 antibiotic, 2, 3, 4, 5, 10, 15, 20 or more antibiotics. (for example, a combination of rifampicin and doxycycline, or a combination of ampicillin and rifampicin). The appropriate concentration of each antibiotic in the composition depends on factors such as potency, stability of the antibiotic, amount of different antibiotics, composition and methods of application of the composition. In some cases, if the composition contains at least two types of antibiotics, then the concentration of each type of antibiotic may be the same or different.

A modulator comprising an antibiotic as described herein may be contacted with a target host in an amount and for a time sufficient to: (a) achieve a target level (e.g., a predetermined or threshold level) of the antibiotic concentration within host-target; (b) reaching a target level (eg, a predetermined or threshold level) of the antibiotic concentration within the gut of the target host; (c) achieving a target level (eg, a predetermined or threshold level) of the antibiotic concentration within the target host bacteriocyte; (d) modulating the level or activity of one or more microorganisms (eg, endosymbionts) in the target host or/and (e) modulating the fitness of the target host.

Secondary metabolites

In some instances, the modulating agent in the compositions and methods described herein includes a secondary metabolite. Secondary metabolites are formed from organic molecules produced by the body. Secondary metabolites can act as (i) competitive agents against bacteria, fungi, amoebae, plants, insects and large animals; (ii) as a means of transporting metals; (iii) as a means of symbiosis between microbes and plants, insects and larger animals; (iv) as sex hormones; and (v) as differentiation effectors. Non-limiting examples of secondary metabolites are in Table 10.

Table 10. Examples of secondary metabolites

Phenylpropanoids alkaloids Terpenoids Quinones Steroids Polyketides Anthocyanins Acridines Carotenes Anthroquinones cardiac Erythromycin Coumarins Betalains Monoterpenes Benzoquinones Glycosides Lovastatin and other statins Flavonoids Quinolizidins Sesquiterpenes Naphthoquinones Derivatives Discodermolide Derivatives Furonoquinones Diterpenes pregnenolone Aflatoxin B1 hydroxycinnamoyl harringtonins Triterpenes Avermectins Isoflavones isoquinolines Nystatin Lignans indoles Rifamycin Phenolenones Purines Proanthocyanidins Pyridines Stilbens tropane Tannins alkaloids

The secondary metabolite used herein may include a metabolite from any known group of secondary metabolites. For example, secondary metabolites can be classified into the following groups: alkaloids, terpenoids, flavonoids, glycosides, natural phenols (e.g. gossypolyacetic acid), enals (e.g. trans-cinnamic aldehyde), phenazines, biphenols and dibenzofurans, polyketides, fatty acid synthase peptides, non-ribosomal peptides, peptides synthesized in ribosomes and subject to post-translational modifications, polyphenols, polysaccharides (eg chitosan) and biopolymers. For an in-depth review of secondary metabolites see, for example, Vining, Annu. Rev. microbiol. 44:395-427, 1990. Secondary metabolites useful in the compositions and methods described herein include metabolites that alter the natural function of an endosymbiont (eg, primary or secondary endosymbiont), bacteriocyte, or extracellular symbiont. In some instances, one or more secondary metabolites described herein are isolated from high throughput screening (HTS) of antimicrobial compounds. For example, HTS screening identified 49 antibacterial extracts that are specific for Gram-positive and Gram-negative bacteria from over 39,000 crude extracts from organisms growing in various ecosystems in one particular region. In some cases, the secondary metabolite is transported into the bacteriocyte.

In some cases, the small molecule is an inhibitor of vitamin synthesis. In some cases, the vitamin synthesis inhibitor is an analog of a vitamin precursor. In certain instances, the precursor analog of the vitamin is pantothenol.

In some cases, the small molecule is an amino acid analogue. In certain instances, the amino acid analog is L-canavanine, D-arginine, D-valine, D-methionine, D-phenylalanine, D-histidine, D-tryptophan, D-threonine, D-leucine, L-NG-nitroarginine, or their combination.

In some cases, the small molecule is a natural antimicrobial compound such as propionic acid, levulinic acid, trans-cinnamic aldehyde, nisin, or low molecular weight chitosan.

The secondary metabolite described herein may be formulated for any of the routes of administration described herein. The compositions disclosed herein may contain any number or type (eg, classes) of secondary metabolites, such as at least about any of 1 secondary metabolite, 2, 3, 4, 5, 10, 15, 20 or more secondary metabolites. The appropriate concentration of each secondary metabolite in the composition depends on such factors as potency, stability of the secondary metabolite, amount of various secondary metabolites, composition and methods of application of the composition. In some cases, if the composition contains at least two types of secondary metabolites, then the concentration of each type of secondary metabolite may be the same or different.

A modulating agent comprising a secondary metabolite described herein may be contacted with a target host in an amount and for a time sufficient to: (a) achieve a target level (e.g., a predetermined or threshold level) of the secondary metabolite within the target host; (b) achieving a target level (eg, a predetermined or threshold level) of the secondary metabolite concentration within the gut of the target host; (c) reaching a target level (eg, a predetermined or threshold level) of the secondary metabolite concentration within the target host bacteriocyte; (d) modulating the level or activity of one or more microorganisms (eg, endosymbionts) in the target host or/and (e) modulating the fitness of the target host.

Bacteria as modulators

In some cases, the modulator described herein includes one or more bacteria. Numerous bacteria are useful in the compositions and methods described herein. In some cases, the agent is a bacterial species found endogenously in the host. In some instances, the bacterial modulator is a species of endosymbiotic bacteria. Non-limiting examples of bacteria that can be used as modulators include all of the bacterial species described herein in Section II of the Detailed Description and the bacterial species listed in Table 1. For example, the modulator may be a bacterial species from any type bacteria present in insect gut and/or hemocoel, including Gammaproteobacteria, Alphaproteobacteria, Betaproteobacteria, Bacteroidetes, Firmicutes (e.g. Lactobacillus and Bacillus spp.), Clostridia, Actinomycetes, Spirochetes, Verrucomicrobia and Actinobacteria .

In some cases, the modulating agent is a bacterium that promotes microbial diversity or otherwise alters the host microbiota in a beneficial manner. In one case, a bacterium may be provided to promote the development of microbiota in insects.

The bacterial modulators discussed herein can be used to alter the level, activity, or metabolism of target micro-organisms, as outlined in the fitness-enhancing sections of insects such as crickets, grasshoppers, or locusts.

In some instances, such bacterial modulators are bacteria that are capable of producing nutrients, including amino acids (eg, methionine). The nutrient-producing bacteria may be naturally occurring bacteria, for example, naturally occurring bacteria exogenous to the insect host. Such bacteria can be isolated from a population of bacteria, such as the population found in a media sample. Bacteria can be isolated that produce one or more amino acids in such a way that the amino acid production of the host is increased relative to a host that has not been injected with the amino acid producing bacteria. Amino acids that can be produced by bacteria in the host include methionine, alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glutamate, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine , tryptophan, tyrosine or valine. In some cases, the amino acid producing bacteria are methionine producing bacteria.

In some instances, nutrient-producing bacteria (e.g., amino acid-producing bacteria, e.g., methionine-producing bacteria) are present at a concentration of at least 100,000 cells/mL (e.g., at least about 100,000 cells/mL, at least about 150,000 cells/mL, at least about 200,000 cells/mL, at least about 250,000 cells/mL, at least about 300,000 cells/mL, at least about 350,000 cells/mL, at least about 400,000 cells/mL , at least about 450,000 cells/ml or at least about 500,000 cells/ml).

Examples 1-4 and 8 describe how methionine-producing microorganisms can be identified, which can then be used as modulators in insect hosts such as crickets or in the Drosophila model organism to increase host fitness (e.g., increase amino acid content ( e.g. methionine content)). For example, in some cases, the nutrient content is increased in the host prior to the host's use in the manufacture of food or feed.

In some instances, such bacterial modulators are bacteria that are capable of degrading pesticides as set forth in Table 12, including insecticides. Such insecticides include neonicotinoids such as imidacloprid or organophosphate insecticides such as fenitrothion. In some instances, pesticide-metabolizing bacteria are present at a concentration of at least 100,000 cells/mL (e.g., at least about 100,000 cells/mL, at least about 150,000 cells/mL, at least about 200,000 cells/mL, at least about 250,000 cells/mL, at least about 300,000 cells/mL, at least about 350,000 cells/mL, at least about 400,000 cells/mL, at least about 450,000 cells/mL, or at least about 500,000 cells/mL ).

Examples 5 and 6 describe how imidacloprid and fenitrothion degrading microorganisms can be identified, which can then be used as modulators in insect hosts such as crickets, providing treated insect hosts with a competitive advantage. Administration of such pesticide degrading microorganisms, eg, imidacloprid or fenitrothion degrading microorganisms, to insect hosts such as honey bees is understood to involve altering the level, activity, or metabolism of one or more microorganisms living in the host.

Modifications regarding modulating means

Merger Products

Any of the modulators described herein may be fused or associated with an additional fragment. In some cases, the modulator includes a fusion product of one or more additional fragments (eg, 1 additional fragment, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more additional fragments). In some instances, the additional moiety is any of the modulators described herein (eg, a peptide, polypeptide, small molecule, or antibiotic). Alternatively, the additional fragment may not itself act as a modulator, but may instead serve a secondary function. For example, the additional moiety may facilitate access, binding, or activation of the modulator at a target site in the host (eg, host gut or bacteriocyte) or in a target microorganism in the host (eg, cricket, grasshopper, or locust).

In some instances, the additional moiety may facilitate entry of the modulating agent into the target host cell or target microorganism within the host. For example, the additional fragment may include a cell-penetrating peptide. Cell penetrating peptides (CPPs) may be natural sequences derived from proteins; chimeric peptides, which are formed by the fusion of two natural sequences; or synthetic CPPs, which are synthetically engineered sequences based on structure-activity relationship studies. In some cases, CPPs are characterized by the ability to ubiquitously penetrate cell membranes (eg, prokaryotic and eukaryotic cell membranes) with limited toxicity. In addition, CPPs may be characterized by the ability to cross cell membranes through volatile and/or nonvolatile mechanisms without the need for chiral recognition by specific receptors. CPPs can communicate with any of the modulators described in this document. For example, CPP can bind to an antimicrobial peptide (AMP), eg, a scorpion peptide, eg, UY192, fused to a cell-penetrating peptide (eg, YGRKKRRQRRRFLSTIWNGIKGLLFAM; SEQ ID NO: 226). Non-limiting examples of CPPs are shown in Table 11.

Table 11 Examples of Cell Penetrating Peptides (CPPs)

Peptide Origin Subsequence Derived from proteins Penetratin antennapedia RQIKIWFQNRRMKWKK
(SEQ ID NO: 213) Peptide Tat Tat GRKKRRQRRRPPQ
(SEQ ID NO: 214) pVEC Cadherin LLIILRRRIRKQAHAHSK
(SEQ ID NO: 215) Chimera Transportan Galanin/Mastoparan GWTLNSAGYLLGKINLKALAALAKKIL
(SEQ ID NO: 216) MPG HIV-gp41/T-antigen SV40 GALFLGFLGAAGSTMGAWSQPKKKRKV
(SEQ ID NO: 217) Pep-1 HIV reverse transcriptase/SV40 T-antigen KETWWETWWTEWSQPKKKRKV
(SEQ ID NO: 218) Synthetic Polyarginines Based on Tat peptide (R) _n ; 6 <n < 12 MAP de novo KLALKLALKALKAALKLA
(SEQ ID NO: 219) R ₆ W ₃ based on penetratin RRWWRRWRR
(SEQ ID NO: 220)

In other instances, the additional moiety facilitates binding of the modulator to a target microorganism (eg, fungus or bacteria) residing in the host. The additional fragment may include one or more targeting domains. In some instances, the targeting domain may target the modulator to one or more microorganisms (eg, bacteria or fungus) living in the gut of the host. In some cases, the targeting domain may target the modulator to a specific area in the host body (eg, the gut or bacteriocyte of the host) to access microorganisms that are normally present in said area of the host body. For example, the targeting domain may target the modulator to the foregut, midgut, or hindgut of the host. In other cases, the targeting domain may target the modulator to a bacteriocyte in the host and/or one or more bacteria living in the bacteriocyte of the host.

Pre- or pro-domains

In some cases, the modulating agent may include an amino acid pre- and prosequence. For example, a modulating agent may be an inactive protein or peptide that may be activated by cleavage or post-translational modification of a pre- or pro-sequence. In some cases, the modulating agent is designed with an inactivating pre or pro sequence. For example, the pre- or prosequence may close an activation site on the modulator, such as a receptor binding site, or may induce a conformational change in the modulator. Thus, upon cleavage of the pre- or pro-sequence, the modulator is activated.

Alternatively, the modulating agent may include a small pre- or pro-molecule, such as an antibiotic. A modulating agent may be an inactive small molecule as described herein that can be activated in a target environment within a host. For example, a small molecule can be activated when a certain pH is reached in the gut of the host. For example, the targeting domain may be a Galanthus nivalis lectin or agglutinin (GNA) associated with a modulator described herein, eg AMP, eg a scorpion peptide, eg Uy192.

Linkers

In cases where the modulating agent is associated with an additional fragment, the modulating agent may additionally contain a linker. For example, the linker may be a chemical bond, such as one or more covalent bonds or non-covalent bonds. In some instances, the linker may be a peptide linker (eg, 2, 3, 4, 5, 6, 8, 10, 12, 14, 16, 20, 25, 30, 35, 40 or more amino acids long). The linker may include any of the flexible, rigid, or cleavable linkers described herein.

The flexible peptide linker may include any of the linkers commonly used in the art, including linkers having sequences containing primarily Gly and Ser residues ("GS" linker). Flexible linkers may be useful for connecting domains that require a certain degree of mobility or interaction, and may contain small, non-polar (eg, Gly) or polar (eg, Ser or Thr) amino acids.

Alternatively, the peptide linker may be a rigid linker. Rigid linkers are suitable for maintaining a fixed distance between fragments and maintaining their independent functions. Rigid linkers may also be suitable if the spatial separation of the domains is important to maintain the stability or biological activity of one or more components in the fusion product. Rigid linkers, for example, may have an alpha-helical structure or a Pro-rich sequence, (XP) _n , where X is any amino acid, preferably Ala, Lys or Glu.

In a few more instances, the peptide linker may be a cleavable linker. In some cases, linkers can be cleaved under specific conditions such as the presence of reducing agents or proteases. In vivo cleavable linkers can utilize the reversible nature of the disulfide bond. One example includes a thrombin sensitive sequence (eg, PRS) between two Cys residues . Thrombin treatment of CPRSC in vitro results in cleavage of the thrombin-responsive sequence, while the reversible disulfide bond remains intact. Such linkers are known and described, for example, in Chen et al., Adv. drug deliv. Rev. 65(10):1357-1369, 2013. Cleavage of linkers in fusion products can also be carried out by proteases that are expressed in vivo in specific cells or tissues of the host or microorganisms living in the host organism. In some cases, upon reaching the target site or cell, cleavage of the linker may release a free functional modulator.

The fusion products described herein may alternatively be linked to a linking molecule, including a hydrophobic linker such as a negatively charged sulfonate group; lipids such as poly(--CH2--) hydrocarbon chains such as a polyethylene glycol (PEG) group, their unsaturated variants, their hydroxylated variants, amidated or otherwise N-containing variants, non-carbon linkers; carbohydrate linkers; phosphodiester linkers, or other molecules capable of covalently linking two or more molecules, such as modulators. Non-covalent linkers can be used, such as hydrophobic lipid globules to which the modulating agent binds, for example via a hydrophobic region of the modulating agent or a hydrophobic extension of the modulating agent, such as a series of residues rich in leucine, isoleucine, valine, or possibly also alanine, phenylalanine, or even tyrosine, methionine, glycine or other hydrophobic moiety. The modulator may be coupled by charge chemistry such that a positively charged moiety of the modulator is associated with a negatively charged other modulator or additional moiety.

Compositions and compositions

The compositions described herein may be formulated in pure form (eg, the composition contains a modulating agent) or together with one or more additional agents (such as a vehicle, delivery vehicle, carrier, diluent, stabilizer, etc.) to facilitate application or delivery of the compositions. Examples of suitable fillers and diluents include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, starches, gum arabic, calcium phosphate, alginates, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, saline, syrup , methyl cellulose, meth and propyl hydroxybenzoates, talc, magnesium stearate and mineral oil.

In some cases, the composition contains a delivery device or carrier. In some cases, the delivery vehicle includes a filler. Illustrative excipients include, without limitation, solid or liquid carrier materials, solvents, stabilizers, excipients for sustained release formulations, colorants, and surfactants (surfactants). In some cases, the delivery vehicle is a stabilizing delivery vehicle. In some instances, the stabilizing delivery vehicle includes a stabilizing excipient. Exemplary stabilizing excipients include, but are not limited to, epoxidized vegetable oils, antifoam agents such as silicone oil, preservatives, viscosity regulators, binders, and tackifiers. In some cases, the stabilizing delivery agent is a buffer suitable for the modulating agent. In some instances, the composition is microencapsulated in a polymer bead-based delivery vehicle. In some cases, the stabilizing delivery agent protects the modulating agent from UV and/or acidic conditions. In some cases, the delivery vehicle contains a pH buffer. In some cases, the composition is formulated such that it has a pH in the range of from about 4.5 to about 9.0, including, for example, any of the pH ranges from about 5.0 to about 8.0, from about 6.5 to about 7.5, or about 6.5 to about 7.0.

Depending on the intended purpose and the prevailing circumstances, the composition may be formulated as emulsion concentrates, suspension concentrates, directly sprayable or dilute solutions, coating pastes, dilute emulsions, sprayable powders, soluble powders, dispersible powders, wettable powders, dust formulations. , granules, polymer encapsulated forms, microcapsules, foams, aerosols, carbon dioxide preparations, tablets, resin preparations, paper preparations, non-woven fabric preparations or knitted or woven fabric preparations. In some cases, the composition is a liquid. In some cases, the composition is a solid. In some cases, the composition is an aerosol, such as in a pressurized aerosol can. In some cases, the composition is present in the waste (such as excrement) of the pest. In some cases, the composition is present inside or on the surface of the body of a living pest.

In some cases, the delivery vehicle is food or water for the host. In other cases, the delivery vehicle is a food source for the host. In some cases, the delivery vehicle is a food bait for the host. In some instances, the composition is an edible that is ingested by the host. In some cases, the composition is delivered by the host to the second host and taken up by the second host. In some cases, the composition is taken up by the host or second host and the composition is released into the environment of the host or second host in the waste (such as feces) of the host or second host. In some cases, the modulating agent is included in a food bait intended to be taken up by the host or carried back to its colony.

In some cases, the modulating agent may comprise from about 0.1% to about 100% of the composition, such as any amount from about 0.01% to about 100%, from about 1% to about 99.9%, from about 0.1% to about 10%, about 1% to about 25%, about 10% to about 50%, about 50% to about 99%, or about 0.1% to about 90% of active ingredients (such as phage, lysine or bacteriocin). In some cases, the composition contains at least any of 0.1%, 0.5%, 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% , 90%, 95% or more active ingredients (such as phage, lysine and bacteriocin). In some cases, concentrated products are preferred as commercial products, and the end user typically uses diluted products that have a significantly lower concentration of the active ingredient.

Any of the formulations described herein may be applied in the form of a bait, coil, electric mat, fuming preparation, fumigant, or sheet.

Liquid formulations

The compositions provided herein may be in the form of a liquid formulation. Liquid formulations are usually mixed with water, however, in some cases they can be used with vegetable oil, diesel fuel, kerosene or other light oil as a carrier. The amount of active ingredient is often in the range of from about 0.5 to about 80 percent by weight.

An emulsion concentrate formulation may contain a liquid active ingredient, one or more petroleum-based solvents, and an agent that promotes mixing of the formulation with water to form an emulsion. Such concentrates can be used in formulations for agriculture, ornamental and lawn plants, forestry, building treatment, food processing, farm animals and public health pests. They can be adapted to field equipment from small portable sprayers to hydraulic sprayers, low volume ground sprayers, fan foggers and low volume aerial sprayers. Some active ingredients are readily soluble in the liquid carrier. When mixed with a carrier, they form a solution in which precipitation and separation do not occur, eg a homogeneous solution. Formulations of this type may contain an active ingredient, a carrier and one or more other ingredients. Solutions can be used in any type of sprayer, indoors and outdoors.

In some cases, the composition may be formulated as an inverse emulsion. An inverse emulsion is a water-soluble active ingredient dispersed in an oil carrier. Inverse emulsions require an emulsifier that promotes mixing of the active ingredient with a large volume of a petroleum-based carrier, typically a fuel oil. Reverse emulsions help reduce drift. With other formulations, some spray drift occurs as the water droplets begin to evaporate before reaching the target surfaces; as a result, the droplets can become very small and lightweight. Because oil evaporates more slowly than water, the inverse emulsion droplets shrink less and more of the active ingredient reaches its target. The oil additionally contributes to the reduction of surface runoff and increases resistance to rain. It additionally acts as an adhesive by improving surface coverage and absorption. Because the droplets are relatively large and heavy, it is difficult for them to pass through the underside cover of the leaves. Inverse emulsions are most widely used in right-of-way areas where drift into susceptible off-target areas can be a problem.

A flowable or liquid formulation combines many of the characteristics of emulsion concentrates and wettable powders. Manufacturers use these formulations if the active ingredient is a solid that is neither water nor oil soluble. The active ingredient impregnated with a substance such as clay is ground to a very fine powder. The powder is then suspended in a small amount of liquid. The resulting liquid product is quite dense. Flowable and liquid formulations have many of the characteristics of emulsion concentrates, and they also have similar disadvantages. They require moderate agitation to keep them in suspension and leave visible residues similar to those of wettable powders.

Flowable/liquid formulations are easy to handle and apply. Since they are liquids, they are subject to splashing and splashing. They contain solid particles, so they contribute to abrasive wear of nozzles and pumps. Flowable and liquid suspensions settle in their containers. Since flowable and liquid formulations tend to settle out, packaging in five gallon or smaller containers facilitates remixing.

Aerosol formulations contain one or more active ingredients and a solvent. Most aerosols have a low percentage of active ingredients. There are two types of aerosol formulations - the ready-to-use type, widely available in pressurized sealed containers, and products used in electric or gasoline-powered aerosol generators that release the formulation as smoke or mist.

Ready-to-use aerosol formulations are typically small, self-contained units that release the formulation when the nozzle valve is actuated. The composition is passed through a thin hole using an inert gas under pressure to form fine droplets. These products are used in greenhouses, small areas inside buildings or limited outdoor areas. Commercial models that hold up to 5 pounds of active ingredient are usually refillable.

Smoke-forming or fogging aerosol compositions are not pressurized. They are used in machines that break up a liquid formulation into a fine mist or thick mist (aerosol) using a rapidly rotating disk or heated surface.

Dry or solid formulations

Dry formulations can be divided into two types: ready-to-use and concentrates, which must be mixed with water to be applied as a spray. Most pulverulent formulations are ready to use and have a low percentage of active ingredients (less than about 10 percent by weight) along with a very fine dry inert carrier made from talc, chalk, clay, walnut shell or volcanic ash. The size of individual dust particles varies. Some pulverized formulations are concentrates and have a high percentage of active ingredients. Before use, they are mixed with dry inert carriers. Powders are always used dry and can easily be carried to non-target areas.

Granular or pellet formulations

In some cases, the composition is in the form of granules. Granular formulations are similar to dust formulations, except that the granular particles are larger and heavier. Coarse particles can be obtained from materials such as clay, corn cobs or walnut shells. The active ingredient coats the outer surface of the granules or is absorbed into them. The amount of active ingredient may be relatively small, typically in the range of from about 0.5 to about 15 percent by weight. Granular formulations are most commonly used for soil application, soil insects, or absorption by plants through roots. Granular formulations are sometimes applied by aircraft or helicopter to minimize drift or penetration into dense vegetation. After application, the granules may slowly release the active ingredient. Some granules require soil moisture to release the active ingredient. Granular formulations are also used to control mosquito larvae and other aquatic pests. The pellets are used in pest control operations in agriculture, building treatment, ornamental and lawn plants, aquatic environments, right-of-way, and public health (for stinging insects).

In some cases, the composition is in the form of pellets. Most pellet formulations are very similar to granular formulations; these terms are used interchangeably. However, in a pelletized composition, all particles have the same mass and shape. The homogeneity of the particles facilitates application with precision production equipment.

Powders

In some cases, the composition is formulated as a powder. In some cases, the composition is formulated as a wettable powder. Wettable powders are dry, finely divided formulations that appear as pulverized preparations. They usually need to be mixed with water for spray application. However, few products can be used as a pulverulent preparation or as a wettable powder - the choice is up to the consumer. Wettable powders have from about 1 to about 95 weight percent active ingredient; in some cases more than about 50 percent. The particles do not dissolve in water. They settle quickly unless constantly agitated to keep them in suspension. They can be used for most pest problems and in most types of spray equipment where agitation is likely to occur. Wettable powders have excellent residual activity. Due to their physical properties, most of the composition remains on the surface of treated porous materials such as concrete, plaster and untreated wood. In such cases, only water penetrates the material.

In some cases, the composition is formulated as a soluble powder. Soluble powder formulations appear as wettable powders. At the same time, when mixed with water, soluble powders dissolve easily and form a true solution. After their thorough mixing, additional shaking is not required. The amount of active ingredient in soluble powders ranges from about 15 to about 95 percent by weight; in some cases more than about 50 percent. Soluble powders have all the advantages of wettable powders and none of the disadvantages except for the danger of inhalation during mixing.

In some cases, the composition is formulated as water-dispersible granules. Water-dispersible granules, also known as dry flow formulations, are similar to wettable powders except that they are pulverulent and are formulated into small, easily defined granules. Water-dispersible granules must be mixed with water before use. Once in the water, the granules disintegrate into fine particles, similar to wettable powders. The formulation requires continuous agitation to keep it suspended in water. The percentage of active ingredient is high, often as high as 90 percent by weight. Water-dispersible granules have many of the same advantages and disadvantages of wettable powders, except that they are more easily defined and miscible. Due to the low content of particulate matter, they pose a lower inhalation hazard to the consumer during use.

Bait

In some cases, the composition includes a bait. The bait may be in any suitable form such as a solid, paste, pellet or powder form. The bait may also be carried back by the host to the population of said host (eg, colony or hive). The bait can then act as a food source for other members of the colony, thereby providing an effective modulator to a large number of hosts and potentially the entire host colony.

Baits can be provided in a suitable "house" or "trap". Such houses and traps are commercially available and existing traps can be adapted to include the compositions described herein. The house or trap may be provided, for example, in the form of a box, and may be provided ready-made, or may be made, for example, from folding cardboard. Suitable materials for the house or trap include plastics and cardboard, in particular corrugated cardboard. The interior surfaces of the traps may be lined with a sticky substance to limit the movement of the host when it enters the trap. The house or trap may contain a suitable groove on the inside that can hold the bait in place. A trap differs from a house in that the host cannot easily leave the trap after being hit, while the house acts as a "feeder" that provides the host with a preferred environment in which they can feed and feel safe from predators.

Attractants

In some cases, the composition contains an attractant (eg, a chemoattractant). The attractant may attract an adult host or an immature host (eg a larva) to a location in the vicinity of the composition. Attractants include pheromones, chemicals that are secreted by animals, especially insects, that affect the behavior or development of other individuals of the same species. Other attractants include sugar and protein hydrolysate syrups, yeast fungi, and rotting meat. Attractants can also be combined with the active ingredient and sprayed onto leaves or other objects in the area to be treated.

Various attractants are known to influence host behavior, such as the host's search for food, oviposition or mating sites, or mating partners. Attractants useful in the methods and compositions described herein include, for example, eugenol, phenethyl propionate, ethyldimethylisobutylcyclopropanecarboxylate, propylbenzodioxanecarboxylate, cis-7,8-epoxy-2-methyloctadecane, trans-8,trans-0-dodecadienol, cis -9-tetradecenal (and cis-11-hexadecenal), trans-11-tetradecenal, cis-11-hexadecenal, (Z)-11,12-hexadecadienal, cis-7-dodecenyl acetate, cis-8-dodecenyl acetate, cis-9 -dodecenyl acetate, cis-9-tetradecenyl acetate, cis-11-tetradecenyl acetate, trans-11-tetradecenyl acetate (and cis-11), cis-9,trans-11-tetradecadienyl acetate (and cis-9,trans-12), cis-9 ,trans-12-tetradecadienyl acetate, cis-7,cis-11-hexadecadienyl acetate (and cis-7,trans-11), cis-3,cis-13-octadecadienyl acetate, trans-3,cis-13-octadecadienyl acetate, anethole, and isoamyl salicylate .

Means other than chemoattractants can also be used to attract insects, including lighting devices with different wavelengths or colors of light.

Nanocapsules/Microencapsulation/Liposomes

In some instances, the composition is provided as a microencapsulated formulation. The microencapsulated formulations are mixed with water and sprayed in the same manner as other sprayable formulations. After spraying, the plastic coatings break down and slowly release the active ingredient.

carriers

Any of the compositions described herein may be formulated with the inclusion of the modulating agent described herein and an inert carrier. Such a carrier may be a solid carrier, a liquid carrier, a gel-like carrier and/or a gaseous carrier. In certain embodiments, the carrier may be a seed coater. A seed coater is any non-naturally occurring compound that adheres, in whole or in part, to the surface of a seed. The composition may further contain an excipient or surfactant. The composition may also contain one or more modulators to broaden the spectrum of action.

The solid carrier used for the formulation includes fine clay powder or granules (such as kaolin clay, diatomaceous earth, bentonite clay, Fubasami clay, acid clay, etc.), synthetic hydrated silica, talc, ceramic materials, others inorganic minerals (for example, sericite, quartz, sulfur, activated carbon, calcium carbonate, hydrated silica, etc.), a substance that can be sublimated and is in solid form at room temperature (for example, 2,4,6- triisopropyl-1,3,5-trioxane, naphthalene, p-dichlorobenzene, camphor, adamantane, etc.); wool; silk; cotton; hemp; pulp; synthetic resins (for example, polyethylene resins such as low density polyethylene, unmodified low density polyethylene and high density polyethylene; ethylene-vinyl ester copolymers such as ethylene-vinyl acetate copolymers; ethylene-methacrylic acid ester copolymers such as ethylene and methyl methacrylate and ethylene-ethyl methacrylate copolymers; ethylene-acrylic acid ester copolymers such as ethylene-methyl acrylate copolymers and ethylene-ethyl acrylate copolymers; ethylene-vinylcarboxylic acid copolymers such as ethylene-acrylic acid copolymers; ethylene-tetracyclododecene copolymers; polypropylene resins such as as propylene homopolymers and propylene-ethylene copolymers; poly-4-methylpentene-1, polybutene-1, polybutadiene, polystyrene; acrylonitrile-styrene copolymer resins; styrene elastomers such as acrylonitrile-butadiene-styrene copolymer resins, block copolymers styrene and conjugated dienes and hydrides of yuloc copolymers of styrene and conjugated dienes; fluororubbers; acrylic resins such as poly(methyl methacrylate); polyamide resins such as nylon 6 and nylon 66; polyester resins such as polyethylene terephthalate, polyethylene naphthalate, polybutylene terephthalate and polycyclohexylenedimethylene terephthalate, polycarbonates, polyacetals, polyacrylsulfones, polyarylates, hydroxybenzoate polyesters, polyetherimides, polyester carbonates, polyphenylene ether resins, polyvinyl chloride, polyvinylidene chloride, polyurethane and porous resins such as polyurethane foam or polyurethane foam, polyurethane foam and polyethylene foam etc.), glasses, metals, ceramic materials, fibrous materials, fabrics, knitted fabrics, sheets, paper, yarn, foam, porous substances and multifilaments.

The liquid carrier may include, for example, aromatic or aliphatic hydrocarbons (e.g., xylene, toluene, alkylnaphthalene, phenylxylylethane, kerosene, gas oil, hexane, cyclohexane, etc.), halogenated hydrocarbons (e.g., chlorobenzene, dichloromethane, dichloroethane, trichloroethane etc.), alcohols (such as methanol, ethanol, isopropyl alcohol, butanol, hexanol, benzyl alcohol, ethylene glycol, etc.), ethers (such as diethyl ether, ethylene glycol dimethyl ether, diethylene glycol monomethyl ether, monoethyl ether diethylene glycol, propylene glycol monomethyl ether, tetrahydrofuran, dioxane, etc.), esters (e.g. ethyl acetate, butyl acetate, etc.), ketones (e.g., acetone, methyl ethyl ketone, methyl isobutyl ketone, cyclohexanone, etc.), nitriles (e.g. acetonitrile, isobutyronitrile, etc.), sulfoxides (e.g., dimethyl sulfoxide, etc.), amides (e.g., N,N-dimethylformamide, N,N-dimethylacetamide, cyclic imides (e.g., N-methylpyrrolidone) , alkylidenka carbonates (such as propylene carbonate, etc.), vegetable oil (such as soybean oil, cottonseed oil, etc.), vegetable essential oils (such as orange oil, hyssop oil, lemon oil, etc.), or water.

The carrier gas may include, for example, butane gas, polytetrafluoroethylene gas, liquefied petroleum gas (LPG), dimethyl ether, and carbon dioxide gas.

Excipients

In some cases, the composition provided herein may contain an excipient. Excipients are chemicals that do not have activity. Auxiliaries are pre-blended in the formulation or added to the spray tank to improve mixing or application or to enhance performance. They are widely used in products intended for leaf applications. Excipients can be used to tailor the formulation to suit specific needs and adjust for local conditions. Auxiliaries may be designed to perform specific functions, including wetting, spreading, adhesion, reducing evaporation, reducing volatilization, buffering, emulsifying, dispersing, reducing spray drift, and reducing foaming. No single excipient can perform all of these functions on its own, however, compatible excipients can often be combined to perform multiple functions simultaneously.

Among non-limiting examples of excipients included in the composition are binders, dispersants and stabilizers, in particular, for example, casein, gelatin, polysaccharides (for example, starch, gum arabic, cellulose derivatives, alginic acid, etc.), derivatives lignin, bentonite, sugars, synthetic water-soluble polymers (e.g. polyvinyl alcohol, polyvinylpyrrolidone, polyacrylic acid, etc.), PAP (acid isopropyl phosphate), BHT (2,6-di-tert-butyl-4-methylphenol), BHA (mixture of 2-tert-butyl-4-methoxyphenol and 3-tert-butyl-4-methoxyphenol), vegetable oils, mineral oils, fatty acids and fatty acid esters.

Surfactants

In some cases, the composition provided herein contains a surfactant. Surfactants, also called wetting agents and spreaders, physically change the surface tension of a spray droplet. In order for the composition to properly perform its function, the drop of spray must be able to wet the leaves and be distributed evenly over the leaf. Surfactants increase the area covered by the formulation, thereby increasing the effect of the chemical on the pest. Surfactants are particularly important when applying the formulation to waxy or hairy leaves. Without proper wetting and distribution, spray drops often run off or fail to coat leaf surfaces properly. At the same time, too much surfactant can cause excessive surface runoff and reduce efficiency.

Surfactants are classified based on the way they are ionized or broken down into electrically charged atoms or molecules called ions. A surfactant with a negative charge is anionic. A surfactant with a positive charge is cationic, and a surfactant without an electrical charge is non-ionic. The activity of a formulation in the presence of a nonionic surfactant may differ significantly from that in the presence of a cationic or anionic surfactant. Choosing the wrong surfactant can reduce the effectiveness of the pesticide product and harm the target plant. Anionic surfactants are most effective when used with contact pesticides (pesticides that control the pest by direct contact rather than systemic absorption). Cationic surfactants should never be used as stand-alone surfactants as they are usually phytotoxic.

Nonionic surfactants, often used with systemic pesticides, make it easier for pesticide sprays to penetrate plant cuticles. Nonionic surfactants are compatible with most pesticides, and for most EPA registered pesticides that require a surfactant, the nonionic type is recommended. Adjuvants include, but are not limited to, adhesives, dry diluents, plant penetration agents, compatibility factors, buffers or pH modifiers, drift control additives, antifoams, and thickeners.

Among non-limiting examples of surfactants included in the compositions described herein are salts of alkyl sulfate esters, alkyl sulfonates, aclylaryl sulfonates, alkyl aryl ethers and their polyoxyethylene products, polyethylene glycol ethers, polyhydric alcohol esters, and sugar alcohol derivatives.

Combinations

In formulations and in use forms derived from these formulations, the modulating agent may be present in admixture with other active compounds such as pesticides (e.g., insecticides, sterilizers, acaricides, nematicides, molluscicides, or fungicides; see, for example, pesticides listed below). in table 12), attractants, growth regulators or herbicides. As used herein, the term “pesticidal agent” refers to any substance or mixture of substances intended to prevent, interfere with, repel, or lessen the effects of any pest. A pesticide may be a chemical or biological agent used against pests, including insects, pathogens, weeds, and microbes, that compete with humans for food, destroy property, spread disease, or are a nuisance. The term "pesticidal agent" may further encompass other biologically active molecules such as antibiotics, antiviral pesticides, antifungal agents, anthelmintic agents, nutrients, pollen, sucrose, and/or agents that stop or slow down the movement of insects.

In cases where the modulating agent is applied to plants, admixture with other known compounds such as herbicides, fertilizers, growth regulators, safeners, chemical signal agents or other plant improvers is also possible.

Delivery

The host described herein may be exposed to any of the compositions described herein in any suitable manner that delivers or administers the composition to the insect. The modulator can be delivered alone or in combination with other active or inactive substances and can be applied, for example, by spraying, microinjection, through plants, by watering, dipping, in the form of concentrated liquids, gels, solutions, suspensions, sprays, powders, pellets, briquettes, tiles, etc. formulated to deliver an effective concentration of the modulating agent. The amounts and locations for application of the compositions described herein are generally determined by the habitat conditions of the host, the stage of the life cycle at which host microorganisms can be targeted by the modulating agent, the site at which the application is to be performed, and the physical and functional characteristics of the modulator. The modulators described herein may be administered to the insect by oral ingestion, but may also be administered by agents that penetrate the cuticle or enter the respiratory system of the insect.

In some cases, the insect may simply be "soaked" or "sprayed" with a solution containing a modulating agent. Alternatively, the modulating agent may be associated with a food component (eg, edible) for the insect to facilitate delivery and/or to increase absorption of the modulating agent by the insect. Methods for oral administration include, for example, direct mixing of the modulator with insect food, spraying the modulator into the insect's habitat or field, and design-based approaches in which the species to be used as food is engineered to express the modulator. means, after which it is fed to an insect that is subject to negative influence. In some cases, for example, the modulating agent composition may be included in or topped off the insect's diet. For example, the modulating agent composition can be sprayed onto a crop field in which the insect lives.

In some cases, the composition is sprayed directly onto the plant, e.g. crops, e.g. by backpack spraying, aerial spraying, crop spraying/pollination, etc. In cases where the modulating agent is delivered to the plant, the plant receiving the modulating agent , can be at any stage of plant growth. For example, the formulated modulators can be applied as a seed coat or root treatment in the early stages of plant growth, or as a full plant treatment in the later stages of the crop cycle. In some instances, the modulating agent may be applied to the plant as a topical agent such that the host insect ingests or otherwise comes into contact with the plant upon interaction with the plant.

In addition, the modulating agent can be applied (for example, to the soil in which the plant grows, or to the water used to water the plant) as a systemic agent that is absorbed and distributed throughout the tissues (for example, stems or leaves) of the plant - the host or host animal in such a way that the insect feeding on it will receive an effective dose of the modulating agent. In some instances, food plants or organisms can be genetically engineered to express the modulating agent such that a host feeding on the food plant or organism ingests the modulating agent.

Sustained or sustained release can also be achieved by coating the modulating agent or composition containing the modulating agent(s) with a soluble or biodegradable coating layer such as gelatin, which coating dissolves or breaks down in the application environment, which then makes the modulating agent available, or by dispersing the agent in a soluble or degradable matrix. Such continuous release and/or distribution means that the devices can preferably be used to sustainably maintain an effective concentration of one or more of the modulators described herein in a particular host environment.

The modulating agent may also be included in the environment in which the insect grows, lives, breeds, feeds, or infects. For example, the modulating agent may be included in a food container, feeder, protective wrap or hive. In some applications, the modulating agent may be associated with a solid support for use in powder form or in a "trap" or "feeder". By way of example, in applications in which the composition is to be used as a trap or bait for a particular insect host, the compositions may also be bound to a solid support or encapsulated in a slow release material. For example, the compositions described herein can be administered by delivering the composition to at least one habitat where the insect grows, lives, breeds, or feeds.

Screening

Included herein are screening assays for identifying a modulator where the modulator is effective in altering the host microbiota and thereby enhancing host fitness (eg, insect fitness). For example, a screening assay can be used to identify one or more modulators that target specific microorganisms and/or specific hosts. Additionally, screening assays can be used to identify one or more microorganisms with improved functionality. For example, a screening assay may be effective in isolating one or more microorganisms with an improved ability to metabolize (e.g., degrade) a pesticide (e.g., an insecticide, e.g., neonicotinoid) or allelochemicals in a plant (e.g., caffeine, soy cystatin N, monoterpenes, diterpenic acids). or phenolic compounds). Delivery and colonization of the isolated microorganism in the host can increase the host's resistance to pesticides or allelochemicals in the plant, thereby increasing the host's fitness. The methods may also be useful for isolating microorganisms with an improved ability to colonize any of the hosts described herein.

For example, to screen for microorganisms that increase host resistance to a pesticide, a stock culture that includes microorganisms (e.g., bacteria) and high concentrations of a pesticide (e.g., the pesticide listed in Table 12 or a pesticide known in the art, e.g. , neonicotinoid). In some cases, the pesticide may be provided in the form of a sample of the pesticide-enriched medium (eg, a soil sample). Alternatively, the pesticide (eg, the pesticide listed in Table 12) may be provided in pure form or in combination with other carriers. Additionally, one or more microbial isolates may be inoculated directly into the medium (eg, from a laboratory strain) or may be a sample medium containing one or more microorganism species. The growth medium may be liquid or solid. In some cases, the pesticide of interest is the only source of carbon or nitrogen for microorganisms in the environment. The culture can be subcultured (eg, inoculated into fresh medium with high pesticide levels) any number of times to enrich and/or isolate strains of microorganisms (eg, bacterial strains) capable of metabolizing the pesticide. The parent culture or subcultures can be evaluated using any of the methods known in the art to investigate changes (eg, reduction) in pesticide levels in a sample (eg, using HPLC). Isolates that reduce the concentration of a pesticide (eg, the pesticide listed in Table 12 or a pesticide known in the art, eg, neonicotinoid) can be isolated for use as a modulator in any of the methods or compositions described herein.

The methods can be used to further select for the microorganisms described herein, including those isolated from screening assays, with improved ability to colonize and survive in a host (eg, insect). For example, the host may be inoculated with a bacterial isolate (eg, an isolate with the ability to degrade a pesticide). The host can then be examined at regular intervals to check for the presence of a bacterial isolate (eg, by culture or by 16s RNA from intestines isolated from the host). Bacterial isolates that survive within the host (eg, in the insect midgut) can be isolated for use as a modulator in any of the methods or compositions described herein.

Table 12. Pesticides Aclonifen Fenchlorazole-ethyl Pendimethalin Acetamiprid Fenotiocarb Penflufen Alanicarb Fenitrothion Penflufen Amidosulfuron fenpropidine Pentachlorobenzene Aminocyclopyrachlor Fluazolate Penthiopyrad Amisulbrom Flufenoxuron Penthiopyrad Anthraquinone flumetralin Pirimiphos-methyl Asulam, sodium salt Fluxapiroxade Pralletrin Benfuracarb Fuberidazole Profenofos Bensulide ammonium glufosinate Proquinazid Beta HCH; beta BCH Glyphosate Prothiophos Bioresmetrin Group: Borax, borate salts (see. Pyraclophos Blasticidin-S Group: Paraffin oils, mineral Pyrazachlor Bura; disodium tetraborate Gulfenprox Pyrazophos Boric acid Imiprotrin pyridaben Bromoxynil heptanoate Imidacloprid Pyridalyl Bromoxynil octanoate Ipconazole Pyridifention Carbosulfan isopyrazam Pyrifenox Chlorantraniliprole isopyrazam Quinmerak Chlordimeform Lenacil Rotenone Chlorfluazuron magnesium phosphide Sedaksan Chlorpropham Metaflumizone Sedaksan Climbazole Metazachlor Silafluofen Clopyralid Metazachlor Sintofen Copper(II) hydroxide Metobromuron Spinetors Cyflufenamid Methoxuron Sulfoxaflor Cyhalothrin Metsulfuron-methyl Temephos Cyhalothrin, gamma Milbemectin thiacloprid Decahydrate Naled Thiamethoxam Diafenthiuron Napropamide Tolfenpirad Dimefuron Nicosulfuron Tralomethrin Dimoxistrobin Nitenpiram Tributyltin compounds dinotefuran Nitrobenzene Tridifan Diquat dichloride o-phenylphenol Triflumizole Dithianon Oils Validamycin E-phosphamidon Oksadiargyl Zinc Phosphide EPTC Oxycarboxyne Etaboxam Paraffin oil Etirimol Penconazole

EXAMPLES

The following is an example of the methods of the present invention. It should be understood that various other embodiments may be practiced given the general description above.

Example 1. Formation of a library of natural microorganisms

This example demonstrates the isolation of soil bacteria that naturally produce an amino acid, namely methionine.

The medium used for isolation of microorganisms is starch-casein-nitrate agar (starch, 10.0 g; casein, 0.003 g; KNO ₃ , 0.02 g; NaCl, 0.02 g; MgSO ₄ , 0.5 mg ; CaCO ₃ , 0.2 mg; FeSO ₄ , 0.1 mg; agar, 12.0 g; H ₂ O, 1 L; pH 7.0) (Kuster and Williams, 1964). Each medium sample representing a soil sample (1.0 g) was suspended in 9 ml of sterile water and 1 ml of the suspension was serially diluted ten times in sterile distilled water. One milliliter of a 10 ^-5 dilution was inoculated into agar medium and incubated for 7 days at 30°C. At the end of this period, the plates were checked for growth. White individual and dense colonies were collected and grown on new starch casein nitrate agar plates to form a library of isolates. After 7 days of growth at 30°C, the cups were stored at 4°C.

Example 2 Screening for Isolates That Produce Methionine

This example demonstrates a screening assay of the isolates from Example 1 that naturally produce the amino acid, methionine.

Screening for methionine production

Modified basal medium (K ₂ HPO ₄ 0.3 g; KH ₂ PO ₄ 0.7 g; Na ₂ CO ₃ 1.0 g; CaCl ₂ 5.0 mg; MgSO ₄ 0.3 g; FeSO ₄ , 1.0 mg; H ₂ O, 1 L) containing sucrose (20.0 g) and NH ₄ Cl (10.0 g) is used for fermentation (Chay, BP, Galvez, FCF, and Padolina, WGPULBP (1992) Production of Methionine by Batch Fermentation from Various Carbohydrates, ASEAN Food Journal (Malaysia). The pH of the medium was 7.2.

Culture Conditions Two loops of a 7 day old culture of the isolate from Example 1 were inoculated into a 250 ml Erlenmeyer flask containing 30 ml of fermentation medium. Methionine production was analyzed after the flask was incubated for 5 days on a rotary shaker (160 rpm) at 30°C. Double flasks were prepared and uninoculated flasks served as controls in all experiments.

The presence of methionine in the broth cultures of the isolates was examined by paper chromatography according to a modified method of Khanna and Nag (Khanna et al., "Production of amino acids in vitro tissue culture", Indian Journal of Experimental Biology (1973)). The broth culture was centrifuged at 5000 xg for 20 minutes. and 2 µl of the supernatant was applied 1.5 cm above one edge of 18 cm x 22 cm Whatman No. 1 filter paper. in a solvent mixture containing n-butanol, acetic acid and water (4:1:1) for 18 hours. The chromatogram was dried in air at room temperature, sprayed with a 0.15% solution of ninhydrin in butanol and dried again before heating at 60° C for 5 min. in the oven. The value of the color index of the ninhydrin-positive spot (bluish-purple) of the supernatant, which corresponds to the value of the standard solution of methionine, indicated the presence of methionine in the broth culture. The concentration of methionine produced in the broth culture of the isolate was evaluated as follows. The ninhydrin-positive supernatant spot of the isolate on the chromatogram was eluted in 10% ethanol and the spectrophotometric reading of the eluate was recorded at 520 nm. The concentration of methionine in the supernatant was determined from the standard curve. A graph of optical density values in relation to various concentrations (0.1-0.9 mg/ml) of methionine solution served as a standard methionine curve.

Isolates that produce methionine were stored on fresh agar plates and a stock solution was formed by suspending two micro-inoculation loops in an aliquot of 50% glycerol solution.

Example 3 Administration of Methionine Producing Isolates to Increase Amino Acid Content in Crickets

This example demonstrates the ability of methionine-producing bacteria to increase their nutrient content when treated with crickets.

The global demand for meat is increasing and the supply of animal feed is placing increasing pressure on land and water resources. Insects can provide the large amount of protein that animals need with much less environmental impact; many species of insects can feed on compost, such as Grant's larvae, or other types of organic waste, such as leftover food, by-products of processing, and grain discarded by breweries. Insects gain weight at an amazing rate, in part because, as cold-blooded animals, they don't expend energy regulating their body temperature. Crickets, such as Acheta domesticus , need only 1.7 kilograms of food to put on a kilogram of body weight; a typical US chicken consumes 2.5 kilograms, pigs consume 5 kilograms, and cattle 10 kilograms. Another advantage: most insects can be eaten whole. Only about half of a chicken or pig is edible; for cows, the proportion is even less. As a result, growing a kilogram of insect protein produces less CO ₂ than raising pigs or cattle, and requires only one tenth of the land resources.

Insect foods can replace 25% to 100% of soy products or fishmeal in animal diets without adverse effects, but most insect foods lack the amino acids methionine and lysine. Synthetic production of methionine requires hazardous chemicals and their use is prohibited in organic farming. By introducing methionine-producing bacteria into the cricket microbiota, crickets are expected to naturally increase their nutrient content.

Therapy regimen The isolated bacteria, identified as methionine-producing bacteria from Example 2, were formulated with a solution of 10 ⁷ cells/ml, mixed with a nutrient substrate, eg poultry starter feed and rice bran (Poultry Feed-PF) for crickets.

Experiment scheme

The cricket experimental setups are modified shipping boxes with a removable bottom and lid that have the base area of standard pallets used for international shipping (1.2 m (L) x 1.0 m (W) x 0, 61 m (H)). The interior of each enclosure is lined with a 4mm clear plastic liner and covered with a 122cm x 137cm nylon mosquito net that acts as a physical barrier to entry or exit. To prevent cannibalism and stress-related mortality, 96 30cm x 30cm egg cartons were placed at the edge of each shipping carton. This provides approximately 172,800 cm ^{2 of} visible surface area. Water access was provided by 2-quart poultry water dispensers with cotton and gravel added to the distribution tank to prevent newly hatched nymphs from drowning. The sides of the water dispensers were sprinkled with sand to provide a foothold for the crickets to crawl vertically. Spray tips with check valves to prevent dripping were attached to the top of each housing. To maintain acceptable humidity and provide an alternative source of water with the ability to spray for a large population of crickets, such tips provided water bursts aimed at the center of the housing at automatically set intervals. Temperature (T) and relative humidity (RH) in the greenhouse were maintained at 29.0 ± 2.1 standard deviation (SD) °C and 67.2 ± 14.7 SD%, respectively, during the experiment. Light was provided 24 hours a day.

An egg substrate from Timberline Fisheries (http://timberlinefresh.com), consisting of approximately 50,000 Acheta domesticus eggs with a hatching rate of 70%, was placed in each of the boxes. The humidity of the egg substrate was maintained at 80-90% until hatching. When hatching was observed, the substrate was irrigated with water twice a day until the final emergence of the nymphs. Population growth was monitored every 3-4 days by counting and weighing a random sample of 70 individuals from each experimental setup.

14 days after hatching and until collection or final death, Acheta domesticus populations were administered the following. 2 feeds for ad libitum treatment: 1) 5:1 ratio of non-medicated poultry feed to rice bran (Poultry Feed-PF) as control; 2) 5:1 ratio of non-medicated poultry feed and rice bran (Poultry Feed-PF) sprayed with 100 ml of a 10 ⁹ cell/ml solution of the isolated bacteria described in Example 2 diluted in the growth medium described herein.

Once a week for five weeks of cultivation, insects were collected. The insects were stored for half an hour in a freezer at -50°C. Next, the frozen insects were immersed in liquid nitrogen and successively ground using a mixer for 15 minutes (Braun Multiquick 5, 600 W, Kronberg, Germany). The amino acid composition of the lyophilized insect powder was analyzed using ion exchange chromatography following the international standard ISO 13903:2005 and the method of Yi, L. et al. (2013).

Crickets fed on a diet containing the methionine-producing microorganisms identified in Example 2 were expected to have higher methionine content than crickets fed on a control diet.

Example 4 Introduction of Methionine Producing Bacterial Strains Drosophila melanogaster grown with methionine deficient food to increase their body weight, development rate and survival

This example demonstrates the ability of methionine-producing bacteria, when treated with Drosophila melanogaster , grown on a diet deficient in methionine, to increase body weight, growth rate, and survival. This experimental design is also applicable to nutrient enhancement of other insects, such as crickets, which can be used to produce methionine-rich animal feed.

Experiment scheme

Bacterial strains isolated in example 2 that produce methionine, as well as strains that do not produce methionine, were grown in nutrient broth at 30°C.

Chemically defined (CD) fly food was prepared as described in Nature, Vol. 11, no. 1, 100-105, 2014. Received CD food, which lacks methionine, and it was called CD-M. Fly food formulations were used for all experiments described in this example.

Growth rate and body weight analyzes

On the first day, 10 ⁹ methionine-producing bacteria described in Example 1 or bacteria that do not produce methionine (control) were resuspended in 100 μl of phosphate-buffered saline and added to CD-M fly food. Two such cohorts were left to dry for 24 hours at 25°C.

On the second day, the embryos obtained as a result of fertilization were collected from flies, treated with 2% hypochlorite solution for 5 min. and then washed with sterile water to remove any extracellular microorganisms from the embryos. 10 µl of a suspension of embryos in water (1:3 embryos:aqueous suspension) was added to both the bacteria-inoculated and control samples. The fly-fed cohorts with embryos were maintained at 25° C. with a cycle of 12 hours light and 12 hours dark until the end of the experiment.

The time of puparia formation and the number of formed pupae were measured in each cohort. The time of emergence of adults and the rate of appearance were measured in each sample. Starting from the moment the first adult emerged from the pupa, the number of adult flies that emerged was counted every 12 hours and the rate of emergence was calculated.

Based on body weight analysis, ten larvae from both cohorts were selected and their masses, areas, and total protein content were measured.

Embryos fed CD-M fly chow inoculated with the methionine-producing bacteria identified in Example 2 were expected to develop faster and have a higher protein content than embryos fed CD-M fly chow with bacteria not producing methionine.

Survival analysis

12 days prior to the first day, sterile embryos were formed as previously described and grown using sterile CD fly chow. Sterile adults began to emerge from their pupae 11 days after embryo collection when grown at 25° C. with a cycle of 12 hours light and 12 hours dark.

On the first day 10 ⁹ methionine producing bacteria or bacteria that do not produce methionine (control) were resuspended in 100 μl of phosphate buffered saline and added to CD-M fly food. Two such cohorts were left to dry for 24 hours at 25°C.

10 newly emerged sterile adult males and females were introduced into CD-M fly chow with methionine producing bacteria or control on the second day of the experiment. The fly food with flies was maintained at 25° C. with a cycle of 12 hours light and 12 hours dark until the end of the experiment. The number of surviving male and female flies was counted every day until all flies had died. A survival assay was performed to evaluate the favorable fitness effects of methionine-producing bacteria on the survival of the flies.

Flies reared with CD-M fly chow inoculated with the methionine producing bacteria identified in Example 2 were expected to survive longer than controls.

Example 5 Isolation of Microorganisms that Degrade Fenitrothion, an Organophosphate Insecticide

This example demonstrates the preparation of a library of microorganisms capable of degrading fenitrothion, an organophosphate insecticide.

Experiment scheme

Soil samples were obtained from various sites where fenitrothion had previously been used for insect control. Fenitrothion degrading bacteria were isolated from soil samples as described in (Microbes Environ. Vol. 21, No. 1, 58-64, 2006). Briefly, 1 g of a soil sample was thoroughly mixed with 9 ml of sterile distilled water. The soil particles were then centrifuged at 1000 rcf for 5 min. and 100 μl of the supernatant was then plated on fenitrothion with a medium of mineral salts (0.4 g/l yeast extract, 0.4 g/l fenitrothion, 15 g/l microbiological agar). The plates were cloudy upon receipt as fenitrothion is an emulsion.

Colonies that form clear patches around them and are capable of degrading or metabolizing fenitrothion were isolated and re-grown on LB agar, nutrient agar, glucose yeast agar, TSA agar, BHI agar or MRS agar. Once unique bacterial colonies were identified, their genome was extracted using a genomic DNA extraction kit (Qiagen, DNeasy blood and tissue collection kit) following the manufacturer's protocol.

The 16S rRNA gene was amplified using universal bacterial primers 27F (5'AGAGTTTGATCMTGGCTCAG-3'; SEQ ID NO: 227) and 1492R (5'-TACCTTGTTACGACTT-3'; SEQ ID NO: 228), and using PCR conditions at 94° C for 2 min., 30 cycles at 94°C for 1 min., at 56°C for 1 min. and at 72°C for 2 minutes, and a final elongation at 72°C for 5 minutes. Gel electrophoresis was used to confirm that the amplicons were of the correct size (~1500 bp) and that the products were removed from the gel and purified using a gel extraction kit (Qiagen, QIAquick) following the manufacturer's protocol. Amplicons of the correct size were Sanger-sequenced and BLAST was used to compare the sequence with different archives of 16s rRNA genetic sequencing data for bacterial identification.

To determine if isolated bacteria cleave fenitrothion, ~10 ⁷ bacterial cells were incubated in 1 ml of 20 mM sodium potassium phosphate buffer (pH 7) with 1 mM fenitrothion, as described in PNAS, Vol. 109, no. 22, 8618-8622, 2012. After 30 min. incubation at 30°C the reaction was stopped by adding an equal volume of methanol. Then fenitrothion and its metabolite, 3-methyl-4-nitrophenol, were analyzed by HPLC. The retention times and peak areas of the HPLC profiles were compared to known standards.

Isolates of unique bacteria that are able to degrade fenitrothion were then stored as frozen glycerol at -80°C.

Example 6: Improving resilience Drosophila melanogaster to fenitrothion through the introduction of bacteria that break down fenitrothion

This example demonstrates the possibility of obtaining a model insect, Drosophila melanogaster , that is resistant to one or more of the negative effects of insecticides in their diet, more specifically fenitrothion, resulting in a more resistant insect. A further approach is to substitute other insects, such as crickets or other insects disclosed herein, for example, insect sources suitable for protein-rich pet food. Many insecticides, including fenitrothion, have been shown to be toxic to crickets.

Experiment scheme

Therapy Schedule The bacterial isolates selected in Example 5 were formulated at 10 ⁹ organisms per 100 µl of fly food medium with and without fenitrothion.

The medium used to feed the flies was a medium containing cornmeal, molasses and yeast (11 g/l yeast, 54 g/l yellow cornmeal, 5 g/l agar, 66 ml/l molasses and 4.8 ml/l l propionic acid). For experimental procedures, fenitrothion at 0, 10, and 100 p.p.m. or phosphate buffered saline as negative controls were infused into a sterile fly food medium.

Development rate analysis

On the first day, 109 ^fenitrothion -producing bacteria described in Example 5 were suspended in 100 µl of phosphate buffered saline or equal volumes of saline (negative controls) were added to sterile fly food media with or without fenitrothion. The samples were left to dry during the day at 25°C, as described in Appl. Environ. microbiol . Vol. 82, no. 20, 6204-6213, 2016.

On the second day, the embryos obtained as a result of fertilization were collected from flies, treated with 2% hypochlorite solution for 5 min. and then washed with sterile water to remove any extracellular microorganisms from the embryos. 10 μl of a suspension of embryos in water (1:3 embryos:aqueous suspension) was added to the fly chow inoculated with bacteria or a negative control chow with or without fenitrothion. The fly-fed cohorts with embryos were maintained at 25° C. with a cycle of 12 hours light and 12 hours dark until the end of the experiment.

The time of puparia formation and the number of formed pupae were measured in each cohort. The time of emergence of adults and the rate of appearance were measured in each sample. Starting from the moment the first adult emerged from the pupa, the number of adult flies that emerged was counted every 12 hours and the rate of emergence was calculated.

It was expected that embryos fed fly chow infused with fenitrothion inoculated with fenitrothion degrading bacteria would develop faster than embryos fed fly chow infused with fenitrothion without fenitrothion degrading bacteria.

Survival Analysis

Approximately 12 days prior to the first day, sterile embryos were formed as previously described and grown using sterile fly chow. Adults began to emerge from their pupae 11 days after harvested embryos were grown on sterile fly chow without fenitrothion at 25° C. with a cycle of 12 hours light and 12 hours dark.

On the first day, 10 ⁹ fenitrothion degrading bacteria in phosphate buffered saline were added to a sterile fly food medium as described in the previous example.

On the second day, 10 newly emerged sterile adult males and females were introduced into fly chow inoculated with bacteria or into a chow which was a negative control with or without fenitrothion. The fly food with flies was maintained at 25° C. with a cycle of 12 hours light and 12 hours dark until the end of the experiment. The number of surviving male and female flies was counted every day until all flies had died. A survival assay was performed to assess the favorable fitness effects of fenitrothion-degrading bacteria on fly survival.

It was expected that flies reared with fly chow infused with fenitrothion inoculated with fenitrothion-degrading bacteria would survive longer than flies reared with fly chow infused with fenitrothion without the degrading bacteria. fenitrothion.

Example 7 Elimination of Entomopathogenic Bacteria in Drosophila melanogaster using naturally occurring phages

This example demonstrates the ability of naturally occurring phages to eliminate bacterial pathogens (such as Serratia marcescens , Erwinia carotovora , and Pseudomonas entomophila ) in the insect Drosophila melanogaster . This procedure can be used as a surrogate assay to eliminate bacteria from other insect species such as bees.

Experiment scheme

Therapy scheme A collection from a phage library containing the following phage families was used: Myoviridae, Siphoviridae, Podoviridae, Lipothrixviridae, Rudiviridae, Ampullaviridae, Bicaudaviridae, Clavaviridae, Corticoviridae, Cystoviridae, Fuselloviridae, Gluboloviridae, Guttaviridae, Inoviridae, Leviviridae, Microviridae, Plasmaviridae, Tectiviridae.

Several media samples (soil, pond sediments and wastewater) were collected in sterile 1 L flasks over a period of 2 weeks and processed immediately after collection and then stored at 4°C. Solid samples were homogenized in Sterile Luria Broth (LB) or Difco Double Strength Tryptic Soy Broth (TSB) supplemented with 2 mM CaCl2 to a final volume of 100 ml. pH values and phosphate levels were measured using phosphate strips. For purification, all samples were centrifuged at 3000–6000 g for 10–15 min. at 4°C and filtered through filters with a pore size of 0.2 μm with low protein binding to remove any remaining bacterial cells. The supernatant, which contains the phage library, was then stored at 4° C. in the presence of chloroform in a glass bottle.

A 20-30 ml phage library was diluted to a volume of 30-40 ml with LB broth. A target bacterial strain (e.g. , Serratia marcescens , Erwinia carotovora , and Pseudomonas entomophila ) was added (50-200 µl of overnight culture in LB broth) to enrich the population of phages that target that particular bacterial strain in culture. This culture was incubated overnight at 37° C., shaking at 230 rpm. Bacteria from this enriched culture were removed by centrifugation (3000-6000 g for 15-20 min. at 4°C) and filtered (0.2 or 0.45 µm filter). 2.5 ml of bacteria-free culture was added to 2.5 ml of LB broth and 50-100 µl of target bacteria were added back to the culture for further phage enrichment. The enriched culture was grown overnight as described above. A sample from this enriched culture was centrifuged at 13,000 g for 15 min. at room temperature and 10 µl of the supernatant was placed on a Petri dish with LB-agar together with 100-300 µl of target bacteria and 3 ml of molten 0.7% soft agar. Cups were incubated overnight at 37°C.

Each of the plaques observed on the bacterial lawn was collected and transferred to 500 μl of LB broth. A sample from this initial plaque culture was additionally plated on target bacteria. Plaque cleaning was performed three times for all detected phages in order to isolate one homogeneous phage from a heterogeneous mixture of phages.

Phage lysates with high titers (> 1x 10^10 pfu/ml) were prepared by collecting top layers from plates containing a host bacterial strain showing confluent lysis. After adding 5 ml of buffer, the top soft agar layer was macerated, purified by centrifugation and sterilized by filtration. The resulting lysates were stored at 4°C. High titer phage lysates were further purified by CsCl isopycnal centrifugation as described in Summer et al., J. Bacteriol . 192:179-190, 2010.

DNA was isolated from CsCl-purified phage suspensions as described in Summer, Methods Mol. Biol. 502:27-46, 2009. A single isolated phage was sequenced as part of two pools of phage genomes using pyrosequencing method 454. Briefly, phage genomic DNA was mixed in equimolar amounts to a final concentration of approximately 100 ng/l. The pooled DNA was fragmented, ligated with a multiplex identification tag (MID) specific to each of the pools, and sequenced by pyrosequencing using a full plate reaction on a sequencer (Roche) according to the manufacturer's protocols. Pooled phage DNA was present in two sequencing reactions. The output corresponding to each of the pools was collected separately using software (454 Life Sciences), adjusting the settings so that only reads with one MID per build were included. The identity of individual contigs was determined by PCR using primers generated for the contig sequences and individual phage genomic DNA preparations as a template. Sequence software (Gene Codes Corporation) was used to assemble and edit sequences.

Chromosomal terminal structures of phages were determined experimentally. Sticky ends (cos) for phages were determined by sequencing the ends of the phage genome and sequencing PCR products obtained by amplification with a ligation compound of circularized genomic DNA as described in Summer, Methods Mol. Biol. 502:27-46, 2009. Protein-coding regions originally predicted using gene prediction software (Lukashin et al. Nucleic Acids Res. 26:1107-1115, 1998), purified by manual analysis in Artemis (Rutherford et al ., Bioinformatics 16:944-945, 2000) and analyzed using BLAST (cut-off E-value 0.005) (Camacho et al., BMC Bioinformatics 10:421, 2009). Proteins of particular interest were further analyzed with sequence search software (Hunter et al., Nucleic Acids Res . 40:D306-D312, 2012).

Electron microscopy of CsCl-purified phage (>1 x10^11 PFU/mL) that lysed pathogenic bacterial speciesDrosophila, was performed by diluting the stock phage solution with tryptic soy broth buffer. The phages were applied to 400 mesh carbon-coated fine meshes, stained with 2% (w/v) uranyl acetate, and air dried. The samples were observed in a transmission electron microscope operating at an accelerating voltage of 100 kV. Five virions of each phage were measured to calculate the average values and standard deviations of the capsid and process sizes, if necessary.

Introduction of phages into food

The medium used to feed the flies was a medium containing cornmeal, molasses and yeast (11 g/l yeast, 54 g/l yellow cornmeal, 5 g/l agar, 66 ml/l molasses and 4.8 ml/l l propionic acid). The phage solutions were infused into the fly feed to give final phage concentrations of 0 to 10 ⁸ PFU/mL.

The bacteria S. Marcescens , Erwinia carotovora and Pseudomonas entomphila were grown at 30°C in nutrient broth or LB broth.

Sterile fly embryos were obtained by treating fertilization embryos collected from flies with a 2% hypochlorite solution for 5 minutes. and then washed with sterile water to remove any extracellular microorganisms. Fly larvae with target bacteria were obtained by incorporating 10 ⁹ CFU of bacteria into a sterile fly diet and adding sterile fly embryos to the diet. After 2 days, ten infected larvae of the first instar flies S. marcescens were added to the fly feed in the range (0-10 ⁸ PFU/ml) of phage concentrations. The larvae were allowed to grow in the phage feed for 3 days until they reached the third instar stage. Then the larvae were collected and individually homogenized in nutrient broth or LB broth and placed on nutrient agar or LB agar plates and incubated at 30°C. The number of CFUs of S. marcescens obtained from larvae in fly food with different phage concentrations was recorded. This indicated the number of live bacteria that were present in the flies.

It was expected that the number of live bacteria would be reduced in larvae reared with phage fly chow compared to those with bacteria.

Example 8 Introduction of an Amino Acid Producing Bacteria Strain Drosophila melanogaster through diet to increase their rate of development

This example demonstrates the ability of amino acid producing bacteria to increase their nutrient content when treated with the insect Drosophila melanogaster . This experimental design can be extended to reduce growth time and obtain more biomass from other insects, such as crickets, which can be used to produce protein-rich animal feed.

The global demand for meat is increasing and the supply of animal feed is placing increasing pressure on land and water resources. Insects can provide the large amount of protein that animals and humans need at much lower environmental impact; many species of insects can feed on compost, such as Grant's larvae, or other types of organic waste, such as leftover food, by-products of processing, and grain discarded by breweries. Insects gain weight at an amazing rate, in part because, as cold-blooded animals, they don't expend energy regulating their body temperature. Crickets, such as Acheta domesticus , need only 1.7 kilograms of food to put on a kilogram of body weight; a typical US chicken consumes 2.5 kilograms, pigs consume 5 kilograms, and cattle 10 kilograms. Another advantage: most insects can be eaten whole. Only about half of a chicken or pig is edible; for cows, the proportion is even less. As a result, growing a kilogram of insect protein produces less CO ₂ than raising pigs or cattle, and requires only one tenth of the land resources.

Insect food can replace 25% to 100% of soy products or fishmeal in animal diets without adverse effects. However, most insect foods lack the amino acids methionine and lysine. Synthetic production of methionine requires hazardous chemicals and their use is prohibited in organic farming. In this example, the introduction of methionine-producing bacteria into the microbiota of an insect naturally increases their nutrient content.

Therapy regimen

The isolated bacteria Corynebacterium glutamicum that produce glutamate or methionine were formulated with a solution of 109 colony forming units (CFU) mixed with Drosophila fly nutrient substrate.

Experiment scheme

Strains of Corynebacterium glutamicum that produce glutamate or methionine were grown in nutrient broth at 30°C.

The medium used to feed the flies was a medium containing cornmeal, molasses and yeast (11 g/l yeast, 54 g/l yellow cornmeal, 5 g/l agar, 66 ml/l molasses and 4.8 ml/l l propionic acid). All components of the diet, with the exception of propionic acid, were heated together to 80°C in deionized water with constant stirring for 30 minutes, and they were allowed to cool to 60°C. Propionic acid was then mixed in and 50 ml of the ration was aliquoted into separate vials and left to cool and solidify. Flies were reared at 26°C, light:dark cycle 16:8 hours, with a humidity of about 60%.

For an experimental setup to measure larval growth rate, a fixed diet was used (Piper et al., 2014, Nature Methods ). The established diet eliminated the effects of intergroup variation in the ingredients used for the cornmeal diet. In addition, the established diet allowed the exclusion of individual components in order to study their effects on the development of flies.

Development rate analysis

On the first day, 100 μl of a suspension of Corynebacterium glutamicum in 10 ⁹ colony forming units (CFU) nutrient broth was added to five replicates of the fly food. As controls, bacteria-free nutrient broth was added to five more vials of fly food. Embryos resulting from fertilization were collected from fruit flies, which were treated with 2% hypochlorite solution for five minutes and then washed with sterile water to remove any extracellular microorganisms from the embryos. 10 µl of embryo-in-water suspension (one:three embryos:water suspension) was added to all fly food bottles, both inoculated with bacteria and controls. Feed for flies with embryos was maintained at 26°C, light:dark cycle of 16:8 hours at about 60% humidity until the end of the experiment. The time of emergence of adults and the rate of appearance were measured in each repetition. Starting from the moment the first adult emerged from the pupa, the number of adult flies that emerged was counted every 12 hours and the rate of emergence was calculated.

Analysis of the mass of larvae

To test whether the presence of amino acid-producing bacteria could provide an increase in the body weight of developing larvae when reared on a particular diet, axenic and single-strain-associated larvae were generated. Three different bacteria were used for these assays, Corynebacterium glutamicum, a strain that produces glutamate, Corynebacterium glutamicum, a strain that produces methionine, and E. coli .

First, axenic embryos were formed. Embryos resulting from fertilization were collected from fruit flies over a period of 6 hours on plates containing wort agar with yeast. To eliminate any bacterial contamination, the embryos were treated with 2% hypochlorite solution for five minutes and then washed three times with sterile water. One volume of embryos was then suspended in 3 volumes of water.

The established diet was aliquoted into vials and nine replicates were used for each test condition. Investigated conditions:

1. food without added bacteria;

2. food containing a C. glutamicum strain that produces glutamate (C.glu-Glu);

3. feed containing a strain of C. glutamicum that produces methionine (C.glu-Met);

4. food containing E. coli .

To each of the food vials that were in conditions 2, 3, and 4, 100 µl of overnight stationary phase culture was added.

To each of the nine replicates in each condition, 10 μl of a sterile germ+water suspension was added. The vials were then incubated at 26° C., 60% humidity and a 16:8 light:dark cycle.

After 13 days, 10-15 larvae were selected randomly from each replication and their areas were measured as an indicator of their biomass and weight. The larvae were scraped from the food with a sterile spatula, washed in water to clear the food of their bodies, and each larva was imaged separately for each replicate in each condition. The Image J program was used to identify, describe and measure the area of the larva in each image.

Due to the treatment with bacteria producing the amino acid, the rate of development of the insect increased.

Embryos that developed using a diet seeded with an amino acid-producing bacterial strain reached maturity significantly faster than embryos that were grown using a sterile diet (FIG. 1). Additionally, this effect was slightly stronger in female flies than in male flies (FIGS. 2A and 2B).

An increase in the body weight of the larva due to treatment with bacteria that produce the amino acid.

Larvae reared on a diet supplemented with C. glu-Met averaged the largest body size, followed by larvae reared on a diet supplemented with C. glu-Glu, E. coli and without bacteria (Fig. 3 ). This shows that supplementing the insect diet with bacteria that produce amino acids allows insect biomass to be obtained faster than using the diet without amino acid supplementation.

Together, these data demonstrate that supplementing the insect diet with bacteria that are capable of producing amino acids allows insect biomass to be obtained faster than using the diet without amino acid supplementation. Extending this to other insects such as crickets, supplementing their diet with bacteria that are capable of producing methionine, can provide an increase in their biomass and protein content.

Example 9 Insects Treated with Purified Phage Solution

This example demonstrates the isolation and purification from media samples of phages that have targeted specific insect bacteria. This example also demonstrates the efficacy of isolated phages against target bacteria in in vitro plaque assays, as measured by their oxygen consumption rate and extracellular acidification rate. Finally, this example demonstrates the effectiveness of phages in vivo, as measured by the ability of the phage to target the bacteria of flies by treating them with the isolated phage that counteracts the bacteria. This example demonstrates that a pathogenic bacterium that reduced the fitness of an insect could be eliminated by a phage targeting the bacterium. In particular, Serratia marcescens , which is a pathogenic fly bacterium, can be eliminated with a phage that has been isolated from garden compost.

There are several beneficial and commercially viable insects that are affected by naturally occurring bacterial pathogens.

Experiment scheme

Isolation of specific bacteriophages from natural sources

Bacteriophages that counteract target bacteria were isolated from the initial material of the medium. Briefly, a saturated culture of Serratia marcescens was diluted in double-strength fresh tryptic soy broth (TSB) and grown for ~120 minutes to early log phase at 24-26°C or in double-strength Luria-Bertani (LB) broth and grown for ~90 min. at 37°C. Garden compost was prepared by homogenization in PBS and sterilized by filtration through a filter with a pore size of 0.2 μm. Raw waste water was sterilized by filtration through a filter with a pore size of 0.2 μm. One volume of filtered starting material was added to log-phase bacterial cultures and incubation continued for 24 hours. Enriched starting material was prepared by centrifuging the cultures and filtering the supernatant through 0.45 µm membranes.

Phages were isolated by seeding samples on bacterial lawns in a double layer of agar. Stationary bacterial cultures were pooled with LB or TSB with molten 0.6% agar and poured onto LB or TSB with 1.5% agar plates. After solidification, 2.5 μl of the diluted phage samples were applied to double agar plates and left to absorb. The cups were then wrapped and incubated overnight at 25°C (TSA) or 37°C (LB), after which they were evaluated for visible plaque formation. Freshly isolated plaques were purified by serial passage of individual plaques on the target strain by selecting plaques in SM buffer (50 mM Tris-HCl [pH 7.4], 10 mM MgSO4, 100 mM NaCl) and incubating for 15 minutes at 55°C, then repeating the agar double layering method as above using the plaque suspension.

Bacteriophages have been successfully isolated from both wastewater and compost, as detailed above. Plaque formation was clearly visible after applying the samples to S. marcescens lawns used for enrichment.

Passaging, quantification and propagation of bacteriophages

Propagation and formation of phage lysates for use in subsequent experiments was carried out using bacteriophages isolated and purified as described above. Briefly, saturated bacterial cultures were diluted 100-fold in fresh medium and grown for 60-120 minutes to reach an early log phase growth state for efficient phage infection. Phage suspensions or phage lysates were added to cultures in the early logarithmic growth phase and incubation was continued until the broth was clear, indicating phage expansion and bacterial lysis, or until 24 hours after infection. Lysates were collected by pelleting the cells by centrifugation at 7197 x g for 20 min. followed by filtration of the supernatant through membranes with a pore size of 0.45 or 0.2 μm. The filtered lysates were stored at 4°C.

The count of infectious phage particles was carried out using the method of applying to a double layer of agar. Briefly, a 1:10 serial dilution of the samples was performed in PBS and the dilutions were applied to solidified double agar plates prepared with host bacteria as described above. Plaque forming units (PFU) were counted after overnight incubation to determine the approximate titer of the samples.

In vitro analysis of isolated phages by measuring bacterial respiration

The Seahorse XFe96 analyzer (Agilent) was used to measure the effect of phages on bacteria by monitoring the oxygen consumption rate (OCR) and the extracellular acidification rate (ECAR) during infection. XFe96 plates were coated the day before experiments with 15 µl of 1 mg/ml poly-L-lysine stock solution per well and dried overnight at 28°C, and XFe96 probes were equilibrated by placing in wells containing 200 µl of XF calibrator, and incubated in the dark at room temperature. The next day, poly-L-lysine coated plates were washed twice with ddH2O. Saturated overnight cultures of E. coli BL21 (LB, 37°C) or S. marcescens (TSB, 25°C) were subcultured at 1:100 in the same medium and grown under ventilation for ~2.5 h at 30°C . The cultures were then diluted to OD 600 nm ~ 0.02 with the same medium. Processing agents were prepared by diluting stock solutions in SM buffer at 10x final concentration and loading 20 µl of 10x solutions into the appropriate sample injection ports of the probe plate. Although the probes were equilibrated in an XFe96 Flux analyzer, bacteria plates were prepared by adding 90 µl of bacterial suspensions or medium controls and centrifuged at 3000 rpm. within 10 min. After centrifugation, an additional 90 µl of appropriate medium was carefully added to the wells so as not to disturb bacterial adherence, bringing the total volume to 180 µl per well.

The XFe96 analyzer was run at ~30°C according to mix, standby, read cycles of 1:00, 0:30, 3:00. Four cycles were performed to ensure equilibration/normalization of the bacteria, then 20 μl of treatments were injected and the cycles continued as described above for a total time of approximately 6 hours. Data was analyzed using the Seahorse XFe96 Wave software package.

The effect of isolated bacteriophages was analyzed by measuring the oxygen consumption rate (OCR) and the extracellular acidification rate (ECAR) of bacteria in the Seahorse XFe96 analyzer. When E. coli was infected with T7 phage and S. marcescens was infected with freshly isolated ΦSmVL-C1 phage, significant reductions in OCR were observed after transient peaks in this rate (FIG. 4). For both phages with both hosts, the Seahorse assay allowed detection of successful phage infection without the need for plaque assays. Thus, this method is useful for detecting phage infection in a host that is not amenable to conventional phage detection methods.

Transduction assay using SYBR Gold to identify infection

Bacteriophage preparations were prepared for staining by pretreatment with nucleases to remove extraviral nucleic acids that could interfere with the interpretation of the fluorescent signal. Briefly, MgCl2 was added to 10 ml of phage lysate at a final concentration of 10 mM, and both RNase A (Qiagen) and DNase I (Sigma) were added to final concentrations of 10 μg/ml. Samples were incubated for 1 hour at room temperature. After nuclease treatment, 5 ml of lysates were combined with 1 μl of SYBR Gold (Thermo, 10000x) and incubated at room temperature for ~1.5 h. Excess dye was then removed from the samples using Amicon ultrafiltration columns. Briefly, Amicon columns (15 ml, MWCO 10 kDa) were washed by adding 10 ml of SM buffer and centrifuged at 5000 x g, 4°C for 5 minutes. Labeled phage samples were then centrifuged in columns at 5000 x g, 4°C until the volume was reduced by about 10 times (15-30 min.). To wash the samples, 5 ml of SM buffer was added to each reservoir and centrifugation was repeated followed by two additional washes. After the third wash, the remaining samples were pipetted from the Amicon reservoirs and made up to about 1 ml with SM buffer. To remove larger contaminants, washed and labeled phage samples were centrifuged at 10,000 x g for 2 minutes and supernatant samples were then filtered through 0.2 μm membranes into black microtubes and stored at 4°C.

Saturated cultures of bacteria ( E. coli MG1655 grown in LB at 37°C, S. marcescens and S. symbiotica grown in TSB at 26°C) were prepared by centrifuging 1 ml aliquots and washing once with 1 ml PBS before final resuspension. using 1 ml PBS. Positive control labeled bacteria were stained by combining 500 µl of washed bacteria and 1 µl of SYBR Gold and incubating for 1 hour in the dark at room temperature. Bacteria were pelleted by centrifugation at 8000 xg for 5 min. and washed twice with an equal volume of PBS, followed by resuspension in a final volume of 500 μl of PBS. A volume of 25 μl of stained bacteria was combined with 25 μl of SM buffer in a black microtube to which 50 μl of 10% formalin (5% final volume, ~2% formaldehyde) was added and mixed by flicking. Samples were fixed at room temperature for ~3 h and then washed with Amicon ultrafiltration columns. Briefly, 500 µl of PicoPure water was added to Amicon columns (0.5 ml, MWCO 100 kDa) and centrifuged at 14,000 xg for 5 min. for flushing membranes. Fixed samples were diluted by adding 400 µl of PBS and then transferred to pre-washed microcentrifuge columns and centrifuged at 14,000 xg for 10 min. The columns were transferred to fresh sample collection tubes and 500 µl of PBS was added to dilute the fixative remaining in the retentate. Two further dilutions were then made with PBS for a total of three washes. The final retentates were diluted to about 100 µl, then the columns were placed upside down in fresh sample collection tubes and centrifuged at 1000 xg for 2 min. to collect samples. The washed samples were transferred to black microtubes and stored at 4°C.

For transduction experiments and controls, 25 µl of bacteria (or PBS) and 25 µl of SYBR Gold labeled phage (or SM buffer) were pooled in black microtubes and incubated at rest for 15-20 min. at room temperature to ensure phage adsorption and injection into receiving bacteria. Immediately after incubation, 50 μl of 10% formalin was added to the samples and fixation was carried out at room temperature for ~4 hours. Samples were washed with PBS using Amicon columns as above.

Successful infection of a bacterial host cell required injection of bacteriophage nucleic acid. P1kc coliphage labeled with SYBR Gold and co-incubated with S. marcescens revealed the presence of fluorescent bacteria by microscopy, confirming the use of this assay in phage isolation. As with the Seahorse assay, this approach provided an alternative to traditional phage assay methods to allow extension to organisms not amenable to plaque formation assays. In addition, transduction assays using SYBR Gold did not require bacterial growth and were therefore applicable to the analysis of phages that target organisms that are difficult to culture or even impossible to culture, including endosymbionts such as Buchnera .

Phage efficacy study in vivo against S. marcescens in Drosophila melanogaster flies

Cultures of S. marcescens were grown in tryptic soy broth (TSB) at 30°C with constant shaking at 200 rpm.

The medium used to rear the initial fly populations was one containing cornmeal, molasses and yeast (11 g/l yeast, 54 g/l yellow cornmeal, 5 g/l agar, 66 ml/l molasses and 4.8 ml/l of propionic acid). All components of this diet, with the exception of propionic acid, were heated together to 80°C in deionized water with constant stirring for 30 minutes, and they were allowed to cool to 60°C. Propionic acid was then mixed in and 50 ml of the ration was aliquoted into separate vials and left to cool and solidify. Flies were reared at 26°C, 16:8 light:dark cycle, about 60% humidity.

To infect flies with S. marcescens , a fine needle (with a tip about 10 µm wide) was dipped into a solid overnight culture in stationary phase and the chest of the flies was pierced. For this experiment, four replicates of 10 males and 10 females each were infected with S. marcescens using the needle piercing method as a positive control for fly mortality. In treatment groups of four, 10 males and 10 females each were injected with S. marcescens and a solution containing approximately 108 phage particles/ml. Finally, two replicates of 10 males and 10 females each, which were not punctured or otherwise treated, were used as a negative control for mortality.

Flies in all conditions were placed in vials with food and incubated at 26°C, light:dark cycle 16:8, humidity 60%. The number of live and dead flies were counted daily for four days after piercing. All flies injected with S. marcescens only by piercing were dead within a period of 24 hours after treatment. In comparison, more than 60% of the flies in the phage treatment group and all flies in the untreated control group were alive at this time point (FIG. 5). In addition, most of the flies in the phage treatment group and the negative control group continued to survive for another four days after the experiment ended.

To determine the cause of death of the flies, dead flies from flies injected with both S. marcescens and S. marcescens +phages by piercing were homogenized and seeded. Four dead flies from each of the four replicates of both S. marcescens and S. marcescens +phage treatments were homogenized in 100 μl TSB. A 1:100 dilution was also made by diluting the homogenate in TSB. 10 μl of the concentrated homogenate as well as a 1:100 dilution were plated onto TSA plates and incubated overnight at 30°C. When examining plates for bacterial growth, all plates of dead flies injected with S. marcescens by piercing had a lawn of bacteria growing on them, while plates of dead flies injected with S. marcescens + phages by piercing did not it was bacteria. This indicated that, in the absence of the phage, S. marcescens probably induced septic shock in the flies, leading to their death. At the same time, in the presence of phage, mortality could be due to damage caused by needle piercing.

OTHER IMPLEMENTATION OPTIONS

Although the foregoing invention has been described in sufficient detail by means of illustrations and examples for the purposes of clarity of understanding, the descriptions and examples should not be construed as limiting the scope of the present invention. The disclosures of all patent and scientific literature sources cited herein are expressly incorporated in their entirety by reference.

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SEQUENCE LIST

<110> Flagship Pioneering, Inc.

<120> METHODS AND RELATED COMPOSITIONS FOR MANUFACTURING PRODUCTS

FOOD AND FEED

<130> 51215-007WO3

<150> US 62/584,011

<151> 2017-11-09

<150> US 62/450,038

<151> 2017-01-24

<160> 228

<170> PatentIn version 3.5

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<212> DNA

<213> Carsonella ruddii

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gtgtacaagg ctcgagaacg tattcaccgt aacattctga tttacgatta ctagcgattc 180

240

cattatgttg ccatatagca tataactttt tgtaatactc attgtagcac gtgtgtagcc 300

ctacttataa gggccatgat gacttgacgt cgtcctcacc ttcctccaat ttatcattgg 360

420

aggacttaac ccaacatttc acaacacgag ctgacgacag ccatgcagca cctgtctcaa 480

agctaaaaaa gctttattat ttctaataaa ttctttggat gtcaaaagta ggtaagattt 540

ttcgtgttgt atcgaattaa accacatgct ccaccgcttg tgcgagcccc cgtcaattca 600

tttgagtttt aaccttgcgg tcgtaatccc caggcggtca acttaacgcg ttagcttttt 660

cactaaaaat atataacttt ttttcataaa acaaaattac aattataata tttaataaat 720

agttgacatc gtttactgca tggactacca gggtatctaa tcctgtttgc tccccatgct 780

ttcgtgtatt agtgtcagta ttaaaataga aatacgcctt cgccactagt attctttcag 840

atatctaagc atttcactgc tactcctgaa attctaattt cttcttttat actcaagttt 900

ataagtatta atttcaatat taaattactt taataaattt aaaaattaat ttttaaaaac 960

aacctgcaca ccctttacgc ccaataattc cgattaacgc ttgcacccct cgtattaccg 1020

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360

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<213> Buchnera aphidicola, strain APS (Acyrthosiphon pisum)

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660

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<213> Buchnera aphidicola strain Sg (Schizaphis graminum)

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ggagcaaaca ggattagata ccctggtagt ccatgccgta aacgatgtcg acttggaggt 840

tgtttccaag agaagtgact tccgaagcta acgcgttaag tcgaccgcct ggggagtacg 900

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aataaaaaag tgccttcggg aactgtgaga caggtgctgc atggctgtcg tcagctcgtg 1080

1140

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1380

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<212> DNA

<213> Buchnera aphidicola strain Bp (Baizongia pistaciae)

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ggaggcagca gtggggaata ttgcacaatg ggctaaagcc tgatgcagct atgccgcgtg 420

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gcggtaatac ggagggtgcg agcgttaatc agaattactg ggcgtaaaga gcacgtaggc 600

ggtttattaa gtcagatgtg aaatccctag gcttaactta ggaactgcat ttgaaactaa 660

tagactagag tctcatagag ggaggtagaa ttctaggtgt agcggtgaaa tgcgtagata 720

tctagaggaa tacccgtggc gaaagcgacc tcctaaatga aaactgacgc tgaggtgcga 780

aagcgtgggg agcaaacagg attagatacc ctggtagtcc atgctgtaaa cgatgtcgac 840

ttggaggttg tttcctagag aagtggcttc cgaagctaac gcattaagtc gaccgcctgg 900

ggagtacggt cgcaaggcta aaactcaaat gaattgacgg gggcccgcac aagcggtgga 960

gcatgtggtt taattcgatg caacgcgaag aaccttacct ggtcttgaca tccatagaat 1020

tttttagaga taaaagagtg ccttagggaa ctatgagaca ggtgctgcat ggctgtcgtc 1080

agctcgtgtt gtgaaatgtt gggttaagtc ccgcaacgag cgcaacccct atcctttgtt 1140

gccatcaggt tatgctggga actcagagga gactgccggt tataaaccgg aggaaggtgg 1200

ggatgacgtc aagtcatcat ggcccttacg accagggcta cacacgtgct acaatggcat 1260

atacaaagag atgcaactct gcgaagataa gcaaacctca taaagtatgt cgtagtccgg 1320

actggagtct gcaactcgac tccacgaagt aggaatcgct agtaatcgtg gatcagaatg 1380

ccacggtgaa tacgttcccg ggccttgtac acaccgcccg tcacaccatg ggagtggggtt 1440

gcaaaagaag caggtagctt aaccagatta ttttattgga gggcgcttac cactttgtga 1500

ttcatgactg gggtgaagtc gtaacaaggt aaccgtaggg gaacctgcgg ttggatcacc 1560

tccta 1566

<210> 6

<211> 828

<212> DNA

<213> Buchnera aphidicola BCc

<400> 6

atgagatcat taatatataa aaatcatgtt ccaattaaaa aattaggaca aaatttttta 60

cagaataaag aaattattaa tcagataatt aatttaataa atattaataa aaatgataat 120

attattgaaa taggatcagg attaggagcg ttaacttttc ctatttgtag aatcattaaa 180

240

attaaaaaat tacaaattat aattgctgat

tctttacaga aatataaaaa atataggtttt attggtaatt taccatataa tattgctact 360

atattttttt taaaaacaat taaatttctt tataatataa ttgatatgca ttttatgttt 420

caaaaagaag tagcaaagag attattagct actcctggta ctaaagaata tggtagatta 480

agtattattg cacaatattt ttataagata gaaactgtta ttaatgttaa taaatttaat 540

ttttttccta ctcctaaagt agattctact ttttttacgat ttactcctaa atattttaat 600

agtaaatata aaatagataa acatttttct gttttagaat taattactag attttctttt 660

caacatagaa gaaaattttt aaataataat ttaatatctt tattttctac aaaagaatta 720

atttctttag atattgatcc atattcaaga gcagaaaatg tttctttaat tcaatattgt 780

aaattaatga aatattattt gaaaagaaaa attttatgtt tagattaa 828

<210> 7

<211> 921

<212> DNA

<213> Buchnera aphidicola (Cinara tujafilina)

<400> 7

ttatcttatt tcacatatac gtaatattgc gctgcgtgca cgaggatttt tttgaatttc 60

120

180

gttttttaca atcctatctt caatagaatg aaaactaata atagcaattt ttgatccgga 240

atgtaatatg ttaataataa tttttaatat tttatgtaat tcatttattt cttggttaat 300

atatattcga aaagcttgaa atgttctcgt agctggatgt ttaaatttgt catattttgg 360

420

ttgtaatatg atggctcggg atattttttt tgcgtatttt tcttcgccaa aattttttat 480

tacctgttct attgtttttt ggtttgtttt ttttaaccat tgactaactg atattccaga 540

tttagggttc atacgcatat ctaaaggtcc atcattcata aatgaaaatc ctcggatact 600

agaatttaac tgtattgaag aaatacctaa atctaataat attccatcta ttttatctct 660

atttttttct ttttttaata ttttttcaat attagaaaat ttacctaaaa atattttaaa 720

tcgcgaatct tttatttttt ttccgatttt tatagattgt gggtcttgat caatactata 780

taactttcca ttaaccccta attcttgaag aattgctttt gaatgaccac cacctccaaa 840

tgtacaatca acatatgtac cgtctttttt tatttttaag tattgtatga tttcttttgt 900

taaaacaggt ttatgaatca t 921

<210> 8

<211> 822

<212> DNA

<213> Buchnera aphidicola strain G002 (Myzus persicae)

<400> 8

atgaaaagta taaaaacttt taaaaaacac ttttcctgtga aaaaatatgg acaaaatttt

cttattaata aagagatcat aaaaaatatt gttaaaaaaa ttaatccaaa tatagaacaa 120

acattagtag aaatcggacc aggattagct gcattaactg agcccatatc tcagttatta 180

aaagagttaa tagttattga aatagactgt aatctattat attttttaaa aaaacaacca 240

ttttattcaa aattaatagt tttttgtcaa gatgctttaa actttaatta tacaaattta 300

ttttataaaa aaaataaatt aattcgtatt ttttggtaatt taccatataa tatctctaca 360

420

caaaaagaag ttgctgcaag attaattgca ttacctggaa ataaatatta cggtcgtttg 480

agcattatat ctcaatatta ttgtgatatc aaaattttat taaatgttgc tcctgaagat 540

ttttggccta ttccgagagt tcattctata tttgtaaatt taacacctca tcataattct 600

660

agaagaaaaa tattacgtca tagtttaaaa aatttatttt ctgaaacaac tttattaaat 720

780

gctaattatt tgtataaaaa aaattatact aaaaaaaatt aa 822

<210> 9

<211> 822

<212> DNA

<213> Buchnera aphidicola strain Ak (Acyrthosiphon kondoi)

<400> 9

attataaaaa attttaaaaa acattttcct ttaaaaaggt atggacaaaa ttttcttgtc 60

aatacaaaaa ctattcaaaa gataattaat ataattaatc caaacaccaa acaaacatta 120

gtggaaattg gacctggatt agctgcatta acaaaaccaa tttgtcaatt attagaagaa 180

240

300

360

attatttctt tattcaatca tattaaagtt attcaagata tgaattttat gttacagaaa 420

gaggttgctg aaagattaat ttctattcct ggaaataaat cttatggccg tttaagcatt 480

atttctcagt attattgtaa aattaaaata ttattaaatg ttgtacctga agattttcga 540

cctataccga aagtgcattc tgtttttatc aatttaactc ctcataccaa ttctccatat 600

tttgttttg atacaaatat cctcagttct atcacaagaa atgcttttca aaatagaagg 660

aaaattttgc gtcatagttt aaaaaattta ttttctgaaa aagaactaat tcaattagaa 720

attaatccaa atttacgagc tgaaaatatt tctatctttc agtattgtca attagctgat 780

tatttatata aaaaattaaa taatcttgta aaaatcaatt aa 822

<210> 10

<211> 822

<212> DNA

<213> Buchnera aphidicola strain Ua (Uroleucon ambrosiae)

<400> 10

60

gtaaatagag aaataatcaa aaatattatc aaaataatta atcctaaaaa aacgcaaaca 120

ttattagaaa ttggaccggg tttaggtgcg ttaacaaaac ctatttgtga atttttaaat 180

240

tttgataaat taaaaatata ttgtcataat gctttagatt ttaattataa aaatatttc 300

tataaaaaaa gtcaattaat tcgtattttt ggaaatttac catataatat ttctacatct 360

420

caagaagtgg ctaaaagatt agttgctatt cctggtgaaa aactttatgg tcgtttaagt 480

attatatctc aatattattg taatattaaa atattattac atattcgacc tgaaaatttt 540

caacctattc ctaaagttaa ttcaatgttt gtaaatttaa ctccgcatat tcattctcct 600

tattttgttt atgatattaa ttttattaact agtattacaa aacatgcttt tcaacataga 660

agaaaaatat tgcgtcatag tttaagaaat ttttttttctg agcaagattt aattcattta 720

gaaattaatc caaatttaag agctgaaaat gtttctatta ttcaatattg tcaattggct 780

aataatttat ataaaaaaca taaacagttt attaataatt aa 822

<210> 11

<211> 816

<212> DNA

<213> Buchnera aphidicola (Aphis glycines)

<400> 11

60

attaaaaaaa taattaaatt tattaatccg cagttaaatg aaatattggt tgaaattgga 120

ccgggattag ctgctatcac tcgacctatt tgtgatttga tagatcattt aattgtgatt 180

gaaattgata aaattttatt agatagatta aaacagttct cattttattc aaaattaaca 240

gtatatcatc aagatgcttt agcatttgat tacataaagt tatttaataa aaaaaataaa 300

360

tttaaaagaa ttaatattat taaagatatg aattttatgc tacaaaaaga agttgctgaa 420

tagtaaatta tatggtcgtt taagtattat ttctcaatat 480

tattgtaata taaaagtttt attgcatgtg tcttcaaaat gttttaaacc agttcctaaa 540

600

aatgtaaacg ttcttagtta tattacaaat ttagcttttc aaaaaagaag aaaaatatta 660

cgtcatagtt taggtaaaat attttctgaa aaagttttta taaaattaaa tattaatccc 720

aaattaagac ctgagaatat ttctatatta caatattgtc agttatctaa ttatatgata 780

gaaaataata ttcatcagga acatgtttgt atttaa 816

<210> 12

<211> 1463

<212> DNA

<213> Annandia pinicola

<400> 12

agattgaacg ctggcggcat gccttacaca tgcaagtcga acggtaacag gtcttcggac 60

gctgacgagt ggcgaacggg tgagtaatac atcggaacgt gcccagtcgt gggggataac 120

tactcgaaag agtagctaat accgcatacg atctgaggat gaaagcgggg gaccttcggg 180

cctcgcgcga ttggagcggc cgatggcaga ttaggtagtt ggtgggataa aagcttacca 240

agccgacgat ctgtagctgg tctgagagga cgaccagcca cactggaact gagatacggt 300

cggactctt acgggaggca gcagtggggga atattgcaca atgggcgcaa gcctgatgca 360

gctatgtcgc gtgtatgaag aagaccttag ggttgtaaag tactttcgat agcataagaa 420

gataatgaga ctaataattt tattgtctga cgttagctat agaagaagca ccggctaact 480

ccgtgccagc agccgcggta atacgggggg tgctagcgtt aatcggaatt actgggcgta 540

aagagcatgt aggtggttta ttaagtcaga tgtgaaatcc ctggacttaa tctaggaact 600

gcatttgaaa ctaataggct agagtttcgt agagggaggt agaattctag gtgtagcggt 660

gaaatgcata gatatctaga ggaatatcag tggcgaaggc gaccttctgg acgataactg 720

acgctaaaat gcgaaagcat gggtagcaaa caggattaga taccctggta gtccatgctg 780

taaacgatgt cgactaagag gttggaggta taacttttaa tctctgtagc taacgcgtta 840

agtcgaccgc ctggggagta cggtcgcaag gctaaaactc aaatgaattg acgggggcct 900

gcacaagcgg tggagcatgt ggtttaattc gatgcaacgc gtaaaacctt acctggtctt 960

gacatccaca gaattttaca gaaatgtaga agtgcaattt gaactgtgag acaggtgctg 1020

catggctgtc gtcagctcgt gttgtgaaat gttgggttaa gtcccgcaac gagcgcaacc 1080

cttgtccttt gttaccataa gatttaagga actcaaagga gactgccggt gataaactgg 1140

aggaaggcgg ggacgacgtc aagtcatcat ggcccttatg accagggcta cacacgtgct 1200

acaatggcat atacaaagag atgcaatatt gcgaaataaa gccaatctta taaaatatgt 1260

cctagttcgg actggagtct gcaactcgac tccacgaagt cggaatcgct agtaatcgtg 1320

1380

gaagtggatt gcaaaagaag taagaaaatt aaccttctta acaaggaaat aacttaccac 1440

tttgtgactc ataactgggg tga 1463

<210> 13

<211> 1554

<212> DNA

<213> Moranella endobia

<400> 13

tctttttggt aaggaggtga tccaaccgca ggttccccta cggttacctt gttacgactt 60

caccccagtc atgaatcaca aagtggtaag cgccctccta aaaggttagg ctacctactt 120

cttttgcaac ccacttccat ggtgtgacgg gcggtgtgta caaggcccgg gaacgtattc 180

accgtggcat tctgatccac gattactagc gattcctact tcatggagtc gagttgcaga 240

ctccaatccg gactacgacg cactttatga ggtccgctaa ctctcgcgag cttgcttctc 300

tttgtatgcg ccattgtagc acgtgtgtag ccctactcgt aagggccatg atgacttgac 360

gtcatcccca ccttcctccg gtttatcacc ggcagtctcc tttgagttcc cgaccgaatc 420

gctggcaaaa aaggataagg gttgcgctcg ttgcgggact taacccaaca tttcacaaca 480

cgagctgacg acagccatgc agcacctgtc tcagagttcc cgaaggtacc aaaacatctc 540

tgctaagttc tctggatgtc aaggtaggt aaggttcttc gcgttgcatc gaattaaacc 600

acatgctcca ccgcttgtgc gggcccccgt caattcattt gagttttaac cttgcggccg 660

tactccccag gcggtcgatt taacgcgtta actacgaaag ccacagttca agaccacagc 720

tttcaaatcg acatagttta cggcgtggac taccagggta tctaatcctg tttgctcccc 780

acgctttcgt acctgagcgt cagtattcgt ccagggggcc gccttcgcca ctggtattcc 840

tccagatatc tacacatttc accgctacac ctggaattct acccccctct acgagactct 900

agcctatcag tttcaaatgc agttcctagg ttaagcccag ggatttcaca tctgacttaa 960

taaaccgcct acgtactctt tacgcccagt aattccgatt aacgcttgca ccctccgtat 1020

taccgcggct gctggcacgg agttagccgg tgcttcttct gtaggtaacg tcaatcaata 1080

accgtattaa ggatattgcc ttcctcccta ctgaaagtgc tttacaaccc gaaggccttc 1140

ttcacacacg cggcatggct gcatcagggt ttcccccatt gtgcaatatt ccccactgct 1200

gcctcccgta ggagtctgga ccgtgtctca gttccagtgt ggctggtcat cctctcagac 1260

cagctaggga tcgtcgccta ggtaagctat tacctcacct actagctaat cccatctggg 1320

ttcatctgaa ggtgtgaggc caaaaggtcc cccactttgg tcttacgaca ttatgcggta 1380

ttagctaccg tttccagcag ttatccccct ccatcaggca gatccccaga ctttactcac 1440

ccgttcgctg ctcgccggca aaaaagtaaa ctttttttccg ttgccgctca acttgcatgt 1500

gttaggcctg ccgccagcgt tcaatctgag ccatgatcaa actcttcaat taaa 1554

<210> 14

<211> 1539

<212> DNA

<213> Ishikawaella capsulata Mpkobe

<400> 14

aaattgaaga gtttgatcat ggctcagatt gaacgctagc ggcaagctta acacatgcaa 60

gtcgaacggt aacagaaaaa agcttgcttt tttgctgacg agtggcggac gggtgagtaa 120

tgtctgggga tctacctaat ggcgggggat aactactgga aacggtagct aataccgcat 180

aatgttgtaa aaccaaagtg ggggacctta tggcctcaca ccattagatg aacctagatg 240

ggattagctt gtaggtgggg taaaggctca cctaggcaac gatccctagc tggtctgaga 300

ggatgaccag ccacactgga actgagatac ggtccagact cctacgggag gcagcagtgg 360

ggaatcttgc acaatgggcg caagcctgat gcagctatgt cgcgtgtatg aagaaggcct 420

tagggttgta aagtactttc atcggggaag aaggatatga gcctaatatt ctcatatatt 480

gacgttacct gcagaagaag caccggctaa ctccgtgcca gcagccgcgg taacacggag 540

ggtgcgagcg ttaatcggaa ttactgggcg taaagagcac gtaggtggtt tattaagtca 600

tatgtgaaat ccctgggctt aacctaggaa ctgcatgtga aactgataaa ctagagtttc 660

gtagagggag gtggaattcc aggtgtagcg gtgaaatgcg tagatatctg gaggaatatc 720

agaggcgaag gcgaccttct ggacgaaaac tgacactcag gtgcgaaagc gtggggagca 780

aacagatta gataccctgg tagtccacgc tgtaaacaat gtcgactaaa aaactgtgag 840

cttgacttgt ggtttttgta gctaacgcat taagtcgacc gcctggggag tacggccgca 900

aggttaaaac tcaaatgaat tgacggggggt ccgcacaagc ggtggagcat gtggtttaat 960

tcgatgcaac gcgaaaaacc ttacctggtc ttgacatcca gcgaattata tagaaatata 1020

taagtgcctt tcggggaact ctgagacgct gcatggctgt cgtcagctcg tgttgtgaaa 1080

1140

actcagagga gactgccagt attaaactgg aggaaggtgg ggatgacgtc aagtcatcat 1200

ggcccttatg accagggcta cacacgtgct acaatggtgt atacaaagag aagcaatctc 1260

gcaaagtaa gcaaaactca aaaagtacat cgtagttcgg attagagtct gcaactcgac 1320

tctatgaagt aggaatcgct agtaatcgtg gatcagaatg ccacggtgaa tacgttctct 1380

ggccttgtac acaccgcccg tcacaccatg ggagtaagtt gcaaaagaag taggtagctt 1440

aacctttata ggagggcgct taccactttg tgatttatga ctggggtgaa gtcgtaacaa 1500

ggtaactgta gggggaacctg tggttggatt acctcctta 1539

<210> 15

<211> 1561

<212> DNA

<213> Baumannia cicadellinicola

<400> 15

ttcaattgaa gagtttgatc atggctcaga ttgaacgctg gcggtaagct taacacatgc 60

aagtcgagcg gcatcggaaa gtaaattaat tactttgccg gcaagcggcg aacgggtgag 120

taatatctgg ggatctacct tatggagagg gataactatt ggaaacgata gctaacaccg 180

cataatgtcg tcagaccaaa atgggggacc taatttaggc ctcatgccat aagatgaacc 240

cagatgagat tagctagtag gtgagataat agctcaccta ggcaacgatc tctagttggt 300

ctgagaggat gaccagccac actggaactg agacacggtc cagactccta cgggaggcag 360

cagtggggaa tcttgcacaa tgggggaaac cctgatgcag ctataccgcg tgtgtgaaga 420

aggccttcgg gttgtaaagc actttcagcg gggaagaaaa tgaagttact aataataatt 480

gtcaattgac gttacccgca aaagaagcac cggctaactc cgtgccagca gccgcggtaa 540

600 gacggagggt gcaagcgtta

ttagtcaggt gtgaaagccc taggcttaac ctaggaattg catttgaaac tggtaagcta 660

gagtctcgta gaggggggga gaattccagg tgtagcggtg aaatgcgtag agatctggaa 720

gaataccagt ggcgaaggcg cccccctgga cgaaaactga cgctcaagta cgaaagcgtg 780

gggagcaaac aggattagat accctggtag tccacgctgt aaacgatgtc gatttgaagg 840

ttgtagcctt gagctatagc tttcgaagct aacgcattaa atcgaccgcc tggggagtac 900

gaccgcaagg ttaaaactca aatgaattga cgggggcccg cacaagcggt ggagcatgtg 960

gtttaattcg atacaacgcg aaaaacctta cctactcttg acatccagag tataaagcag 1020

aaaagcttta gtgccttcgg gaactctgag acaggtgctg catggctgtc gtcagctcgt 1080

1140

attaagtcgg gaactcaaag gagactgccg gtgataaacc ggaggaaggt gaggataacg 1200

tcaagtcatc atggccctta cgagtagggc tacacacgtg ctacaatggt gcatacaaag 1260

agaagcaatc tcgtaagagt tagcaaacct cataaagtgc atcgtagtcc ggattagagt 1320

ctgcaactcg actctatgaa gtcggaatcg ctagtaatcg tggatcagaa tgccacggtg 1380

aatacgttcc cgggccttgt acacaccgcc cgtcacacca tgggagtgta ttgcaaaaga 1440

agttagtagc ttaactcata atacgagagg gcgcttacca ctttgtgatt cataactggg 1500

gtgaagtcgt aacaaggtaa ccgtagggga acctgcggtt ggatcacctc cttacactaa 1560

a 1561

<210> 16

<211> 1464

<212> DNA

<213> Sodalis-like

<400> 16

attgaacgct ggcggcaggc ctaacacatg caagtcgagc ggcagcggga agaagcttgc 60

ttctttgccg gcgagcggcg gacgggtgag taatgtctgg ggatctgccc gatggagggg 120

gtaactact ggaaacggta gctaataccg cataacgtcg caagaccaaa gtgggggacc 180

ttcggggcctc acaccatcgg atgaacccag gtgggattag ctagtaggtg gggtaatggc 240

tcacctaggc gacgatccct agctggtctg agaggatgac cagtcacact ggaactgaga 300

cacggtccag actcctacgg gaggcagcag tggggaatat tgcacaatgg gggaaaccct 360

gatgcagcca tgccgcgtgt gtgaagaagg ccttcgggtt gtaaagcact ttcagcgggg 420

aggaaggcga tggcgttaat agcgctatcg attgacgtta cccgcagaag aagcaccggc 480

taactccgtg ccagcagccg cggtaatacg gagggtgcga gcgttaatcg gaattactgg 540

gcgtaaagcg tacgcaggcg gtctgttaag tcagatgtga aatcccgggg ctcaacctgg 600

gaactgcatt tgaaactggc aggctagagt ctcgtagagg ggggtagaat tccaggtgta 660

gcggtgaaat gcgtagagat ctggaggaat accggtggcg aaggcggccc cctggacgaa 720

gactgacgct caggtacgaa agcgtgggga gcaaacagga ttagataccc tggtagtcca 780

cgctgtaaac gatgtcgatt tgaaggttgt ggccttgagc cgtggctttc ggagctaacg 840

tgttaaatcg accgcctggg gagtacggcc gcaaggttaa aactcaaatg aattgacggg 900

ggccgcaca agcggtggag catgtggttt aattcgatgc aacgcgaaga accttaccta 960

1020

gtgctgcatg gctgtcgtca gctcgtgttg tgaaatgttg ggttaagtcc cgcaacgagc 1080

1140

taaaccggag gaaggtgggg atgacgtcaa gtcatcatgg cccttacgag tagggctaca 1200

cacgtgctac aatggcgcat acaaagagaa gcgatctcgc gagagtcagc ggacctcata 1260

aagtgcgtcg tagtccggat tggagtctgc aactcgactc catgaagtcg gaatcgctag 1320

taatcgtgga tcagaatgcc acggtgaata cgttcccggg ccttgtacac accgcccgtc 1380

acaccatggg agtgggttgc aaaagaagta ggtagcttaa ccttcgggag ggcgcttacc 1440

actttgtgat tcatgactgg ggtg 1464

<210> 17

<211> 1465

<212> DNA

<213> Hartigia pinicola

<400> 17

agatttaacg ctggcggcag gcctaacaca tgcaagtcga gcggtaccag aagaagcttg 60

cttcttgctg acgagcggcg gacgggtgag taatgtatgg ggatctgccc gacagagggg 120

gtaactatt ggaaacggta gctaataccg cataatctct gaggagcaaa gcaggggaac 180

ttcggtcctt gcgctatcgg atgaacccat atgggattag ctagtaggtg aggtaatggc 240

tcccctaggc aacgatccct agctggtctg agaggatgat cagccacact gggactgaga 300

cacggcccag actcctacgg gaggcagcag tggggaatat tgcacaatgg gcgaaagcct 360

gatgcagcca tgccgcgtgt atgaagaagg ctttagggtt gtaaagtact ttcagtcgag 420

aggaaaacat tgatgctaat atcatcaatt attgacgttt ccgacagaag aagcaccggc 480

taactccgtg ccagcagccg cggtaatacg gagggtgcaa gcgttaatcg gaattactgg 540

gcgtaaagcg cacgcaggcg gttaattaag ttagatgtga aagccccgggg cttaacccag 600

gaatagcata taaaactggt caactagagt attgtagagg ggggtagaat tccatgtgta 660

gcggtgaaat gcgtagagat gtggaggaat accagtggcg aaggcggccc cctggacaaa 720

aactgacgct caaatgcgaa agcgtgggga gcaaacagga ttagataccc tggtagtcca 780

tgctgtaaac gatgtcgatt tggaggttgt tcccttgagg agtagcttcc gtagctaacg 840

cgttaaatcg accgcctggg ggagtacgac tgcaaggtta aaactcaaat gaattgacgg 900

gggcccgcac aagcggtgga gcatgtggtt taattcgatg caacgcgaaa aaccttacct 960

actcttgaca tccagataat ttagcagaaa tgctttagta ccttcgggaa atctgagaca 1020

ggtgctgcat ggctgtcgtc agctcgtgtt gtgaaatgtt gggttaagtc ccgcaacgag 1080

cgcaaccctt atcctttgtt gccagcgatt aggtcgggaa ctcaaaggag actgccggtg 1140

ataaaccgga ggaaggtggg gatgacgtca agtcatcatg gcccttacga gtagggctac 1200

acacgtgcta caatggcata tacaaaggga agcaacctcg cgagagcaag cgaaactcat 1260

aaattatgtc gtagttcaga ttggagtctg caactcgact ccatgaagtc ggaatcgcta 1320

gtaatcgtag atcagaatgc tacggtgaat acgttcccgg gccttgtaca caccgcccgt 1380

cacaccatgg gagtgggttg caaaagaagt aggtaactta accttatgga aagcgcttac 1440

cactttgtga ttcataactg gggtg 1465

<210> 18

<211> 1571

<212> DNA

<213> Tremblaya phenacola

<400> 18

aggtaatcca gccacacctt ccagtacggc taccttgtta cgacttcacc ccagtcacaa 60

cccttacctt cggaactgcc ctcctcacaa ctcaaaccac caaacacttt taaatcaggt 120

tgagagaggt taggcctgtt acttctggca agaattattt ccatggtgtg acgggcggtg 180

tgtacaagac ccgagaacat attcaccgtg gcatgctgat ccacgattac tagcaattcc 240

aacttcatgc actcgagttt cagagtacaa tccgaactga ggccggcttt gtgagattag 300

ctcccttttg caagttggca actctttggt ccggccattg tatgatgtgt gaagccccac 360

ccataaaggc catgaggact tgacgtcatc cccaccttcc tccaacttat cgctggcagt 420

ctctttaagg taactgacta atccagtagc aattaaagac aggggttgcg ctcgttacag 480

gacttaaccc aacatctcac gacacgagct gacgacagcc atgcagcacc tgtgcactaa 540

ttctctttca agcactcccg cttctcaaca ggatcttagc catatcaaag gtaggtaagg 600

tttttcgcgt tgcatcgaat taatccacat catccactgc ttgtgcgggt ccccgtcaat 660

tcctttgagt tttaaccttg cggccgtact ccccaggcgg tcgacttgtg cgttagctgc 720

accactgaaa aggaaaactg cccaatggtt agtcaacatc gtttagggca tggactacca 780

gggtatctaa tcctgtttgc tccccatgct ttagtgtctg agcgtcagta acgaaccagg 840

aggctgccta cgctttcggt attcctccac atctctacac atttcactgc tacatgcgga 900

attctacctc cccctctcgt actccagcct gccagtaact gccgcattct gaggttaagc 960

ctcagccttt cacagcaatc ttaacaggca gcctgcacac cctttacgcc caataaatct 1020

gattaacgct cgcaccctac gtattaccgc ggctgctggc acgtagtttg ccggtgctta 1080

ttctttcggt acagtcacac caccaaattg ttagttggggt ggctttcttt ccgaacaaaa 1140

gtgctttaca acccaaaggc cttcttcaca cacgcggcat tgctggatca ggcttccgcc 1200

cattgtccaa gattcctcac tgctgccttc ctcagaagtc tgggccgtgt ctcagtccca 1260

gtgtggctgg ccgtcctctc agaccagcta ccgatcattg ccttgggaag ccattacctt 1320

tccaacaagc taatcagaca tcagccaatc tcagagcgca aggcaattgg tcccctgctt 1380

tcattctgct tggtagagaa ctttatgcgg tattaattag gctttcacct agctgtcccc 1440

cactctgagg catgttctga tgcattactc acccgtttgc cacttgccac caagcctaag 1500

cccgtgttgc cgttcgactt gcatgtgtaa ggcatgccgc tagcgttcaa tctgagccag 1560

gatcaaactc t 1571

<210> 19

<211> 1535

<212> DNA

<213> Tremblaya princeps

<400> 19

agagtttgat cctggctcag attgaacgct agcggcatgc attacacatg caagtcgtac 60

ggcagcacgg gcttaggcct ggtggcgagt ggcgaacggg tgagtaacgc ctcggaacgt 120

gccttgtagt gggggatagc ctggcgaaag ccagattaat accgcatgaa gccgcacagc 180

atgcgcggtg aaagtggggg attctagcct cacgctactg gatcggccgg ggtctgatta 240

gctagttggc ggggtaatgg cccaccaagg cttagatcag tagctggtct gagaggacga 300

tcagccacac tgggactgag acacggccca gactcctacg ggaggcagca gtggggaatc 360

ttggacaatg ggcgcaagcc tgatccagca atgccgcgtg tgtgaagaag gccttcgggt 420

cgtaaagcac ttttgttcgg gatgaagggg ggcgtgcaaa caccatgccc tcttgacgat 480

accgaaagaa taagcaccgg ctaactacgt gccagcagcc gcggtaatac gtagggtgcg 540

agcgttaatc ggaatcactg ggcgtaaagg gtgcgcgggt ggtttgccaa gacccctgta 600

aaatcctacg gcccaaccgt agtgctgcgg aggttactgg taagcttgag tatggcagag 660

gggggtagaa ttccaggtgt agcggtgaaa tgcgtagata tctggaggaa taccgaaggc 720

gaaggcaacc ccctgggcca tcactgacac tgaggcacga aagcgtgggg agcaaacagg 780

attagatacc ctggtagtcc acgccctaaa ccatgtcgac tagttgtcgg ggggagccct 840

ttttcctcgg tgacgaagct aacgcatgaa gtcgaccgcc tggggagtac gaccgcaagg 900

ttaaaactca aaggaattga cggggacccg cacaagcggt ggatgatgtg gattaattcg 960

atgcaacgcg aaaaacctta cctacccttg acatggcgga gattctgccg agaggcggaa 1020

gtgctcgaaa gagaatccgt gcacaggtgc tgcatggctg tcgtcagctc gtgtcgtgag 1080

atgttgggtt aagtcccata acgagcgcaa cccccgtctt tagttgctac cactggggca 1140

ctctatagag actgccggtg ataaaccggga ggaaggtggg gacgacgtca agtcatcatg 1200

gcctttatgg gtagggcttc acacgtcata caatggctgg agcaaagggt cgccaactcg 1260

agagagggag ctaatcccac aaacccagcc ccagttcgga ttgcactctg caactcgagt 1320

gcatgaagtc ggaatcgcta gtaatcgtgg atcagcatgc cacggtgaat acgttctcgg 1380

gtcttgtaca caccgcccgt cacaccatgg gagtaagccg catcagaagc agcctcccta 1440

accctatgct gggaaggagg ctgcgaaggt ggggtctatg actggggtga agtcgtaaca 1500

aggtagccgt accggaaggt gcggctggat tacct 1535

<210> 20

<211> 1450

<212> DNA

<213> Nasuia deltocephalinicola

<400> 20

agtttaatcc tggctcagat ttaacgcttg cgacatgcct aacacatgca agttgaacgt 60

tgaaaatatt tcaaagtagc gtataggtga gtataacatt taaacatacc ttaaagttcg 120

gaataccccg atgaaaatcg gtataatacc gtataaaagt atttaagaat taaagcgggg 180

aaaacctcgt gctataagat tgttaaatgc ctgattagtt tgttggtttt taaggtaaaa 240

gcttaccaag actttgatca gtagctattc tgtgaggatg tatagccaca ttgggattga 300

aataatgccc aaacctctac ggagggcagc agtggggaat attggacaat gagcgaaagc 360

ttgatccagc aatgtcgcgt gtgcgattaa gggaaactgt aaagcacttt tttttaagaa 420

taagaaattt taattaataa ttaaaatttt tgaatgtatt aaaagaataa gtaccgacta 480

atcacgtgcc agcagtcgcg gtaatacgtg gggtgcgagc gttaatcgga tttattgggc 540

gtaaagtgta ttcaggctgc ttaaaaagat ttatattaaa tatttaaatt aaatttaaaa 600

660

agtgaaatgc gttgatatat aaaggaacgc cgaaagcgaa agcatttttc tgtaatagaa 720

ctgacgctta tatacgaaag cgtgggtagc aaacaggatt agataccctg gtagtccacg 780

ccctaaacta tgtcaattaa ctattagaat tttttttagt ggtgtagcta acgcgttaaa 840

ttgaccgcct gggtattacg atcgcaagat taaaactcaa aggaattgac ggggaccagc 900

acaagcggtg gatgatgtgg attaattcga tgatacgcga aaaaccttac ctgcccttga 960

catggttaga attttattga aaaataaaag tgcttggaaa agagctaaca cacaggtgct 1020

1080

ccctactctt agttgctaat taaagaactt taagagaaca gctaacaata agtttagagg 1140

aaggagggga tgacttcaag tcctcatggc ccttatgggc agggcttcac acgtcataca 1200

atggttaata caaaaagttg caatatcgta agattgagct aatctttaaa attaatctta 1260

gttcggattg tactctgcaa ctcgagtaca tgaagttgga atcgctagta atcgcggatc 1320

agcatgccgc ggtgaatagt ttaactggtc ttgtacacac cgcccgtcac accatggaaa 1380

taaatcttgt tttaaatgaa gtaatatatt ttatcaaaac aggttttgta accggggtga 1440

agtcgtaaca 1450

<210> 21

<211> 1536

<212> DNA

<213> Zinderia insecticola CARI

<400> 21

atataaataa gagtttgatc ctggctcaga ttgaacgcta gcggtatgct ttacacatgc 60

aagtcgaacg acaatattaa agcttgcttt aatataaagt ggcgaacggg tgagtaatat 120

atcaaaacgt accttaaagt gggggataac taattgaaaa attagataat accgcatatt 180

aatcttagga tgaaaatagg aataatatct tatgctttta gatcggttga tatctgatta 240

gctagttggt agggtaaatg cttaccaagg caatgatcag tagctggttt tagcgaatga 300

tcagccacac tggaactgag acacggtcca gacttctacg gaaggcagca gtggggaata 360

ttggacaatg ggagaaatcc tgatccagca ataccgcgtg agtgatgaag gccttagggt 420

cgtaaaactc ttttgttagg aaagaaataa ttttaaataa tatttaaaat tgatgacggt 480

acctaaagaa taagcaccgg ctaactacgt gccagcagcc gcggtaatac gtagggtgca 540

agcgttaatc ggaattattg ggcgtaaaga gtgcgtaggc tgttatataa gatagatgtg 600

aaatacttaa gcttaactta agaactgcat ttattactgt ttaactagag tttattagag 660

agaagtggaa ttttatgtgt agcagtgaaa tgcgtagata tataaaggaa tatcgatggc 720

gaaggcagct tcttggaata atactgacgc tgaggcacga aagcgtgggg agcaaacagg 780

tagtattacc ctggtagtcc acgccctaaa ctatgtctac

aaatttagta acgtagctaa cgcattaagt agaccgcctg gggagtacga tcgcaagatt 900

aaaactcaaa ggaattgacg gggacccgca caagcggtgg atgatgtgga ttaattcgat 960

1020

gcttgaaaaa gaaccaaaac acaggtgctg catggctgtc gtcagctcgt gtcgtgagat 1080

gttgggttaa gtcccgcaac gagcgcaacc cttgttatta tttgctaata aaaagaactt 1140

taataagact gccaatgaca aattggagga aggtggggat gacgtcaagt cctcatggcc 1200

1260

1320

1380

tgtacacacc gcccgtcaca ccatggaagt gatttttacc agaaattatt tgtttaacct 1440

ttattggaaa aaaataatta aggtagaatt catgactggg gtgaagtcgt aacaaggtag 1500

cagtatcgga aggtgcggct ggattacatt ttaaat 1536

<210> 22

<211> 1423

<212> DNA

<213> Hodgkinia

<400> 22

aatgctggcg gcaggcctaa cacatgcaag tcgagcggac aacgttcaaa cgttgttagc 60

ggcgaacggg tgagtaatac gtgagaatct acccatccca acgtgataac atagtcaaca 120

180

cacgctagat tagctagttg gtgagataaa agcccaccaa ggccaagatc tatagctggt 240

ctggaaggat ggacagccac attgggactg agacaaggcc caaccctcta aggagggcag 300

360

aaggtccaac ggactgtaaa actcttttct ccagagatca taaatgatag tatctggtga 420

tataagctcc ggccaacttc gtgccagcag ccgcggtaat acgaggggag cgagtattgt 480

tcggttttat tgggcgtaaa gggtgtccag gttgctaagt aagttaacaa caaaatcttg 540

agattcaacc tcataacgtt cggttaatac tactaagctc gagcttggat agagacaaac 600

ggaattccga gtgtagaggt gaaattcgtt gatacttgga ggaacaccag aggcgaaggc 660

ggtttgtcat acccaagctga cactgaagac acgaaagcat ggggagcaaa caggattaga 720

taccctggta gtccatgcc taaacgttga gtgctaacag ttcgatcaag ccacatgcta 780

tgatccagga ttgtacagct aacgcgttaa gcactccgcc tgggtattac gaccgcaagg 840

ttaaaactca aaggaattga cggagacccg cacaagcggt ggagcatgtg gtttaattcg 900

aagctacacg aagaacctta ccagcccttg acataccatg gccaaccatc ctggaaacag 960

gatgttgttc aagttaaacc cttgaaatgc caggaacagg tgctgcatgg ctgttgtcag 1020

ttcgtgtcgt gagatgtatg gttaagtccc aaaacgaaca caaccctcac ccatagttgc 1080

cataaacaca attgggttct ctatgggtac tgctaacgta agttagagga aggtgaggac 1140

cacaacaagt catcatggcc cttatgggct gggccacaca catgctacaa tggtggttac 1200

aaagagccgc aacgttgtga gaccgagcaa atctccaaag accatctcag tccggattgt 1260

actctgcaac ccgagtacat gaagtaggaa tcgctagtaa tcgtggatca gcatgccacg 1320

1380

tcgaagtcaa gttacccttg accacatctt ggcaagtgac cga 1423

<210> 23

<211> 1504

<212> DNA

<213> Wolbachia sp. wPip

<400> 23

aaatttgaga gtttgatcct ggctcagaat gaacgctggc ggcaggccta acacatgcaa 60

gtcgaacgga gttatattgt agcttgctat ggtataactt agtggcagac gggtgagtaa 120

aattgttgga aacgacaact aataccgtat 180

acgccctacg ggggaaaaat ttattgctat tagatgagcc tatattagat tagctagttg 240

gtggggtaat agcctaccaa ggtaatgatc tatagctgat ctgagaggat gatcagccac 300

actggaactg agatacggtc cagactccta cgggaggcag cagtggggaa tattggacaa 360

tgggcgaaag cctgatccag ccatgccgca tgagtgaaga aggcctttgg gttgtaaagc 420

tcttttagtg aggaagataa tgacggtact cacagaagaa gtcctggcta actccgtgcc 480

agcagccgcg gtaatacgga gagggctagc gttattcgga attattgggc gtaaagggcg 540

cgtaggctgg ttaataagtt aaaagtgaaa tcccgaggct taaccttgga attgctttta 600

660

gtaaatatta ggaggaacac cagtggcgaa ggcgtctatc tggttcaaat ctgacgctga 720

agcgcgaagg cgtggggagc aaacaggatt agataccctg gtagtccacg ctgtaaacga 780

tgaatgttaa atatggggag tttactttct gtattacagc taacgcgtta aacattccgc 840

ctggggacta cggtcgcaag attaaaactc aaaggaattg acggggaccc gcacaagcgg 900

tggagcatgt ggtttaattc gatgcaacgc gaaaaacctt accacttctt gacatgaaaa 960

tcatacctat tcgaagggat agggtcggtt cggccggatt ttacacaagt gttgcatggc 1020

tgtcgtcagc tcgtgtcgtg agatgttggg ttaagtcccg caacgagcgc aaccctcatc 1080

cttagttgcc atcaggtaat gctgagtact ttaaggaaac tgccagtgat aagctggagg 1140

aaggtgggga tgatgtcaag tcatcatggc ctttatggag tgggctacac acgtgctaca 1200

atggtgtcta caatggggctg caaggtgcgc aagcctaagc taatccctaa aagacatctc 1260

agttcggatt gtactctgca actcgagtac atgaagttgg aatcgctagt aatcgtggat 1320

cagcatgcca cggtgaatac gttctcgggt cttgtacaca ctgcccgtca cgccatggga 1380

1440

agtgactggg gtgaagtcgt aacaaggtag cagtagggga atctgcagct ggattacctc 1500

ctta 1504

<210> 24

<211> 1532

<212> DNA

<213> Uzinura diaspidicola

<400> 24

aaaggagata ttccaaccac accttccggt acggttacct tgttacgact tagccctagt 60

catcaagttt accttaggca gaccactgaa ggattactga cttcaggtac ccccgactcc 120

catggcttga cgggcggtgt gtacaaggtt cgagaacata ttcaccgcgc cattgctgat 180

gcgcgattac tagcgattcc tgcttcatag agtcgaattg cagactccaa tccgaactga 240

gactggtttt agagattagc tcctgatcac ccagtggctg ccctttgtaa ccagccattg 300

tagcacgtgt gtagcccaag gcatagaggc catgatgatt tgacatcatc cccaccttcc 360

tcacagttta caccggcagt tttgttagag tccccggctt tacccgatgg caactaacaa 420

taggggttgc gctcgttata ggacttaacc aaacacttca cagcacgaac tgaagacaac 480

catgcagcac cttgtaatac gtcgtataga ctaagctgtt tccagcttat tcgtaataca 540

tttaagcctt ggtaaggttc ctcgcgtatc atcgaattaa accacatgct ccaccgcttg 600

tgcgaacccc cgtcaattcc tttgagtttc aatcttgcga ctgtacttcc caggtggatc 660

acttatcgct ttcgctaagc cactgaatat cgtttttcca atagctagtg atcatcgttt 720

agggcgtgga ctaccagggt atctaatcct gtttgctccc cacgctttcg tgcactgagc 780

gtcagtaaag atttagcaac ctgccttcgc tatcggtgtt ctgtatgata tctatgcatt 840

tcaccgctac accatacatt ccagatgctc caatcttact caagtttacc agtatcaata 900

gcaattttac agttaagctg taagctttca ctactgactt aataaacagc ctacacaccc 960

tttaaaccca ataaatccga ataacgcttg tgtcatccgt attgccgcgg ctgctggcac 1020

ggaattagcc gacacttatt cgtatagtac cttcaatctc ctatcacgta agatatttta 1080

tttctataca aaagcagttt acaacctaaa agaccttcat cctgcacgcg acgtagctgg 1140

ttcagagttt ccctccattga ccaatattcc tcactgctgc ctcccgtagg agtctggtcc 1200

gtgtctcagt accagtgtgg aggtacaccc tcttaggccc cctactgatc atagtcttgg 1260

tagagccatt acctcaccaa ctaactaatc aaacgcaggc tcatcttttg ccacctaagt 1320

tttaataaag gctccatgca gaaactttat attatggggg attaatcaga atttcttctg 1380

gctatacccc agcaaaaggt agattgcata cgtgttactc acccattcgc cggtcgccga 1440

caaattaaaa atttttcgat gcccctcgac ttgcatgtgt taagctcgcc gctagcgtta 1500

attctgagcc aggatcaaac tcttcgttgt ag 1532

<210> 25

<211> 1470

<212> DNA

<213> Carsonella ruddii

<400> 25

ctcaggataa acgctagcgg agggcttaac acatgcaagt cgaggggcag caaaaataat 60

tatttttggc gaccggcaaa cgggtgagta atacatacgt aactttcctt atgctgagga 120

atagcctgag gaaacttgga ttaatacctc ataatacaat tttttagaaa gaaaaattgt 180

taaagtttta ttatggcata agataggcgt atgtccaatt agttagttgg taaggtaatg 240

gcttaccaag acgatgattg gtagggggcc tgagaggggc gttcccccac attggtactg 300

agacacggac caaacttcta cggaaggctg cagtgaggaa tattggtcaa tggaggaaac 360

tctgaaccag ccactccgcg tgcaggatga aagaaagcct tattggttgt aaactgcttt 420

tgtatatgaa taaaaaattc taattataga aataattgaa ggtaatatac gaataagtat 480

cgactaactc tgtgccagca gtcgcggtaa gacagaggat acaagcgtta tccggattta 540

ttgggtttaa agggtgcgta ggcggttttt aaagtcagta gtgaaatctt aaagcttaac 600

660

tgtagcggtg aaatgcatag atatcacaca gaacaccgat agcgaaagca agttactaac 720

cctatactga cgctgagtca cgaaagcatg gggagcaaac aggattagat accctggtag 780

tccatgccgt aaacgatgat cactaactat tgggttttat acgttgtaat tcagtggtga 840

agcgaaagtg ttaagtgatc cacctgagga gtacgaccgc aaggttgaaa ctcaaaggaa 900

ttgacgggggg cccgcacaat cggtggagca tgtggtttaa ttcgatgata cacgaggaac 960

cttaccaaga cttaaatgta ctacgaataa attggaaaca atttagtcaa gcgacggagt 1020

1080

acgagcgcaa cccttattat tagttgccat cgagtaatgt caggggactc taataagact 1140

gccggcgcaa gccgagagga aggtggggat gacgtcaaat catcacggcc cttacgtctt 1200

gggccacaca cgtgctacaa tgatcggtac aaaagggagc gactgggtga ccaggagcaa 1260

atccagaaag ccgatctaag ttcggattgg agtctgaaac tcgactccat gaagctggaa 1320

1380

cgcccgtcaa gccatggaag ttggaagtac ctaaagttgg ttcgctacct aaggtaagtc 1440

taataactgg ggctaagtcg taacaaggta 1470

<210> 26

<211> 1761

<212> DNA

<213> Symbiotaphrina buchneri control strain JCM9740

<220>

<221> other_attribute

<222> (30)..(30)

<223> n is a, g, c or t

<220>

<221> other_attribute

<222> (40)..(40)

<223> n is a, g, c or t

<400> 26

agattaagcc atgcaagtct aagtataagn aatctatacn gtgaaactgc gaatggctca 60

ttaaatcagt tatcgtttat ttgatagtac cttactacat ggataaccgt ggtaattcta 120

gagctaatac atgctaaaaa ccccgacttc ggaaggggtg tatttattag ataaaaaacc 180

aatgcccttc ggggctcctt ggtgattcat gataacttaa cgaatcgcat ggccttgcgc 240

cggcgatggt tcattcaaat ttctgcccta tcaactttcg atggtaggat agtggcctac 300

catggtttta acgggtaacg gggaattagg gttcgattcc ggagagggag cctgagaaac 360

ggctaccaca tccaaggaag gcagcaggcg cgcaaattac ccaatcccga cacgggggagg 420

tagtgacaat aaatactgat acagggctct tttgggtctt gtaattggaa tgagtacaat 480

ttaaatccct taacgaggaa caattggagg gcaagtctgg tgccagcagc cgcggtaatt 540

ccagctccaa tagcgtatat taaagttgtt gcagttaaaa agctcgtagt tgaaccttgg 600

660

gagccgcatg cccttcactg ggtgtgtcgg ggaaccagga cttttacttt gaaaaaatta 720

gagtgttcaa agcaggccta tgctcgaata cattagcatg gaataataga ataggacgtg 780

cggttctatt ttgttggttt ctaggaccgc cgtaatgatt aatagggata gtcgggggca 840

tcagtattca attgtcagag gtgaaattct tggatttatt gaagactaac tactgcgaaa 900

gcatttgcca aggatgtttt cattaatcag tgaacgaaag ttaggggatc gaagacgatc 960

agataccgtc gtagtcttaa ccataaacta tgccgactag ggatcgggcg atgttattat 1020

tttgactcgc tcggcacctt acgagaaatc aaagtctttg ggttctgggg ggagtatggt 1080

cgcaaggctg aaacttaaag aaattgacgg aagggcacca ccaggagtgg agcctgcggc 1140

ttaatttgac tcaacacgggg gaaactcacc aggtccagac acattaagga ttgacagatt 1200

gagagctctt tcttgattat gtgggtggtg gtgcatggcc gttcttagtt ggtggagtga 1260

tttgtctgct taattgcgat aacgaacgag accttaacct gctaaaatagc ccggtccgct 1320

ttggcgggcc gctggcttct tagagggact atcggctcaa gccgatggaa gtttgaggca 1380

ataacaggtc tgtgatgccc ttagatgttc tgggccgcac gcgcgctaca ctgacagagc 1440

1500

gggatagagc attgcaatta ttgctcttca acgaggaatt cctagtaagc gcaagtcatc 1560

agcttgcgct gattacgtcc ctgccctttg tacacaccgc ccgtcgctac taccgattga 1620

atggctcagt gaggccttcg gactggcaca gggacgttgg caacgacgac ccagtgccgg 1680

1740

gaacctgcgg aggatcatt a 1761

<210> 27

<211> 1801

<212> DNA

<213> Symbiotaphrina kochii, control strain CBS 589.63

<220>

<221> other_attribute

<222> (1753)..(1755)

<223> n is a, g, c or t

<400> 27

tacctggttg attctgccag tagtcatatg cttgtctcaa agattaagcc atgcaagtct 60

aagtataagc aatctatacg gtgaaactgc gaatggctca ttaaatcagt tatcgtttat 120

ttgatagtac cttactacat ggataaccgt ggtaattcta gagctaatac atgctaaaaa 180

cctcgacttc ggaaggggtg tatttattag ataaaaaacc aatgcccttc ggggctcctt 240

ggtgattcat gataacttaa cgaatcgcat ggccttgcgc cggcgatggt tcattcaaat 300

ttctgcccta tcaactttcg atggtaggat agtggcctac catggtttca acgggtaacg 360

gggaattagg gttcgattcc ggagagggag cctgagaaac ggctaccaca tccaaggaag 420

gcagcaggcg cgcaaattac ccaatcccga cacgggggagg tagtgacaat aaatactgat 480

acagggctct tttggggtctt gtaattggaa tgagtacaat ttaaatccct taacgaggaa 540

caattggagg gcaagtctgg tgccagcagc cgcggtaatt ccagctccaa tagcgtatat 600

taaagttgtt gcagttaaaa agctcgtagt tgaaccttgg gcctggctgg ccggtccgcc 660

taaccgcgtg tactggtccg gccgggcctt tccttctggg gagccgcatg cccttcactg 720

ggtgtgtcgg ggaaccagga cttttacttt gaaaaaatta gagtgttcaa agcaggccta 780

tgctcgaata cattagcatg gaataataga ataggacgtg tggttctatt ttgttggttt 840

ctaggaccgc cgtaatgatt aatagggata gtcgggggca tcagtattca attgtcagag 900

gtgaaattct tggatttatt gaagactaac tactgcgaaa gcatttgcca aggatgtttt 960

cattaatcag tgaacgaaag ttaggggatc gaagacgatc agataccgtc gtagtcttaa 1020

1080

acgagaaatc aaagtctttg ggttctgggg ggagtatggt cgcaaggctg aaacttaaag 1140

aaattgacgg aagggcacca ccaggagtgg agcctgcggc ttaatttgac tcaacacggg 1200

gaaactcacc aggtccagac acattaagga ttgacagatt gagagctctt tcttgattat 1260

gtgggtggtg gtgcatggcc gttcttagtt ggtggagtga tttgtctgct taattgcgat 1320

aacgaacgag accttaacct gctaaaatagc ccggtccgct ttggcgggcc gctggcttct 1380

tagagggact atcggctcaa gccgatggaa gtttgaggca ataacaggtc tgtgatgccc 1440

ttagatgttc tgggccgcac gcgcgctaca ctgacagagc caacgagtac atcaccttgg 1500

ccggaaggtc tgggtaatct tgttaaactc tgtcgtgctg gggatagagc attgcaatta 1560

ttgctcttca acgaggaatt cctagtaagc gcaagtcatc agcttgcgct gattacgtcc 1620

ctgccctttg tacacaccgc ccgtcgctac taccgattga atggctcagt gaggccttcg 1680

gactggcaca gggacgttgg caacgacgac ccagtgccgg aaagttcgtc aaacttggtc 1740

aggatcatt 1800

a 1801

<210> 28

<211> 1490

<212> DNA

<213> Burkholderia sp. SFA1

<400> 28

agtttgatcc tggctcagat tgaacgctgg cggcatgcct tacacatgca agtcgaacgg 60

cagcacgggg gcaaccctgg tggcgagtgg cgaacgggtg agtaatacat cggaacgtgt 120

cctgtagtgg gggatagccc ggcgaaagcc ggattaatac cgcatacgac ctaagggaga 180

aagcggggga tcttcggacc tcgcgctata ggggcggccg atggcagatt agctagttgg 240

tggggtaaag gcctaccaag gcgacgatct gtagctggtc tgagaggacg accagccaca 300

ctgggactga gacacggccc agactcctac gggaggcagc agtggggaat tttggacaat 360

gggggcaacc ctgatccagc aatgccgcgt gtgtgaagaa ggcttcgggt tgtaaagcac 420

ttttgtccgg aaagaaaact tcgtccctaa tatggatgga ggatgacggt accggaagaa 480

taagcaccgg ctaactacgt gccagcagcc gcggtaatac gtagggtgcg agcgttaatc 540

ggaattactg ggcgtaaagc gtgcgcaggc ggtctgttaa gaccgatgtg aaatccccgg 600

gcttaacctg ggaactgcat tggtgactgg caggctttga gtgtggcaga ggggggtaga 660

attccacgtg tagcagtgaa atgcgtagag atgtggagga ataccgatgg cgaaggcagc 720

cccctgggcc aactactgac gctcatgcac gaaagcgtgg ggagcaaaca ggattagata 780

ccctggtagt ccacgcccta aacgatgtca actagttgtt ggggattcat ttccttagta 840

acgtagctaa cgcgtgaagt tgaccgcctg gggagtacgg tcgcaagatt aaaactcaaa 900

ggaattgacg gggacccgca caagcggtgg atgatgtgga ttaattcgat gcaacgcgaa 960

aaaccttacc tacccttgac atggtcggaa ccctgctgaa aggtgggggt gctcgaaaga 1020

gaaccggcgc acaggtgctg catggctgtc gtcagctcgt gtcgtgagat gttgggttaa 1080

gtcccgcaac gagcgcaacc cttgtcctta gttgctacgc aagagcactc taagggagact 1140

gccggtgaca aaccggagga aggtggggat gacgtcaagt cctcatggcc cttatgggta 1200

gggcttcaca cgtcatacaa tggtcggaac agaggggttgc caagccgcga ggtggagcca 1260

atcccagaaa accgatcgta gtccggatcg cagtctgcaa ctcgactgcg tgaagctgga 1320

atcgctagta atcgcggatc agcatgccgc ggtgaatacg ttcccgggtc ttgtacacac 1380

1440

cgcttaccac ggtgggattc atgactgggg tgaagtcgta acaaggtagc 1490

<210> 29

<211> 1408

<212> DNA

<213> Burkholderia sp. KM-A

<400> 29

gcaaccctgg tggcgagtgg cgaacgggtg agtaatacat cggaacgtgt cctgtagtgg 60

gggatagccc ggcgaaagcc ggattaatac cgcatacgat ctacggaaga aagcggggga 120

180

aggcctacca aggcgacgat ctgtagctgg tctgagagga cgaccagcca cactgggact 240

300

ccctgatcca gcaatgccgc gtgtgtgaag aaggccttcg ggttgtaaag cacttttgtc 360

cggaaagaaa acgtcttggt taatacctga ggcggatgac ggtaccggaa gaataagcac 420

cggctaacta cgtgccagca gccgcggtaa tacgtagggt gcgagcgtta atcggaatta 480

ctgggcgtaa agcgtgcgca ggcggtctgt taagaccgat gtgaaatccc cgggcttaac 540

ctgggaactg cattggtgac tggcaggctt tgagtgtggc agaggggggt agaattccac 600

gtgtagcagt gaaatgcgta gagatgtgga ggaataccga tggcgaaggc agccccctgg 660

gccaacactg acgctcatgc acgaaagcgt ggggagcaaa caggattaga taccctggta 720

780

aacgcgtgaa gttgaccgcc tggggagtac ggtcgcaaga ttaaaactca aaggaattga 840

cggggacccg cacaagcggt ggatgatgtg gattaattcg atgcaacgcg aaaaacctta 900

cctacccttg acatggtcgg aagtctgctg agaggtggac gtgctcgaaa gagaaccggc 960

gcacaggtgc tgcatggctg tcgtcagctc gtgtcgtgag atgttgggtt aagtcccgca 1020

acgagcgcaa cccttgtcct tagttgctac gcaagagcac tctaaggaga ctgccggtga 1080

caaaccggag gaaggtgggg atgacgtcaa gtcctcatgg cccttatggg tagggcttca 1140

cacgtcatac aatggtcgga acagagggtt gccaagccgc gaggtggagc caatcccaga 1200

aaaccgatcg tagtccggat cgcagtctgc aactcgactg cgtgaagctg gaatcgctag 1260

taatcgcgga tcagcatgcc gcggtgaata cgttcccggg tcttgtacac accgcccgtc 1320

acaccatggg agtgggtttc accagaagta ggtagcctaa ccgcaagggag ggcgcttacc 1380

acggtgggat tcatgactgg ggtgaagt 1408

<210> 30

<211> 1383

<212> DNA

<213> Burkholderia sp. KM-G

<400> 30

gcaaccctgg tggcgagtgg cgaacgggtg agtaatacat cggaacgtgt cctgtagtgg 60

gggatagccc ggcgaaagcc ggattaatac cgcatacgac ctaagggaga aagcggggga 120

tcttcggacc tcgcgctata ggggcggccg atggcagatt agctagttgg tggggtaaag 180

gcctaccaag gcgacgatct gtagctggtc tgagaggacg accagccaca ctgggactga 240

gacacggccc agactcctac gggaggcagc agtggggaat tttggacaat gggggcaacc 300

ctgatccagc aatgccgcgt gtgtgaagaa ggccttcggg ttgtaaagca cttttgtccg 360

gaaagaaaac ttcgaggtta atacccttgg aggatgacgg taccggaaga ataagcaccg 420

gctaactacg tgccagcagc cgcggtaata cgtagggtgc gagcgttaat cggaattact 480

gggcgtaaag cgtgcgcagg cggtctgtta agaccgatgt gaaatccccg ggcttaacct 540

gggaactgca ttggtgactg gcaggctttg agtgtggcag aggggggtag aattccacgt 600

gtagcagtga aatgcgtaga gatgtggagg aataccgatg gcgaaggcag ccccctgggc 660

caacactgac gctcatgcac gaaagcgtgg ggagcaaaca ggattagata ccctggtagt 720

ccacgcccta aacgatgtca actagttgtt ggggattcat ttccttagta acgtagctaa 780

cgcgtgaagt tgaccgcctg gggagtacgg tcgcaagatt aaaactcaaa ggaattgacg 840

gggacccgca caagcggtgg atgatgtgga ttaattcgat gcaacgcgaa aaaccttacc 900

tacccttgac atggtcggaa gtctgctgag aggtggacgt gctcgaaaga gaaccggcgc 960

acaggtgctg catggctgtc gtcagctcgt gtcgtgagat gttgggttaa gtcccgcaac 1020

gagcgcaacc cttgtcctta gttgctacgc aagagcactc taaggagact gccggtgaca 1080

aaccggagga aggtggggat gacgtcaagt cctcatggcc cttatgggta gggcttcaca 1140

1200

accgatcgta gtccggatcg cagtctgcaa ctcgactgcg tgaagctgga atcgctagta 1260

atcgcggatc agcatgccgc ggtgaatacg ttcccgggtc ttgtacacac cgcccgtcac 1320

accatgggag tgggtttcac cagaagtagg tagcctaacc tgcaaaggag ggcgcttacc 1380

acg 1383

<210> 31

<400> 31

000

<210> 32

<400> 32

000

<210> 33

<211> 1505

<212> DNA

<213> Snodgrassella alvi

<400> 33

gagagtttga tcctggctca gattgaacgc tggcggcatg ccttacacat gcaagtcgaa 60

cggcagcacg gagagcttgc tctctggtgg cgagtggcga acgggtgagt aatgcatcgg 120

aacgtaccga gtaatggggg ataactgtcc gaaaggatgg ctaataccgc atacgccctg 180

agggggaaag cgggggatcg aaagacctcg cgttatttga gcggccgatg ttggattagc 240

tagttggtgg ggtaaaggcc taccaaggcg acgatccata gcgggtctga gaggatgatc 300

360

ggacaatggg gggaaccctg atccagccat gccgcgtgtc tgaagaaggc cttcgggttg 420

taaaggactt ttgttaggga agaaaagccg ggtgttaata ccatctggtg ctgacggtac 480

ctaaagaata agcaccggct aactacgtgc cagcagccgc ggtaatacgt agggtgcgag 540

cgttaatcgg aattactggg cgtaaagcga gcgcagacgg ttaattaagt cagatgtgaa 600

atccccgagc tcaacttggg acgtgcattt gaaactggtt aactagagtg tgtcagaggg 660

aggtagaatt ccacgtgtag cagtgaaatg cgtagagatg tggaggaata ccgatggcga 720

aggcagcctc ctgggataac actgacgttc atgctcgaaa gcgtggggtag caaacaggat 780

tagataccct ggtagtccac gccctaaacg atgacaatta gctgttggga cactagatgt 840

cttagtagcg aagctaacgc gtgaaattgt ccgcctgggg agtacggtcg caagattaaa 900

actcaaagga attgacgggg acccgcacaa gcggtggatg atgtggatta attcgatgca 960

acgcgaagaa ccttacctgg tcttgacatg tacggaatct cttagagata ggagagtgcc 1020

ttcgggaacc gtaacacagg tgctgcatgg ctgtcgtcag ctcgtgtcgt gagatgttgg 1080

gttaagtccc gcaacgagcg caacccttgt cattagttgc catcattaag ttgggcactc 1140

taatgagact gccggtgaca aaccggagga aggtggggat gacgtcaagt cctcatggcc 1200

cttatgacca gggcttcaca cgtcatacaa tggtcggtac aggggtagc gaagccgcga 1260

ggtgaagcca atctcagaaa gccgatcgta gtccggattg cactctgcaa ctcgagtgca 1320

1380

ttgtacacac cgcccgtcac accatgggag tgggggatac cagaattgggg tagactaacc 1440

gcaaggaggt cgcttaacac ggtatgcttc atgactgggg tgaagtcgta acaaggtagc 1500

cgtag 1505

<210> 34

<211> 1541

<212> DNA

<213> Gilliamella apicola

<400> 34

ttaaattgaa gagtttgatc atggctcaga ttgaacgctg gcggcaggct taacacatgc 60

aagtcgaacg gtaacatgag tgcttgcact tgatgacgag tggcggacgg gtgagtaaag 120

tatggggatc tgccgaatgg agggggacaa cagttggaaa cgactgctaa taccgcataa 180

agttgagaga ccaaagcatg ggaccttcgg gccatgcgcc atttgatgaa cccatatggg 240

attagctagt tggtagggta atggcttacc aaggcgacga tctctagctg gtctgagagg 300

atgaccagcc acactggaac tgagacacgg tccagactcc tacgggaggc agcagtgggg 360

aatattgcac aatgggggaa accctgatgc agccatgccg cgtgtatgaa gaaggccttc 420

gggttgtaaa gtactttcgg tgatgaggaa ggtggtgtat ctaataggtg catcaattga 480

cgttaattac agaagaagca ccggctaact ccgtgccagc agccgcggta atacggaggg 540

tgcgagcgtt aatcggaatg actgggcgta aagggcatgt aggcggataa ttaagttagg 600

tgtgaaagcc ctgggctcaa cctaggaatt gcacttaaaa ctggttaact agagtattgt 660

agaggaaggt agaattccac gtgtagcggt gaaatgcgta gagatgtgga ggaataccgg 720

tggcgaaggc ggccttctgg acagatactg acgctgagat gcgaaagcgt ggggagcaaa 780

caggattaga taccctggta gtccacgctg taaacgatgt cgatttggag tttgttgcct 840

agagtgatgg gctccgaagc taacgcgata aatcgaccgc ctggggagta cggccgcaag 900

gttaaaactc aaatgaattg acggggggccc gcacaagcgg tggagcatgt ggtttaattc 960

gatgcaacgc gaagaacctt acctggtctt gacatccaca gaatcttgca gagatgcggg 1020

agtgccttcg ggaactgtga gacaggtgct gcatggctgt cgtcagctcg tgttgtgaaa 1080

1140

ggaactcaaa ggagactgcc gttgataaag cggaggaagg tggggacgac gtcaagtcat 1200

catggccctt acgaccaggg ctacacacgt gctacaatgg cgtatacaaa gggaggcgac 1260

ctcgcgagag caagcggacc tcataaagta cgtctaagtc cggattggag tctgcaactc 1320

gactccatga agtcggaatc gctagtaatc gtgaatcaga atgtcacggt gaatacgttc 1380

ccgggccttg tacacaccgc ccgtcacacc atgggagtgg gttgcaccag aagtagatag 1440

cttaaccttc gggagggcgt ttaccacggt gtggtccatg actggggtga agtcgtaaca 1500

aggtaaccgt aggggaacct gcggttggat cacctcctta c 1541

<210> 35

<211> 1528

<212> DNA

<213> Bartonella apis

<400> 35

aagccaaaat caaattttca acttgagagt ttgatcctgg ctcagaacga acgctggcgg 60

caggcttaac acatgcaagt cgaacgcact tttcggagtg agtggcagac gggtgagtaa 120

cgcgtgggaa tctacctatt tctacggaat aacgcagaga aatttgtgct aataccgtat 180

acgtccttcg ggagaaagat ttatcggaga tagatgagcc cgcgttggat tagctagttg 240

gtgaggtaat ggcccaccaa ggcgacgatc catagctggt ctgaggat gaccagccac 300

360

tgggcgcaag cctgatccag ccatgccgcg tgagtgatga aggccctagg gttgtaaagc 420

tctttcaccg gtgaagataa tgacggtaac cggagaagaa gccccggcta acttcgtgcc 480

agcagccgcg gtaatacgaa gggggctagc gttgttcgga tttactgggc gtaaagcgca 540

cgtaggcggga tatttaagtc aggggtgaaa tcccggggct caaccccggga actgcctttg 600

atactggata tcttgagtat ggaagaggta agtggaattc cgagtgtaga ggtgaaattc 660

gtagatattc ggaggaacac cagtggcgaa ggcggcttac tggtccatta ctgacgctga 720

ggtgcgaaag cgtggggagc aaacaggatt agataccctg gtagtccacg ctgtaaacga 780

tgaatgttag ccgttggaca gtttactgtt cggtggcgca gctaacgcat taaacattcc 840

gcctggggag tacggtcgca agattaaaac tcaaaggaat tgacgggggc ccgcacaagc 900

ggtggagcat gtggtttaat tcgaagcaac gcgcagaacc ttaccagccc ttgacatccc 960

gatcgcggat ggtggagaca ccgtctttca gttcggctgg atcggtgaca ggtgctgcat 1020

ggctgtcgtc agctcgtgtc gtgagatgtt gggttaagtc ccgcaacgag cgcaaccctc 1080

gcccttagtt gccatcattt agttgggcac tctaagggga ctgccggtga taagccgaga 1140

ggaaggtggg gatgacgtca agtcctcatg gcccttacgg gctgggctac acacgtgcta 1200

caatggtggt gacagtgggc agcgagaccg cgaggtcgag ctaatctcca aaagccatct 1260

cagttcggat tgcactctgc aactcgagtg catgaagttg gaatcgctag taatcgtgga 1320

1380

agttggtttt acccgaaggt gctgtgctaa ccgcaaggag gcaggcaacc acggtagggt 1440

cagcgactgg ggtgaagtcg taacaaggta gccgtagggg aacctgcggc tggatcacct 1500

cctttctaag gaagatgaag aattggaa 1528

<210> 36

<211> 1390

<212> DNA

<213> Parasaccharibacter apium

<220>

<221> other_attribute

<222> (643)..(756)

<223> n is a, g, c or t

<400> 36

ctaccatgca agtcgcacga aacctttcgg ggttagtggc ggacgggtga gtaacgcgtt 60

aggaacctat ctggaggtgg gggataacat cgggaaactg gtgctaatac cgcatgatgc 120

ctgaggggcca aggagagat ccgccattgg aggggcctgc gttcgattag ctagttggtt 180

gggtaaaggc tgaccaaggc gatgatcgat agctggtttg agaggatgat cagccacact 240

gggactgaga cacggcccag actcctacgg gaggcagcag tggggaatat tggacaatgg 300

gggcaaccct gatccagcaa tgccgcgtgt gtgaagaagg tcttcggatt gtaaagcact 360

ttcactaggg aagatgatga cggtacctag agaagaagcc ccggctaact tcgtgccagc 420

agccgcggta atacgaaggg ggctagcgtt gctcggaatg actgggcgta aagggcgcgt 480

aggctgtttg tacagtcaga tgtgaaatcc ccgggcttaa cctgggaact gcatttgata 540

cgtgcagact agagtccgag agagggttgt ggaattccca gtgtagaggt gaaattcgta 600

gatattggga agaacaccgg ttgcgaaggc ggcaacctgg ctnnnnnnnn nnnnnnnnnn 660

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 720

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnngagc taacgcgtta agcacaccgc 780

ctggggagta cggccgcaag gttgaaactc aaaggaattg acggggggccc gcacaagcgg 840

tggagcatgt ggtttaattc gaagcaacgc gcagaacctt accagggctt gcatggggag 900

gctgtattca gagatggata tttcttcgga cctcccgcac aggtgctgca tggctgtcgt 960

cagctcgtgt cgtgagatgt tgggttaagt cccgcaacga gcgcaaccct tgtctttagt 1020

tgccatcacg tctgggtggg cactctagag agactgccgg tgacaagccg gaggaaggtg 1080

gggatgacgt caagtcctca tggcccttat gtcctgggct acacacgtgc tacaatggcg 1140

gtgacagagg gatgctacat ggtgacatgg tgctgatctc aaaaaaccgt ctcagttcgg 1200

attgtactct gcaactcgag tgcatgaagg tggaatcgct agtaatcgcg gatcagcatg 1260

ccgcggtgaa tacgttcccg ggccttgtac acaccgcccg tcacaccatg ggagttggtt 1320

1380

gactggggga 1390

<210> 37

<211> 1583

<212> DNA

<213> Lactobacillus kunkeei

<400> 37

ttccttagaa aggaggtgat ccagccgcag gttctcctac ggctaccttg ttacgacttc 60

accctaatca tctgtcccac cttagacgac tagctcctaa aaggttaccc catcgtcttt 120

gggtgttaca aactctcatg gtgtgacggg cggtgtgtac aaggcccggg aacgtattca 180

ccgtggcatg ctgatccacg attactagtg attccaactt catgcaggcg agttgcagcc 240

tgcaatccga actgagaatg gctttaagag attagcttga cctcgcggtt tcgcgactcg 300

ttgtaccatc cattgtagca cgtgtgtagc ccagctcata aggggcatga tgatttgacg 360

tcgtccccac cttcctccgg tttatcaccg gcagtctcac tagagtgccc aactaaatgc 420

tggcaactaa taataagggt tgcgctcgtt gcgggactta acccaacatc tcacgacacg 480

agctgacgac aaccatgcac cacctgtcat tctgtccccg aagggaacgc ccaatctctt 540

gggttggcag aagatgtcaa gagctggtaa ggttcttcgc gtagcatcga attaaaccac 600

atgctccacc acttgtgcgg gcccccgtca attcctttga gtttcaacct tgcggtcgta 660

ctccccaggc ggaatactta atgcgttagc tgcggcactg aagggcggaa accctccaac 720

acctagtatt catcgtttac ggcatggact accagggtat ctaatcctgt tcgctaccca 780

tgctttcgag cctcagcgtc agtaacagac cagaaagccg ccttcgccac tggtgttctt 840

ccatatatct acgcatttca ccgctacaca tggagttcca ctttcctctt ctgtactcaa 900

gttttgtagt ttccactgca cttcctcagt tgagctgagg gctttcacag cagacttaca 960

aaaccgcctg cgctcgcttt acgcccaata aatccggaca acgcttgcca cctacgtatt 1020

accgcggctg ctggcacgta gttagccgtg gctttctggt taaataccgt caaagtgtta 1080

acagttactc taacacttgt tcttctttaa caacagagtt ttacgatccg aaaaccttca 1140

tcactcacgc ggcgttgctc catcagactt tcgtccattg tggaagattc cctactgctg 1200

cctcccgtag gagtctgggc cgtgtctcag tcccaatgtg gccgattacc ctctcaggtc 1260

ggctacgtat catcgtcttg gtgggctttt atctcaccaa ctaactaata cggcgcgggt 1320

ccatcccaaa gtgatagcaa agccatcttt caagttggaa ccatgcggtt ccaactaatt 1380

atgcggtatt agcacttgtt tccaaatgtt atcccccgct tcggggcagg ttacccacgt 1440

gttactcacc agttcgccac tcgctccgaa tccaaaaatc atttatgcaa gcataaaatc 1500

aatttgggag aactcgttcg acttgcatgt attaggcacg ccgccagcgt tcgtcctgag 1560

ccaggatcaa actctcatct taa 1583

<210> 38

<211> 1395

<212> DNA

<213> Lactobacillus Firm-4

<400> 38

acgaacgctg gcggcgtgcc taatacatgc aagtcgagcg cgggaagtca gggaagcctt 60

cgggtggaac tggtggaacg agcggcggat gggtgagtaa cacgtaggta acctgcccta 120

aagcggggga taccatctgg aaacaggtgc taataccgca taaacccagc agtcacatga 180

gtgctggttg aaagacggct tcggctgtca ctttaggatg gacctgcggc gtattagcta 240

gttggtggag taacggttca ccaaggcaat gatacgtagc cgacctgaga gggtaatcgg 300

ccacattggg actgagacac ggcccaaact cctacgggag gcagcagtag ggaatcttcc 360

acaatggacg caagtctgat ggagcaacgc cgcgtggatg aagaaggtct tcggatcgta 420

aaatcctgtt gttgaagaag aacggttgtg agagtaactg ctcataacgt gacggtaatc 480

aaccagaaag tcacggctaa ctacgtgcca gcagccgcgg taatacgtag gtggcaagcg 540

600

gccctcagct taactgagga agagcatcgg aaactgagag acttgagtgc agaagaggag 660

agtggaactc catgtgtagc ggtgaaatgc gtagatatat ggaagaacac cagtggcgaa 720

ggcggctctc tggtctgtta ctgacgctga ggctcgaaag catgggtagc gaacaggatt 780

agataccctg gtagtccatg ccgtaaacga tgagtgctaa gtgttgggag gtttccgcct 840

ctcagtgctg cagctaacgc attaagcact ccgcctgggg agtacgaccg caaggttgaa 900

actcaaagga attgacgggg gccgcacaa gcggtggagc atgtggttta attcgaagca 960

acgcgaagaa ccttaccagg tcttgacatc tcctgcaagc ctaagagatt aggggttccc 1020

ttcggggaca ggaagacagg tggtgcatgg ttgtcgtcag ctcgtgtcgt gagatgttgg 1080

1140

tagtgagact gccggtgaca aaccggagga aggtggggac gacgtcaaat catcatgccc 1200

cttatgacct gggctacaca cgtgctacaa tggatggtac aatgagaagc gaactcgcga 1260

ggggaagctg atctctgaaa accattctca gttcggattg caggctgcaa ctcgcctgca 1320

tgaagctgga atcgctagta atcgcggatc agcatgccgc ggtgaatacg ttcccggggcc 1380

ttgtacacac cgcc 1395

<210> 39

<211> 1549

<212> DNA

<213> Enterococcus

<400> 39

aggtgatcca gccgcacctt ccgatacggc taccttgtta cgacttcacc ccaatcatct 60

atcccacctt aggcggctgg ctccaaaaag gttacctcac cgacttcggg tgttacaaac 120

tctcgtggtg tgacgggcgg tgtgtacaag gcccgggaac gtattcaccg cggcgtgctg 180

atccgcgatt actagcgatt ccggcttcat gcaggcgagt tgcagcctgc aatccgaact 240

gagagaagct ttaagagatt tgcatgacct cgcggtctag cgactcgttg tacttcccat 300

tgtagcacgt gtgtagccca ggtcataagg ggcatgatga tttgacgtca tccccacctt 360

cctccggttt gtcaccggca gtctcgctag agtgcccaac taaatgatgg caactaacaa 420

taagggttgc gctcgttgcg ggacttaacc caacatctca cgacacgagc tgacgacaac 480

catgcaccac ctgtcacttt gtccccgaag ggaaagctct atctctagag tggtcaaagg 540

atgtcaagac ctggtaaggt tcttcgcgtt gcttcgaatt aaaccacatg ctccaccgct 600

tgtgcgggcc cccgtcaatt cctttgagtt tcaaccttgc ggtcgtactc cccaggcgga 660

gtgcttaatg cgtttgctgc agcactgaag ggcggaaacc ctccaacact tagcactcat 720

cgtttacggc gtggactacc agggtatcta atcctgtttg ctccccgc tttcgagcct 780

cagcgtcagt tacagaccag agagccgcct tcgccactgg tgttcctcca tatatctacg 840

catttcaccg ctacacatgg aattccactc tcctcttctg cactcaagtc tcccagtttc 900

caatgaccct ccccggttga gccgggggct ttcacatcag acttaagaaa ccgcctgcgc 960

tcgctttacg cccaataaat ccggacaacg cttgccacct acgtattacc gcggctgctg 1020

gcacgtagtt agccgtggct ttctggttag ataccgtcag gggacgttca gttactaacg 1080

tccttgttct tctctaacaa cagagtttta cgatccgaaa accttcttca ctcacgcggc 1140

gttgctcggt cagactttcg tccattgccg aagattccct actgctgcct cccgtaggag 1200

tctgggccgt gtctcagtcc cagtgtggcc gatcaccctc tcaggtcggc tatgcatcgt 1260

ggccttggtg agccgttacc tcaccaacta gctaatgcac cgcgggtcca tccatcagcg 1320

acacccgaaa gcgcctttca ctcttatgcc atgcggcata aactgttatg cggtattagc 1380

acctgtttcc aagtgttatc cccctctgat gggtaggtta cccacgtgtt actcacccgt 1440

ccgccactcc tctttccaat tgagtgcaag cactcgggag gaaagaagcg ttcgacttgc 1500

atgtattagg cacgccgcca gcgttcgtcc tgagccagga tcaaactct 1549

<210> 40

<211> 1541

<212> DNA

<213> Delftia

<400> 40

cagaaaggag gtgatccagc cgcaccttcc gatacggcta ccttgttacg acttcacccc 60

agtcacgaac cccgccgtgg taagcgccct ccttgcggtt aggctaccta cttctggcga 120

gacccgctcc catggtgtga cgggcggtgt gtacaagacc cgggaacgta ttcaccgcgg 180

catgctgatc cgcgattact agcgattccg acttcacgca gtcgagttgc agactgcgat 240

ccggactacg actggtttta tgggattagc tccccctcgc gggttggcaa ccctctgtac 300

cagccattgt atgacgtgtg tagccccacc tataagggcc atgaggactt gacgtcatcc 360

ccaccttcct ccggtttgtc accggcagtc tcattagagt gctcaactga atgtagcaac 420

taatgacaag ggttgcgctc gttgcgggac ttaacccaac atctcacgac acgagctgac 480

gacagccatg cagcacctgt gtgcaggttc tctttcgagc acgaatccat ctctggaaac 540

ttcctgccat gtcaaaggtg ggtaaggttt ttcgcgttgc atcgaattaa accacatcat 600

ccaccgcttg tgcgggtccc cgtcaattcc tttgagtttc aaccttgcgg ccgtactccc 660

caggcggtca acttcacgcg ttagcttcgt tactgagaaa actaattccc aacaaccagt 720

tgacatcgtt tagggcgtgg actaccaggg tatctaatcc tgtttgctcc ccacgctttc 780

gtgcatgagc gtcagtacag gtccagggga ttgccttcgc catcggtgtt cctccgcata 840

tctacgcatt tcactgctac acgcggaatt ccatccccct ctaccgtact ctagccatgc 900

agtcacaaat gcagttccca ggttgagccc ggggatttca catctgtctt acataaccgc 960

ctgcgcacgc tttacgccca gtaattccga ttaacgctcg caccctacgt attaccgcgg 1020

ctgctggcac gtagttagcc ggtgcttatt cttacggtac cgtcatgggc cccctgtatt 1080

agaaggagct ttttcgttcc gtacaaaagc agtttacaac ccgaaggcct tcatcctgca 1140

cgcggcattg ctggatcagg ctttcgccca ttgtccaaaa ttccccactg ctgcctcccg 1200

taggagtctg ggccgtgtct cagtcccagt gtggctggtc gtcctctcag accagctaca 1260

gatcgtcggc ttggtaagct tttatcccac caactaccta atctgccatc ggccgctcca 1320

atcgcgcgag gcccgaaggg cccccgcttt catcctcaga tcgtatgcgg tattagctac 1380

tctttcgagt agttatcccc cacgactggg cacgttccga tgtattactc acccgttcgc 1440

cactcgtcag cgtccgaaga cctgttaccg ttcgacttgc atgtgtaagg catgccgcca 1500

gcgttcaatc tgagccagga tcaaactcta cagttcgatc t 1541

<210> 41

<211> 1502

<212> DNA

<213> Pelomonas

<220>

<221> other_attribute

<222> (192)..(193)

<223> n is a, g, c or t

<220>

<221> other_attribute

<222> (832)..(833)

<223> n is a, g, c or t

<400> 41

atcctggctc agattgaacg ctggcggcat gccttacaca tgcaagtcga acggtaacag 60

gttaagctga cgagtggcga acgggtgagt aatatatcgg aacgtgccca gtcgtggggg 120

ataactgctc gaaagagcag ctaataccgc atacgacctg agggtgaaag cgggggatcg 180

caagacctcg cnngattgga gcggccgata tcagattagg tagttggtgg ggtaaaggcc 240

caccaagcca acgatctgta gctggtctga gaggacgacc agccacactg ggactgagac 300

acggcccaga ctcctacggg aggcagcagt ggggaatttt ggacaatgggg cgcaagcctg 360

atccagccat gccgcgtgcg ggaagaaggc cttcgggttg taaaccgctt ttgtcaggga 420

agaaaaggtt ctggttaata cctgggactc atgacggtac ctgaagaata agcaccggct 480

aactacgtgc cagcagccgc ggtaatacgt agggtgcaag cgttaatcgg aattactggg 540

cgtaaagcgt gcgcaggcgg ttatgcaaga cagaggtgaa atcccggggc tcaacctggg 600

aactgccttt gtgactgcat agctagagta cggtagaggg ggatggaatt ccgcgtgtag 660

cagtgaaatg cgtagatatg cggaggaaca ccgatggcga aggcaatccc ctggacctgt 720

actgacgctc atgcacgaaa gcgtggggag caaacaggat tagataccct ggtagtccac 780

gccctaaacg atgtcaactg gttgttggga gggtttcttc tcagtaacgt anntaacgcg 840

tgaagttgac cgcctgggga gtacggccgc aaggttgaaa ctcaaaggaa ttgacgggga 900

cccgcacaag cggtggatga tgtggtttaa ttcgatgcaa cgcgaaaaac cttacctacc 960

cttgacatgc caggaatcct gaagagattt gggagtgctc gaaagagaac ctggacacag 1020

gtgctgcatg gccgtcgtca gctcgtgtcg tgagatgttg ggttaagtcc cgcaacgagc 1080

gcaacccttg tcattagttg ctacgaaagg gcactctaat gagactgccg gtgacaaacc 1140

ggaggaaggt ggggatgacg tcaggtcatc atggccctta tgggtagggc tacacacgtc 1200

atacaatggc cgggacagag ggctgccaac ccgcgagggg gagctaatcc cagaaacccg 1260

gtcgtagtcc ggatcgtagt ctgcaactcg actgcgtgaa gtcggaatcg ctagtaatcg 1320

cggatcagct tgccgcggtg aatacgttcc cgggtcttgt acacaccgcc cgtcacacca 1380

tgggagcggg ttctgccaga agtagttagc ctaaccgcaa ggagggcgat taccacggca 1440

gggttcgtga ctggggtgaa gtcgtaacaa ggtagccgta tcggaaggtg cggctggatc 1500

ac 1502

<210> 42

<211> 34

<212> PROTEIN

<213> Lactococcus lactis

<400> 42

Ile Thr Ser Ile Ser Leu Cys Thr Pro Gly Cys Lys Thr Gly Ala Leu

1 5 10 15

Met Gly Cys Asn Met Lys Thr Ala Thr Cys His Cys Ser Ile His Val

20 25 30

Ser Lys

<210> 43

<211> 22

<212> PROTEIN

<213> Staphylococcus epidermis

<400> 43

Ile Ala Ser Lys Phe Ile Cys Thr Pro Gly Cys Ala Lys Thr Gly Ser

1 5 10 15

Phe Asn Ser Tyr Cys Cys

twenty

<210> 44

<211> 44

<212> PROTEIN

<213> Pediococcus acidilactici

<400> 44

Lys Tyr Tyr Gly Asn Gly Val Thr Cys Gly Lys His Ser Cys Ser Val

1 5 10 15

Asp Trp Gly Lys Ala Thr Thr Cys Ile Ile Asn Asn Gly Ala Met Ala

20 25 30

Trp Ala Thr Gly Gly His Gln Gly Asn His Lys Cys

35 40

<210> 45

<211> 44

<212> PROTEIN

<213> Enterococcus faecium

<400> 45

Ala Thr Arg Ser Tyr Gly Asn Gly Val Tyr Cys Asn Asn Ser Lys Cys

1 5 10 15

Trp Val Asn Trp Gly Glu Ala Lys Glu Asn Ile Ala Gly Ile Val Ile

20 25 30

Ser Gly Trp Ala Ser Gly Leu Ala Gly Met Gly His

35 40

<210> 46

<211> 39

<212> PROTEIN

<213> Streptococcus lactis

<400> 46

Gly Thr Trp Asp Asp Ile Gly Gln Gly Ile Gly Arg Val Ala Tyr Trp

1 5 10 15

Val Gly Lys Ala Met Gly Asn Met Ser Asp Val Asn Gln Ala Ser Arg

20 25 30

Ile Asn Arg Lys Lys Lys His

35

<210> 47

<211> 48

<212> PROTEIN

<213> Lactobacillus johnsonii

<400> 47

Asn Arg Trp Gly Asp Thr Val Leu Ser Ala Ala Ser Gly Ala Gly Thr

1 5 10 15

Gly Ile Lys Ala Cys Lys Ser Phe Gly Pro Trp Gly Met Ala Ile Cys

20 25 30

Gly Val Gly Gly Ala Ala Ile Gly Gly Tyr Phe Gly Tyr Thr His Asn

35 40 45

<210> 48

<211> 70

<212> PROTEIN

<213> Enterococcus faecalis

<400> 48

Met Ala Lys Glu Phe Gly Ile Pro Ala Ala Val Ala Gly Thr Val Leu

1 5 10 15

Asn Val Val Glu Ala Gly Gly Trp Val Thr Thr Ile Val Ser Ile Leu

20 25 30

Thr Ala Val Gly Ser Gly Gly Leu Ser Leu Leu Ala Ala Ala Gly Arg

35 40 45

Glu Ser Ile Lys Ala Tyr Leu Lys Lys Glu Ile Lys Lys Lys Gly Lys

50 55 60

Arg Ala Val Ile Ala Trp

65 70

<210> 49

<211> 51

<212> PROTEIN

<213> Staphylococcus aureus

<400> 49

Met Ser Trp Leu Asn Phe Leu Lys Tyr Ile Ala Lys Tyr Gly Lys Lys

1 5 10 15

Ala Val Ser Ala Ala Trp Lys Tyr Lys Gly Lys Val Leu Glu Trp Leu

20 25 30

Asn Val Gly Pro Thr Leu Glu Trp Val Trp Gln Lys Leu Lys Lys Ile

35 40 45

Ala Gly Leu

fifty

<210> 50

<211> 43

<212> PROTEIN

<213> Lactococcus garvieae

<400> 50

Ile Gly Gly Ala Leu Gly Asn Ala Leu Asn Gly Leu Gly Thr Trp Ala

1 5 10 15

Asn Met Met Asn Gly Gly Gly Phe Val Asn Gln Trp Gln Val Tyr Ala

20 25 30

Asn Lys Gly Lys Ile Asn Gln Tyr Arg Pro Tyr

35 40

<210> 51

<211> 103

<212> PROTEIN

<213> Escherichia coli

<400> 51

Met Arg Thr Leu Thr Leu Asn Glu Leu Asp Ser Val Ser Gly Gly Ala

1 5 10 15

Ser Gly Arg Asp Ile Ala Met Ala Ile Gly Thr Leu Ser Gly Gln Phe

20 25 30

Val Ala Gly Gly Ile Gly Ala Ala Ala Gly Gly Val Ala Gly Gly Ala

35 40 45

Ile Tyr Asp Tyr Ala Ser Thr His Lys Pro Asn Pro Ala Met Ser Pro

50 55 60

Ser Gly Leu Gly Gly Thr Ile Lys Gln Lys Pro Glu Gly Ile Pro Ser

65 70 75 80

Glu Ala Trp Asn Tyr Ala Ala Gly Arg Leu Cys Asn Trp Ser Pro Asn

85 90 95

Asn Leu Ser Asp Val Cys Leu

100

<210> 52

<211> 339

<212> PROTEIN

<213> Cp1

<400> 52

Met Val Lys Lys Asn Asp Leu Phe Val Asp Val Ser Ser His Asn Gly

1 5 10 15

Tyr Asp Ile Thr Gly Ile Leu Glu Gln Met Gly Thr Thr Asn Thr Ile

20 25 30

Ile Lys Ile Ser Glu Ser Thr Thr Tyr Leu Asn Pro Cys Leu Ser Ala

35 40 45

Gln Val Glu Gln Ser Asn Pro Ile Gly Phe Tyr His Phe Ala Arg Phe

50 55 60

Gly Gly Asp Val Ala Glu Ala Glu Arg Glu Ala Gln Phe Phe Leu Asp

65 70 75 80

Asn Val Pro Met Gln Val Lys Tyr Leu Val Leu Asp Tyr Glu Asp Asp

85 90 95

Pro Ser Gly Asp Ala Gln Ala Asn Thr Asn Ala Cys Leu Arg Phe Met

100 105 110

Gln Met Ile Ala Asp Ala Gly Tyr Lys Pro Ile Tyr Tyr Ser Tyr Lys

115 120 125

Pro Phe Thr His Asp Asn Val Asp Tyr Gln Gln Ile Leu Ala Gln Phe

130 135 140

Pro Asn Ser Leu Trp Ile Ala Gly Tyr Gly Leu Asn Asp Gly Thr Ala

145 150 155 160

Asn Phe Glu Tyr Phe Pro Ser Met Asp Gly Ile Arg Trp Trp Gln Tyr

165 170 175

Ser Ser Asn Pro Phe Asp Lys Asn Ile Val Leu Leu Asp Asp Glu Glu

180 185 190

Asp Asp Lys Pro Lys Thr Ala Gly Thr Trp Lys Gln Asp Ser Lys Gly

195 200 205

Trp Trp Phe Arg Arg Asn Asn Gly Ser Phe Pro Tyr Asn Lys Trp Glu

210 215 220

Lys Ile Gly Gly Val Trp Tyr Tyr Phe Asp Ser Lys Gly Tyr Cys Leu

225 230 235 240

Thr Ser Glu Trp Leu Lys Asp Asn Glu Lys Trp Tyr Tyr Leu Lys Asp

245 250 255

Asn Gly Ala Met Ala Thr Gly Trp Val Leu Val Gly Ser Glu Trp Tyr

260 265 270

Tyr Met Asp Asp Ser Gly Ala Met Val Thr Gly Trp Val Lys Tyr Lys

275 280 285

Asn Asn Trp Tyr Tyr Met Thr Asn Glu Arg Gly Asn Met Val Ser Asn

290 295 300

Glu Phe Ile Lys Ser Gly Lys Gly Trp Tyr Phe Met Asn Thr Asn Gly

305 310 315 320

Glu Leu Ala Asp Asn Pro Ser Phe Thr Lys Glu Pro Asp Gly Leu Ile

325 330 335

Thr Val Ala

<210> 53

<211> 296

<212> PROTEIN

<213> Dp-1

<400> 53

Met Gly Val Asp Ile Glu Lys Gly Val Ala Trp Met Gln Ala Arg Lys

1 5 10 15

Gly Arg Val Ser Tyr Ser Met Asp Phe Arg Asp Gly Pro Asp Ser Tyr

20 25 30

Asp Cys Ser Ser Ser Met Tyr Tyr Ala Leu Arg Ser Ala Gly Ala Ser

35 40 45

Ser Ala Gly Trp Ala Val Asn Thr Glu Tyr Met His Ala Trp Leu Ile

50 55 60

Glu Asn Gly Tyr Glu Leu Ile Ser Glu Asn Ala Pro Trp Asp Ala Lys

65 70 75 80

Arg Gly Asp Ile Phe Ile Trp Gly Arg Lys Gly Ala Ser Ala Gly Ala

85 90 95

Gly Gly His Thr Gly Met Phe Ile Asp Ser Asp Asn Ile Ile His Cys

100 105 110

Asn Tyr Ala Tyr Asp Gly Ile Ser Val Asn Asp His Asp Glu Arg Trp

115 120 125

Tyr Tyr Ala Gly Gln Pro Tyr Tyr Tyr Val Tyr Arg Leu Thr Asn Ala

130 135 140

Asn Ala Gln Pro Ala Glu Lys Lys Leu Gly Trp Gln Lys Asp Ala Thr

145 150 155 160

Gly Phe Trp Tyr Ala Arg Ala Asn Gly Thr Tyr Pro Lys Asp Glu Phe

165 170 175

Glu Tyr Ile Glu Glu Asn Lys Ser Trp Phe Tyr Phe Asp Asp Gln Gly

180 185 190

Tyr Met Leu Ala Glu Lys Trp Leu Lys His Thr Asp Gly Asn Trp Tyr

195 200 205

Trp Phe Asp Arg Asp Gly Tyr Met Ala Thr Ser Trp Lys Arg Ile Gly

210 215 220

Glu Ser Trp Tyr Tyr Phe Asn Arg Asp Gly Ser Met Val Thr Gly Trp

225 230 235 240

Ile Lys Tyr Tyr Asp Asn Trp Tyr Tyr Cys Asp Ala Thr Asn Gly Asp

245 250 255

Met Lys Ser Asn Ala Phe Ile Arg Tyr Asn Asp Gly Trp Tyr Leu Leu

260 265 270

Leu Pro Asp Gly Arg Leu Ala Asp Lys Pro Gln Phe Thr Val Glu Pro

275 280 285

Asp Gly Leu Ile Thr Ala Lys Val

290 295

<210> 54

<211> 233

<212> PROTEIN

<213> gamma

<400> 54

Met Glu Ile Gln Lys Lys Leu Val Asp Pro Ser Lys Tyr Gly Thr Lys

1 5 10 15

Cys Pro Tyr Thr Met Lys Pro Lys Tyr Ile Thr Val His Asn Thr Tyr

20 25 30

Asn Asp Ala Pro Ala Glu Asn Glu Val Ser Tyr Met Ile Ser Asn Asn

35 40 45

Asn Glu Val Ser Phe His Ile Ala Val Asp Asp Lys Lys Ala Ile Gln

50 55 60

Gly Ile Pro Leu Glu Arg Asn Ala Trp Ala Cys Gly Asp Gly Asn Gly

65 70 75 80

Ser Gly Asn Arg Gln Ser Ile Ser Val Glu Ile Cys Tyr Ser Lys Ser

85 90 95

Gly Gly Asp Arg Tyr Tyr Lys Ala Glu Asp Asn Ala Val Asp Val Val

100 105 110

Arg Gln Leu Met Ser Met Tyr Asn Ile Pro Ile Glu Asn Val Arg Thr

115 120 125

His Gln Ser Trp Ser Gly Lys Tyr Cys Pro His Arg Met Leu Ala Glu

130 135 140

Gly Arg Trp Gly Ala Phe Ile Gln Lys Val Lys Asn Gly Asn Val Ala

145 150 155 160

Thr Thr Ser Pro Thr Lys Gln Asn Ile Ile Gln Ser Gly Ala Phe Ser

165 170 175

Pro Tyr Glu Thr Pro Asp Val Met Gly Ala Leu Thr Ser Leu Lys Met

180 185 190

Thr Ala Asp Phe Ile Leu Gln Ser Asp Gly Leu Thr Tyr Phe Ile Ser

195 200 205

Lys Pro Thr Ser Asp Ala Gln Leu Lys Ala Met Lys Glu Tyr Leu Asp

210 215 220

Arg Lys Gly Trp Trp Tyr Glu Val Lys

225 230

<210> 55

<211> 481

<212> PROTEIN

<213> MR11

<400> 55

Met Gln Ala Lys Leu Thr Lys Lys Glu Phe Ile Glu Trp Leu Lys Thr

1 5 10 15

Ser Glu Gly Lys Gln Phe Asn Val Asp Leu Trp Tyr Gly Phe Gln Cys

20 25 30

Phe Asp Tyr Ala Asn Ala Gly Trp Lys Val Leu Phe Gly Leu Leu Leu

35 40 45

Lys Gly Leu Gly Ala Lys Asp Ile Pro Phe Ala Asn Asn Phe Asp Gly

50 55 60

Leu Ala Thr Val Tyr Gln Asn Thr Pro Asp Phe Leu Ala Gln Pro Gly

65 70 75 80

Asp Met Val Val Phe Gly Ser Asn Tyr Gly Ala Gly Tyr Gly His Val

85 90 95

Ala Trp Val Ile Glu Ala Thr Leu Asp Tyr Ile Ile Val Tyr Glu Gln

100 105 110

Asn Trp Leu Gly Gly Gly Trp Thr Asp Arg Ile Glu Gln Pro Gly Trp

115 120 125

Gly Trp Glu Lys Val Thr Arg Arg Gln His Ala Tyr Asp Phe Pro Met

130 135 140

Trp Phe Ile Arg Pro Asn Phe Lys Ser Glu Thr Ala Pro Arg Ser Ile

145 150 155 160

Gln Ser Pro Thr Gln Ala Ser Lys Lys Glu Thr Ala Lys Pro Gln Pro

165 170 175

Lys Ala Val Glu Leu Lys Ile Ile Lys Asp Val Val Lys Gly Tyr Asp

180 185 190

Leu Pro Lys Arg Gly Gly Asn Pro Lys Gly Ile Val Ile His Asn Asp

195 200 205

Ala Gly Ser Lys Gly Ala Thr Ala Glu Ala Tyr Arg Asn Gly Leu Val

210 215 220

Asn Ala Pro Leu Ser Arg Leu Glu Ala Gly Ile Ala His Ser Tyr Val

225 230 235 240

Ser Gly Asn Thr Val Trp Gln Ala Leu Asp Glu Ser Gln Val Gly Trp

245 250 255

His Thr Ala Asn Gln Leu Gly Asn Lys Tyr Tyr Tyr Gly Ile Glu Val

260 265 270

Cys Gln Ser Met Gly Ala Asp Asn Ala Thr Phe Leu Lys Asn Glu Gln

275 280 285

Ala Thr Phe Gln Glu Cys Ala Arg Leu Leu Lys Lys Trp Gly Leu Pro

290 295 300

Ala Asn Arg Asn Thr Ile Arg Leu His Asn Glu Phe Thr Ser Thr Ser

305 310 315 320

Cys Pro His Arg Ser Ser Val Leu His Thr Gly Phe Asp Pro Val Thr

325 330 335

Arg Gly Leu Leu Pro Glu Asp Lys Gln Leu Gln Leu Lys Asp Tyr Phe

340 345 350

Ile Lys Gln Ile Arg Val Tyr Met Asp Gly Lys Ile Pro Val Ala Thr

355 360 365

Val Ser Asn Glu Ser Ser Ala Ser Ser Asn Thr Val Lys Pro Val Ala

370 375 380

Ser Ala Trp Lys Arg Asn Lys Tyr Gly Thr Tyr Tyr Met Glu Glu Asn

385 390 395 400

Ala Arg Phe Thr Asn Gly Asn Gln Pro Ile Thr Val Arg Lys Ile Gly

405 410 415

Pro Phe Leu Ser Cys Pro Val Ala Tyr Gln Phe Gln Pro Gly Gly Tyr

420 425 430

Cys Asp Tyr Thr Glu Val Met Leu Gln Asp Gly His Val Trp Val Gly

435 440 445

Tyr Thr Trp Glu Gly Gln Arg Tyr Tyr Leu Pro Ile Arg Thr Trp Asn

450 455 460

Gly Ser Ala Pro Pro Asn Gln Ile Leu Gly Asp Leu Trp Gly Glu Ile

465 470 475 480

Ser

<210> 56

<211> 239

<212> PROTEIN

<213> B30

<400> 56

Met Val Ile Asn Ile Glu Gln Ala Ile Ala Trp Met Ala Ser Arg Lys

1 5 10 15

Gly Lys Val Thr Tyr Ser Met Asp Tyr Arg Asn Gly Pro Ser Ser Tyr

20 25 30

Asp Cys Ser Ser Ser Val Tyr Phe Ala Leu Arg Ser Ala Gly Ala Ser

35 40 45

Asp Asn Gly Trp Ala Val Asn Thr Glu Tyr Glu His Asp Trp Leu Ile

50 55 60

Lys Asn Gly Tyr Val Leu Ile Ala Glu Asn Thr Asn Trp Asn Ala Gln

65 70 75 80

Arg Gly Asp Ile Phe Ile Trp Gly Lys Arg Gly Ala Ser Ala Gly Ala

85 90 95

Phe Gly His Thr Gly Met Phe Val Asp Pro Asp Asn Ile Ile His Cys

100 105 110

Asn Tyr Gly Tyr Asn Ser Ile Thr Val Asn Asn His Asp Glu Ile Trp

115 120 125

Gly Tyr Asn Gly Gln Pro Tyr Val Tyr Ala Tyr Arg Tyr Ser Gly Lys

130 135 140

Gln Ser Asn Ala Lys Val Asp Asn Lys Ser Val Val Ser Lys Phe Glu

145 150 155 160

Lys Glu Leu Asp Val Asn Thr Pro Leu Ser Asn Ser Asn Met Pro Tyr

165 170 175

Tyr Glu Ala Thr Ile Ser Glu Asp Tyr Tyr Val Glu Ser Lys Pro Asp

180 185 190

Val Asn Ser Thr Asp Lys Glu Leu Leu Val Ala Gly Thr Arg Val Arg

195 200 205

Val Tyr Glu Lys Val Lys Gly Trp Ala Arg Ile Gly Ala Pro Gln Ser

210 215 220

Asn Gln Trp Val Glu Asp Ala Tyr Leu Ile Asp Ala Thr Asp Met

225 230 235

<210> 57

<211> 495

<212> PROTEIN

<213> K

<400> 57

Met Ala Lys Thr Gln Ala Glu Ile Asn Lys Arg Leu Asp Ala Tyr Ala

1 5 10 15

Lys Gly Thr Val Asp Ser Pro Tyr Arg Val Lys Lys Ala Thr Ser Tyr

20 25 30

Asp Pro Ser Phe Gly Val Met Glu Ala Gly Ala Ile Asp Ala Asp Gly

35 40 45

Tyr Tyr His Ala Gln Cys Gln Asp Leu Ile Thr Asp Tyr Val Leu Trp

50 55 60

Leu Thr Asp Asn Lys Val Arg Thr Trp Gly Asn Ala Lys Asp Gln Ile

65 70 75 80

Lys Gln Ser Tyr Gly Thr Gly Phe Lys Ile His Glu Asn Lys Pro Ser

85 90 95

Thr Val Pro Lys Lys Gly Trp Ile Ala Val Phe Thr Ser Gly Ser Tyr

100 105 110

Glu Gln Trp Gly His Ile Gly Ile Val Tyr Asp Gly Gly Asn Thr Ser

115 120 125

Thr Phe Thr Ile Leu Glu Gln Asn Trp Asn Gly Tyr Ala Asn Lys Lys

130 135 140

Pro Thr Lys Arg Val Asp Asn Tyr Tyr Gly Leu Thr His Phe Ile Glu

145 150 155 160

Ile Pro Val Lys Ala Gly Thr Thr Val Lys Lys Glu Thr Ala Lys Lys

165 170 175

Ser Ala Ser Lys Thr Pro Ala Pro Lys Lys Lys Ala Thr Leu Lys Val

180 185 190

Ser Lys Asn His Ile Asn Tyr Thr Met Asp Lys Arg Gly Lys Lys Pro

195 200 205

Glu Gly Met Val Ile His Asn Asp Ala Gly Arg Ser Ser Gly Gln Gln

210 215 220

Tyr Glu Asn Ser Leu Ala Asn Ala Gly Tyr Ala Arg Tyr Ala Asn Gly

225 230 235 240

Ile Ala His Tyr Tyr Gly Ser Glu Gly Tyr Val Trp Glu Ala Ile Asp

245 250 255

Ala Lys Asn Gln Ile Ala Trp His Thr Gly Asp Gly Thr Gly Ala Asn

260 265 270

Ser Gly Asn Phe Arg Phe Ala Gly Ile Glu Val Cys Gln Ser Met Ser

275 280 285

Ala Ser Asp Ala Gln Phe Leu Lys Asn Glu Gln Ala Val Phe Gln Phe

290 295 300

Thr Ala Glu Lys Phe Lys Glu Trp Gly Leu Thr Pro Asn Arg Lys Thr

305 310 315 320

Val Arg Leu His Met Glu Phe Val Pro Thr Ala Cys Pro His Arg Ser

325 330 335

Met Val Leu His Thr Gly Phe Asn Pro Val Thr Gln Gly Arg Pro Ser

340 345 350

Gln Ala Ile Met Asn Lys Leu Lys Asp Tyr Phe Ile Lys Gln Ile Lys

355 360 365

Asn Tyr Met Asp Lys Gly Thr Ser Ser Ser Thr Val Val Lys Asp Gly

370 375 380

Lys Thr Ser Ser Ala Ser Thr Pro Ala Thr Arg Pro Val Thr Gly Ser

385 390 395 400

Trp Lys Lys Asn Gln Tyr Gly Thr Trp Tyr Lys Pro Glu Asn Ala Thr

405 410 415

Phe Val Asn Gly Asn Gln Pro Ile Val Thr Arg Ile Gly Ser Pro Phe

420 425 430

Leu Asn Ala Pro Val Gly Gly Asn Leu Pro Ala Gly Ala Thr Ile Val

435 440 445

Tyr Asp Glu Val Cys Ile Gln Ala Gly His Ile Trp Ile Gly Tyr Asn

450 455 460

Ala Tyr Asn Gly Asn Arg Val Tyr Cys Pro Val Arg Thr Cys Gln Gly

465 470 475 480

Val Pro Pro Asn Gln Ile Pro Gly Val Ala Trp Gly Val Phe Lys

485 490 495

<210> 58

<211> 281

<212> PROTEIN

<213>A118

<400> 58

Met Thr Ser Tyr Tyr Tyr Ser Arg Ser Leu Ala Asn Val Asn Lys Leu

1 5 10 15

Ala Asp Asn Thr Lys Ala Ala Ala Arg Lys Leu Leu Asp Trp Ser Glu

20 25 30

Ser Asn Gly Ile Glu Val Leu Ile Tyr Glu Thr Ile Arg Thr Lys Glu

35 40 45

Gln Gln Ala Ala Asn Val Asn Ser Gly Ala Ser Gln Thr Met Arg Ser

50 55 60

Tyr His Leu Val Gly Gln Ala Leu Asp Phe Val Met Ala Lys Gly Lys

65 70 75 80

Thr Val Asp Trp Gly Ala Tyr Arg Ser Asp Lys Gly Lys Lys Phe Val

85 90 95

Ala Lys Ala Lys Ser Leu Gly Phe Glu Trp Gly Gly Asp Trp Ser Gly

100 105 110

Phe Val Asp Asn Pro His Leu Gln Phe Asn Tyr Lys Gly Tyr Gly Thr

115 120 125

Asp Thr Phe Gly Lys Gly Ala Ser Thr Ser Asn Ser Ser Lys Pro Ser

130 135 140

Ala Asp Thr Asn Thr Asn Ser Leu Gly Leu Val Asp Tyr Met Asn Leu

145 150 155 160

Asn Lys Leu Asp Ser Ser Phe Ala Asn Arg Lys Lys Leu Ala Thr Ser

165 170 175

Tyr Gly Ile Lys Asn Tyr Ser Gly Thr Ala Thr Gln Asn Thr Thr Leu

180 185 190

Leu Ala Lys Leu Lys Ala Gly Lys Pro His Thr Pro Ala Ser Lys Asn

195 200 205

Thr Tyr Tyr Thr Glu Asn Pro Arg Lys Val Lys Thr Leu Val Gln Cys

210 215 220

Asp Leu Tyr Lys Ser Val Asp Phe Thr Thr Lys Asn Gln Thr Gly Gly

225 230 235 240

Thr Phe Pro Pro Gly Thr Val Phe Thr Ile Ser Gly Met Gly Lys Thr

245 250 255

Lys Gly Gly Thr Pro Arg Leu Lys Thr Lys Ser Gly Tyr Tyr Leu Thr

260 265 270

Ala Asn Thr Lys Phe Val Lys Lys Ile

275 280

<210> 59

<211> 341

<212> PROTEIN

<213> A511

<400> 59

Met Val Lys Tyr Thr Val Glu Asn Lys Ile Ile Ala Gly Leu Pro Lys

1 5 10 15

Gly Lys Leu Lys Gly Ala Asn Phe Val Ile Ala His Glu Thr Ala Asn

20 25 30

Ser Lys Ser Thr Ile Asp Asn Glu Val Ser Tyr Met Thr Arg Asn Trp

35 40 45

Lys Asn Ala Phe Val Thr His Phe Val Gly Gly Gly Gly Arg Val Val

50 55 60

Gln Val Ala Asn Val Asn Tyr Val Ser Trp Gly Ala Gly Gln Tyr Ala

65 70 75 80

Asn Ser Tyr Ser Tyr Ala Gln Val Glu Leu Cys Arg Thr Ser Asn Ala

85 90 95

Thr Thr Phe Lys Lys Asp Tyr Glu Val Tyr Cys Gln Leu Leu Val Asp

100 105 110

Leu Ala Lys Lys Ala Gly Ile Pro Ile Thr Leu Asp Ser Gly Ser Lys

115 120 125

Thr Ser Asp Lys Gly Ile Lys Ser His Lys Trp Val Ala Asp Lys Leu

130 135 140

Gly Gly Thr Thr His Gln Asp Pro Tyr Ala Tyr Leu Ser Ser Trp Gly

145 150 155 160

Ile Ser Lys Ala Gln Phe Ala Ser Asp Leu Ala Lys Val Ser Gly Gly

165 170 175

Gly Asn Thr Gly Thr Ala Pro Ala Lys Pro Ser Thr Pro Ala Pro Lys

180 185 190

Pro Ser Thr Pro Ser Thr Asn Leu Asp Lys Leu Gly Leu Val Asp Tyr

195 200 205

Met Asn Ala Lys Lys Met Asp Ser Ser Tyr Ser Asn Arg Asp Lys Leu

210 215 220

Ala Lys Gln Tyr Gly Ile Ala Asn Tyr Ser Gly Thr Ala Ser Gln Asn

225 230 235 240

Thr Thr Leu Leu Ser Lys Ile Lys Gly Gly Ala Pro Lys Pro Ser Thr

245 250 255

Pro Ala Pro Lys Pro Ser Thr Ser Thr Ala Lys Lys Ile Tyr Phe Pro

260 265 270

Pro Asn Lys Gly Asn Trp Ser Val Tyr Pro Thr Asn Lys Ala Pro Val

275 280 285

Lys Ala Asn Ala Ile Gly Ala Ile Asn Pro Thr Lys Phe Gly Gly Leu

290 295 300

Thr Tyr Thr Ile Gln Lys Asp Arg Gly Asn Gly Val Tyr Glu Ile Gln

305 310 315 320

Thr Asp Gln Phe Gly Arg Val Gln Val Tyr Gly Ala Pro Ser Thr Gly

325 330 335

Ala Val Ile Lys Lys

340

<210> 60

<211> 289

<212> PROTEIN

<213> A500

<400> 60

Met Ala Leu Thr Glu Ala Trp Leu Ile Glu Lys Ala Asn Arg Lys Leu

1 5 10 15

Asn Ala Gly Gly Met Tyr Lys Ile Thr Ser Asp Lys Thr Arg Asn Val

20 25 30

Ile Lys Lys Met Ala Lys Glu Gly Ile Tyr Leu Cys Val Ala Gln Gly

35 40 45

Tyr Arg Ser Thr Ala Glu Gln Asn Ala Leu Tyr Ala Gln Gly Arg Thr

50 55 60

Lys Pro Gly Ala Ile Val Thr Asn Ala Lys Gly Gly Gln Ser Asn His

65 70 75 80

Asn Tyr Gly Val Ala Val Asp Leu Cys Leu Tyr Thr Asn Asp Gly Lys

85 90 95

Asp Val Ile Trp Glu Ser Thr Thr Ser Arg Trp Lys Lys Val Val Ala

100 105 110

Ala Met Lys Ala Glu Gly Phe Lys Trp Gly Gly Asp Trp Lys Ser Phe

115 120 125

Lys Asp Tyr Pro His Phe Glu Leu Cys Asp Ala Val Ser Gly Glu Lys

130 135 140

Ile Pro Ala Ala Thr Gln Asn Thr Asn Thr Asn Ser Asn Arg Tyr Glu

145 150 155 160

Gly Lys Val Ile Asp Ser Ala Pro Leu Leu Pro Lys Met Asp Phe Lys

165 170 175

Ser Ser Pro Phe Arg Met Tyr Lys Val Gly Thr Glu Phe Leu Val Tyr

180 185 190

Asp His Asn Gln Tyr Trp Tyr Lys Thr Tyr Ile Asp Asp Lys Leu Tyr

195 200 205

Tyr Met Tyr Lys Ser Phe Cys Asp Val Val Ala Lys Lys Asp Ala Lys

210 215 220

Gly Arg Ile Lys Val Arg Ile Lys Ser Ala Lys Asp Leu Arg Ile Pro

225 230 235 240

Val Trp Asn Asn Ile Lys Leu Asn Ser Gly Lys Ile Lys Trp Tyr Ala

245 250 255

Pro Asn Val Lys Leu Ala Trp Tyr Asn Tyr Arg Arg Gly Tyr Leu Glu

260 265 270

Leu Trp Tyr Pro Asn Asp Gly Trp Tyr Tyr Thr Ala Glu Tyr Phe Leu

275 280 285

Lys

<210> 61

<211> 239

<212> PROTEIN

<213> Prophage LambdaSa1

<400> 61

Met Val Ile Asn Ile Glu Gln Ala Ile Ala Trp Met Ala Ser Arg Lys

1 5 10 15

Gly Lys Val Thr Tyr Ser Met Asp Tyr Arg Asn Gly Pro Ser Ser Tyr

20 25 30

Asp Cys Ser Ser Ser Val Tyr Phe Ala Leu Arg Ser Ala Gly Ala Ser

35 40 45

Asp Asn Gly Trp Ala Val Asn Thr Glu Tyr Glu His Asp Trp Leu Ile

50 55 60

Lys Asn Gly Tyr Val Leu Ile Ala Glu Asn Thr Asn Trp Asn Ala Gln

65 70 75 80

Arg Gly Asp Ile Phe Ile Trp Gly Lys Arg Gly Ala Ser Ala Gly Ala

85 90 95

Phe Gly His Thr Gly Met Phe Val Asp Pro Asp Asn Ile Ile His Cys

100 105 110

Asn Tyr Gly Tyr Asn Ser Ile Thr Val Asn Asn His Asp Glu Ile Trp

115 120 125

Gly Tyr Asn Gly Gln Pro Tyr Val Tyr Ala Tyr Arg Tyr Ala Arg Lys

130 135 140

Gln Ser Asn Ala Lys Val Asp Asn Gln Ser Val Val Ser Lys Phe Glu

145 150 155 160

Lys Glu Leu Asp Val Asn Thr Pro Leu Ser Asn Ser Asn Met Pro Tyr

165 170 175

Tyr Glu Ala Thr Ile Ser Glu Asp Tyr Tyr Val Glu Ser Lys Pro Asp

180 185 190

Val Asn Ser Thr Asp Lys Glu Leu Leu Val Ala Gly Thr Arg Val Arg

195 200 205

Val Tyr Glu Lys Val Lys Gly Trp Ala Arg Ile Gly Ala Pro Gln Ser

210 215 220

Asn Gln Trp Val Glu Asp Ala Tyr Leu Ile Asp Ala Thr Asp Met

225 230 235

<210> 62

<211> 468

<212> PROTEIN

<213> Prophage LambdaSa2

<400> 62

Met Glu Ile Asn Thr Glu Ile Ala Ile Ala Trp Met Ser Ala Arg Gln

1 5 10 15

Gly Lys Val Ser Tyr Ser Met Asp Tyr Arg Asp Gly Pro Asn Ser Tyr

20 25 30

Asp Cys Ser Ser Ser Val Tyr Tyr Ala Leu Arg Ser Ala Gly Ala Ser

35 40 45

Ser Ala Gly Trp Ala Val Asn Thr Glu Tyr Met His Asp Trp Leu Ile

50 55 60

Lys Asn Gly Tyr Glu Leu Ile Ala Glu Asn Val Asp Trp Asn Ala Val

65 70 75 80

Arg Gly Asp Ile Ala Ile Trp Gly Met Arg Gly His Ser Ser Gly Ala

85 90 95

Gly Gly His Val Val Met Phe Ile Asp Pro Glu Asn Ile Ile His Cys

100 105 110

Asn Trp Ala Asn Asn Gly Ile Thr Val Asn Asn Tyr Asn Gln Thr Ala

115 120 125

Ala Ala Ser Gly Trp Met Tyr Cys Tyr Val Tyr Arg Leu Lys Ser Gly

130 135 140

Ala Ser Thr Gln Gly Lys Ser Leu Asp Thr Leu Val Lys Glu Thr Leu

145 150 155 160

Ala Gly Asn Tyr Gly Asn Gly Glu Ala Arg Lys Ala Val Leu Gly Asn

165 170 175

Gln Tyr Glu Ala Val Met Ser Val Ile Asn Gly Lys Thr Thr Thr Asn

180 185 190

Gln Lys Thr Val Asp Gln Leu Val Gln Glu Val Ile Ala Gly Lys His

195 200 205

Gly Asn Gly Glu Ala Arg Lys Lys Ser Leu Gly Ser Gln Tyr Asp Ala

210 215 220

Val Gln Lys Arg Val Thr Glu Leu Leu Lys Lys Gln Pro Ser Glu Pro

225 230 235 240

Phe Lys Ala Gln Glu Val Asn Lys Pro Thr Glu Thr Lys Thr Ser Gln

245 250 255

Thr Glu Leu Thr Gly Gln Ala Thr Ala Thr Lys Glu Glu Gly Asp Leu

260 265 270

Ser Phe Asn Gly Thr Ile Leu Lys Lys Ala Val Leu Asp Lys Ile Leu

275 280 285

Gly Asn Cys Lys Lys His Asp Ile Leu Pro Ser Tyr Ala Leu Thr Ile

290 295 300

Leu His Tyr Glu Gly Leu Trp Gly Thr Ser Ala Val Gly Lys Ala Asp

305 310 315 320

Asn Asn Trp Gly Gly Met Thr Trp Thr Gly Gln Gly Asn Arg Pro Ser

325 330 335

Gly Val Thr Val Thr Gln Gly Ser Ala Arg Pro Ser Asn Glu Gly Gly

340 345 350

His Tyr Met His Tyr Ala Ser Val Asp Asp Phe Leu Thr Asp Trp Phe

355 360 365

Tyr Leu Leu Arg Ala Gly Gly Ser Tyr Lys Val Ser Gly Ala Lys Thr

370 375 380

Phe Ser Glu Ala Ile Lys Gly Met Phe Lys Val Gly Gly Ala Val Tyr

385 390 395 400

Asp Tyr Ala Ala Ser Gly Phe Asp Ser Tyr Ile Val Gly Ala Ser Ser

405 410 415

Arg Leu Lys Ala Ile Glu Ala Glu Asn Gly Ser Leu Asp Lys Phe Asp

420 425 430

Lys Ala Thr Asp Ile Gly Asp Gly Ser Lys Asp Lys Ile Asp Ile Thr

435 440 445

Ile Glu Gly Ile Glu Val Thr Ile Asn Gly Ile Thr Tyr Glu Leu Thr

450 455 460

Lys Lys Pro Val

465

<210> 63

<211> 236

<212> PROTEIN

<213> Prophage (ATCC700407)

<400> 63

Met Thr Asp Ser Ile Gln Glu Met Arg Lys Leu Gln Ser Ile Pro Val

1 5 10 15

Arg Tyr Asp Met Gly Asp Arg Tyr Gly Asn Asp Ala Asp Arg Asp Gly

20 25 30

Arg Ile Glu Met Asp Cys Ser Ser Ala Val Ser Lys Ala Leu Gly Ile

35 40 45

Ser Met Thr Asn Asn Thr Glu Thr Leu Gln Gln Ala Leu Pro Ala Ile

50 55 60

Gly Tyr Gly Lys Ile His Asp Ala Val Asp Gly Thr Phe Asp Met Gln

65 70 75 80

Ala Tyr Asp Val Ile Ile Trp Ala Pro Arg Asp Gly Ser Ser Ser Leu

85 90 95

Gly Ala Phe Gly His Val Leu Ile Ala Thr Ser Pro Thr Thr Ala Ile

100 105 110

His Cys Asn Tyr Gly Ser Asp Gly Ile Thr Glu Asn Asp Tyr Asn Tyr

115 120 125

Ile Trp Glu Ile Asn Gly Arg Pro Arg Glu Ile Val Phe Arg Lys Gly

130 135 140

Val Thr Gln Thr Gln Ala Thr Val Thr Ser Gln Phe Glu Arg Glu Leu

145 150 155 160

Asp Val Asn Ala Arg Leu Thr Val Ser Asp Lys Pro Tyr Tyr Glu Ala

165 170 175

Thr Leu Ser Glu Asp Tyr Tyr Val Glu Ala Gly Pro Arg Ile Asp Ser

180 185 190

Gln Asp Lys Glu Leu Ile Lys Ala Gly Thr Arg Val Arg Val Tyr Glu

195 200 205

Lys Leu Asn Gly Trp Ser Arg Ile Asn His Pro Glu Ser Ala Gln Trp

210 215 220

Val Glu Asp Ser Tyr Leu Val Asp Ala Thr Glu Met

225 230 235

<210> 64

<211> 481

<212> PROTEIN

<213> Phi11 and Phi12

<400> 64

Met Gln Ala Lys Leu Thr Lys Asn Glu Phe Ile Glu Trp Leu Lys Thr

1 5 10 15

Ser Glu Gly Lys Gln Phe Asn Val Asp Leu Trp Tyr Gly Phe Gln Cys

20 25 30

Phe Asp Tyr Ala Asn Ala Gly Trp Lys Val Leu Phe Gly Leu Leu Leu

35 40 45

Lys Gly Leu Gly Ala Lys Asp Ile Pro Phe Ala Asn Asn Phe Asp Gly

50 55 60

Leu Ala Thr Val Tyr Gln Asn Thr Pro Asp Phe Leu Ala Gln Pro Gly

65 70 75 80

Asp Met Val Val Phe Gly Ser Asn Tyr Gly Ala Gly Tyr Gly His Val

85 90 95

Ala Trp Val Ile Glu Ala Thr Leu Asp Tyr Ile Ile Val Tyr Glu Gln

100 105 110

Asn Trp Leu Gly Gly Gly Trp Thr Asp Gly Ile Glu Gln Pro Gly Trp

115 120 125

Gly Trp Glu Lys Val Thr Arg Arg Gln His Ala Tyr Asp Phe Pro Met

130 135 140

Trp Phe Ile Arg Pro Asn Phe Lys Ser Glu Thr Ala Pro Arg Ser Val

145 150 155 160

Gln Ser Pro Thr Gln Ala Pro Lys Lys Glu Thr Ala Lys Pro Gln Pro

165 170 175

Lys Ala Val Glu Leu Lys Ile Ile Lys Asp Val Val Lys Gly Tyr Asp

180 185 190

Leu Pro Lys Arg Gly Ser Asn Pro Lys Gly Ile Val Ile His Asn Asp

195 200 205

Ala Gly Ser Lys Gly Ala Thr Ala Glu Ala Tyr Arg Asn Gly Leu Val

210 215 220

Asn Ala Pro Leu Ser Arg Leu Glu Ala Gly Ile Ala His Ser Tyr Val

225 230 235 240

Ser Gly Asn Thr Val Trp Gln Ala Leu Asp Glu Ser Gln Val Gly Trp

245 250 255

His Thr Ala Asn Gln Ile Gly Asn Lys Tyr Tyr Tyr Gly Ile Glu Val

260 265 270

Cys Gln Ser Met Gly Ala Asp Asn Ala Thr Phe Leu Lys Asn Glu Gln

275 280 285

Ala Thr Phe Gln Glu Cys Ala Arg Leu Leu Lys Lys Trp Gly Leu Pro

290 295 300

Ala Asn Arg Asn Thr Ile Arg Leu His Asn Glu Phe Thr Ser Thr Ser

305 310 315 320

Cys Pro His Arg Ser Ser Val Leu His Thr Gly Phe Asp Pro Val Thr

325 330 335

Arg Gly Leu Leu Pro Glu Asp Lys Arg Leu Gln Leu Lys Asp Tyr Phe

340 345 350

Ile Lys Gln Ile Arg Ala Tyr Met Asp Gly Lys Ile Pro Val Ala Thr

355 360 365

Val Ser Asn Glu Ser Ser Ala Ser Ser Asn Thr Val Lys Pro Val Ala

370 375 380

Ser Ala Trp Lys Arg Asn Lys Tyr Gly Thr Tyr Tyr Met Glu Glu Ser

385 390 395 400

Ala Arg Phe Thr Asn Gly Asn Gln Pro Ile Thr Val Arg Lys Val Gly

405 410 415

Pro Phe Leu Ser Cys Pro Val Gly Tyr Gln Phe Gln Pro Gly Gly Tyr

420 425 430

Cys Asp Tyr Thr Glu Val Met Leu Gln Asp Gly His Val Trp Val Gly

435 440 445

Tyr Thr Trp Glu Gly Gln Arg Tyr Tyr Leu Pro Ile Arg Thr Trp Asn

450 455 460

Gly Ser Ala Pro Pro Asn Gln Ile Leu Gly Asp Leu Trp Gly Glu Ile

465 470 475 480

Ser

<210> 65

<211> 481

<212> PROTEIN

<213> (Phi)H5

<400> 65

Met Gln Ala Lys Leu Thr Lys Lys Glu Phe Ile Glu Trp Leu Lys Thr

1 5 10 15

Ser Glu Gly Lys Gln Tyr Asn Ala Asp Gly Trp Tyr Gly Phe Gln Cys

20 25 30

Phe Asp Tyr Ala Asn Ala Gly Trp Lys Ala Leu Phe Gly Leu Leu Leu

35 40 45

Lys Gly Val Gly Ala Lys Asp Ile Pro Phe Ala Asn Asn Phe Asp Gly

50 55 60

Leu Ala Thr Val Tyr Gln Asn Thr Pro Asp Phe Leu Ala Gln Pro Gly

65 70 75 80

Asp Met Val Val Phe Gly Ser Asn Tyr Gly Ala Gly Tyr Gly His Val

85 90 95

Ala Trp Val Ile Glu Ala Thr Leu Asp Tyr Ile Ile Val Tyr Glu Gln

100 105 110

Asn Trp Leu Gly Gly Gly Trp Thr Asp Gly Val Gln Gln Pro Gly Ser

115 120 125

Gly Trp Glu Lys Val Thr Arg Arg Gln His Ala Tyr Asp Phe Pro Met

130 135 140

Trp Phe Ile Arg Pro Asn Phe Lys Ser Glu Thr Ala Pro Arg Ser Val

145 150 155 160

Gln Ser Pro Thr Gln Ala Ser Lys Lys Glu Thr Ala Lys Pro Gln Pro

165 170 175

Lys Ala Val Glu Leu Lys Ile Ile Lys Asp Val Val Lys Gly Tyr Asp

180 185 190

Leu Pro Lys Arg Gly Ser Asn Pro Asn Phe Ile Val Ile His Asn Asp

195 200 205

Ala Gly Ser Lys Gly Ala Thr Ala Glu Ala Tyr Arg Asn Gly Leu Val

210 215 220

Asn Ala Pro Leu Ser Arg Leu Glu Ala Gly Ile Ala His Ser Tyr Val

225 230 235 240

Ser Gly Asn Thr Val Trp Gln Ala Leu Asp Glu Ser Gln Val Gly Trp

245 250 255

His Thr Ala Asn Gln Ile Gly Asn Lys Tyr Gly Tyr Gly Ile Glu Val

260 265 270

Cys Gln Ser Met Gly Ala Asp Asn Ala Thr Phe Leu Lys Asn Glu Gln

275 280 285

Ala Thr Phe Gln Glu Cys Ala Arg Leu Leu Lys Lys Trp Gly Leu Pro

290 295 300

Ala Asn Arg Asn Thr Ile Arg Leu His Asn Glu Phe Thr Ser Thr Ser

305 310 315 320

Cys Pro His Arg Ser Ser Val Leu His Thr Gly Phe Asp Pro Val Thr

325 330 335

Arg Gly Leu Leu Pro Glu Asp Lys Arg Leu Gln Leu Lys Asp Tyr Phe

340 345 350

Ile Lys Gln Ile Arg Ala Tyr Met Asp Gly Lys Ile Pro Val Ala Thr

355 360 365

Val Ser Asn Asp Ser Ser Ala Ser Ser Asn Thr Val Lys Pro Val Ala

370 375 380

Ser Ala Trp Lys Arg Asn Lys Tyr Gly Thr Tyr Tyr Met Glu Glu Ser

385 390 395 400

Ala Arg Phe Thr Asn Gly Asn Gln Pro Ile Thr Val Arg Lys Val Gly

405 410 415

Pro Phe Leu Ser Cys Pro Val Gly Tyr Gln Phe Gln Pro Gly Gly Tyr

420 425 430

Cys Asp Tyr Thr Glu Val Met Leu Gln Asp Gly His Val Trp Val Gly

435 440 445

Tyr Thr Trp Glu Gly Gln Arg Tyr Tyr Leu Pro Ile Arg Thr Trp Asn

450 455 460

Gly Ser Ala Pro Pro Asn Gln Ile Leu Gly Asp Leu Trp Gly Glu Ile

465 470 475 480

Ser

<210> 66

<211> 477

<212> PROTEIN

<213> (Phi)WMY

<400> 66

Met Lys Thr Lys Ala Gln Ala Lys Ser Trp Ile Asn Ser Lys Ile Gly

1 5 10 15

Lys Gly Ile Asp Trp Asp Gly Met Tyr Gly Tyr Gln Cys Met Asp Glu

20 25 30

Ala Val Asp Tyr Ile His His Val Thr Asp Gly Lys Val Thr Met Trp

35 40 45

Gly Asn Ala Ile Asp Ala Pro Lys Asn Asn Phe Gln Gly Leu Cys Thr

50 55 60

Val Tyr Thr Asn Thr Pro Glu Phe Arg Pro Ala Tyr Gly Asp Val Ile

65 70 75 80

Val Trp Ser Tyr Gly Thr Phe Ala Thr Tyr Gly His Ile Ala Ile Val

85 90 95

Val Asn Pro Asp Pro Tyr Gly Asp Leu Gln Tyr Ile Thr Val Leu Glu

100 105 110

Gln Asn Trp Asn Gly Asn Gly Ile Tyr Lys Thr Glu Phe Ala Thr Ile

115 120 125

Arg Thr His Asp Tyr Thr Gly Val Ser His Phe Ile Arg Pro Lys Phe

130 135 140

Ala Asp Glu Val Lys Glu Thr Ala Lys Thr Val Asn Lys Leu Ser Val

145 150 155 160

Gln Lys Lys Ile Val Thr Pro Lys Asn Ser Val Glu Arg Ile Lys Asn

165 170 175

Tyr Val Lys Thr Ser Gly Tyr Ile Asn Gly Glu His Tyr Glu Leu Tyr

180 185 190

Asn Arg Gly His Lys Pro Lys Gly Val Val Ile His Asn Thr Ala Gly

195 200 205

Thr Ala Ser Ala Thr Gln Glu Gly Gln Arg Leu Thr Asn Met Thr Phe

210 215 220

Gln Gln Leu Ala Asn Gly Val Ala His Val Tyr Ile Asp Lys Asn Thr

225 230 235 240

Ile Tyr Glu Thr Leu Pro Glu Asp Arg Ile Ala Trp His Val Ala Gln

245 250 255

Gln Tyr Gly Asn Thr Glu Phe Tyr Gly Ile Glu Val Cys Gly Ser Arg

260 265 270

Asn Thr Asp Lys Glu Gln Phe Leu Ala Asn Glu Gln Val Ala Phe Gln

275 280 285

Glu Ala Ala Arg Arg Leu Lys Ser Trp Gly Met Arg Ala Asn Arg Asn

290 295 300

Thr Val Arg Leu His Thr Phe Ser Ser Thr Glu Cys Pro Asp Met

305 310 315 320

Ser Met Leu Leu His Thr Gly Tyr Ser Met Lys Asn Gly Lys Pro Thr

325 330 335

Gln Asp Ile Thr Asn Lys Cys Ala Asp Tyr Phe Met Lys Gln Ile Asn

340 345 350

Ala Tyr Ile Asp Gly Lys Gln Pro Thr Ser Thr Val Val Gly Ser Ser

355 360 365

Ser Ser Asn Lys Leu Lys Ala Lys Asn Lys Asp Lys Ser Thr Gly Trp

370 375 380

Asn Thr Asn Glu Tyr Gly Thr Leu Trp Lys Lys Glu His Ala Thr Phe

385 390 395 400

Thr Cys Gly Val Arg Gln Gly Ile Val Thr Arg Thr Thr Gly Pro Phe

405 410 415

Thr Ser Cys Pro Gln Ala Gly Val Leu Tyr Tyr Gly Gln Ser Val Asn

420 425 430

Tyr Asp Thr Val Cys Lys Gln Asp Gly Tyr Val Trp Ile Ser Trp Thr

435 440 445

Thr Ser Asp Gly Tyr Asp Val Trp Met Pro Ile Arg Thr Trp Asp Arg

450 455 460

Ser Thr Asp Lys Val Ser Glu Ile Trp Gly Thr Ile Ser

465 470 475

<210> 67

<211> 443

<212> PROTEIN

<213> (Phi)NCTC 11261

<400> 67

Met Ala Thr Tyr Gln Glu Tyr Lys Ser Arg Ser Asn Gly Asn Ala Tyr

1 5 10 15

Asp Ile Asp Gly Ser Phe Gly Ala Gln Cys Trp Asp Gly Tyr Ala Asp

20 25 30

Tyr Cys Lys Tyr Leu Gly Leu Pro Tyr Ala Asn Cys Thr Asn Thr Gly

35 40 45

Tyr Ala Arg Asp Ile Trp Glu Gln Arg His Glu Asn Gly Ile Leu Asn

50 55 60

Tyr Phe Asp Glu Val Glu Val Met Gln Ala Gly Asp Val Ala Ile Phe

65 70 75 80

Met Val Val Asp Gly Val Thr Pro Tyr Ser His Val Ala Ile Phe Asp

85 90 95

Ser Asp Ala Gly Gly Gly Tyr Gly Trp Phe Leu Gly Gln Asn Gln Gly

100 105 110

Gly Ala Asn Gly Ala Tyr Asn Ile Val Lys Ile Pro Tyr Ser Ala Thr

115 120 125

Tyr Pro Thr Ala Phe Arg Pro Lys Val Phe Lys Asn Ala Val Thr Val

130 135 140

Thr Gly Asn Ile Gly Leu Asn Lys Gly Asp Tyr Phe Ile Asp Val Ser

145 150 155 160

Ala Tyr Gln Gln Ala Asp Leu Thr Thr Thr Cys Gln Gln Ala Gly Thr

165 170 175

Thr Lys Thr Ile Ile Lys Val Ser Glu Ser Ile Ala Trp Leu Ser Asp

180 185 190

Arg His Gln Gln Gln Ala Asn Thr Ser Asp Pro Ile Gly Tyr Tyr His

195 200 205

Phe Gly Arg Phe Gly Gly Asp Ser Ala Leu Ala Gln Arg Glu Ala Asp

210 215 220

Leu Phe Leu Ser Asn Leu Pro Ser Lys Lys Val Ser Tyr Leu Val Ile

225 230 235 240

Asp Tyr Glu Asp Ser Ala Ser Ala Asp Lys Gln Ala Asn Thr Asn Ala

245 250 255

Val Ile Ala Phe Met Asp Lys Ile Ala Ser Ala Gly Tyr Lys Pro Ile

260 265 270

Tyr Tyr Ser Tyr Lys Pro Phe Thr Leu Asn Asn Ile Asp Tyr Gln Lys

275 280 285

Ile Ile Ala Lys Tyr Pro Asn Ser Ile Trp Ile Ala Gly Tyr Pro Asp

290 295 300

Tyr Glu Val Arg Thr Glu Pro Leu Trp Glu Phe Phe Pro Ser Met Asp

305 310 315 320

Gly Val Arg Trp Trp Gln Phe Thr Ser Val Gly Val Ala Gly Gly Leu

325 330 335

Asp Lys Asn Ile Val Leu Leu Ala Asp Asp Ser Ser Lys Met Asp Ile

340 345 350

Pro Lys Val Asp Lys Pro Gln Glu Leu Thr Phe Tyr Gln Lys Leu Ala

355 360 365

Thr Asn Thr Lys Leu Asp Asn Ser Asn Val Pro Tyr Tyr Glu Ala Thr

370 375 380

Leu Ser Thr Asp Tyr Tyr Val Glu Ser Lys Pro Asn Ala Ser Ser Ala

385 390 395 400

Asp Lys Glu Phe Ile Lys Ala Gly Thr Arg Val Arg Val Tyr Glu Lys

405 410 415

Val Asn Gly Trp Ser Arg Ile Asn His Pro Glu Ser Ala Gln Trp Val

420 425 430

Glu Asp Ser Tyr Leu Val Asn Ala Thr Asp Met

435 440

<210> 68

<211> 334

<212> PROTEIN

<213> (Phi)FWLLm3

<400> 68

Met Val Lys Tyr Thr Val Glu Asn Lys Ile Ile Ala Gly Leu Pro Lys

1 5 10 15

Gly Lys Leu Lys Gly Ala Asn Phe Val Ile Ala His Glu Thr Ala Asn

20 25 30

Ser Lys Ser Thr Ile Asp Asn Glu Val Ser Tyr Met Thr Arg Asn Trp

35 40 45

Gln Asn Ala Phe Val Thr His Phe Val Gly Gly Gly Gly Arg Val Val

50 55 60

Gln Val Ala Asn Val Asn Tyr Val Ser Trp Gly Ala Gly Gln Tyr Ala

65 70 75 80

Asn Ser Tyr Ser Tyr Ala Gln Val Glu Leu Cys Arg Thr Ser Asn Ala

85 90 95

Thr Thr Phe Lys Lys Asp Tyr Glu Val Tyr Cys Gln Leu Leu Val Asp

100 105 110

Leu Ala Lys Lys Ala Gly Ile Pro Ile Thr Leu Asp Ser Gly Ser Lys

115 120 125

Thr Ser Asp Lys Gly Ile Lys Ser His Lys Trp Val Ala Asp Lys Leu

130 135 140

Gly Gly Thr Thr His Gln Asp Pro Tyr Ala Tyr Leu Ser Ser Trp Gly

145 150 155 160

Ile Ser Lys Ala Gln Phe Ala Ser Asp Leu Ala Lys Val Ser Gly Gly

165 170 175

Gly Asn Thr Gly Thr Ala Pro Ala Lys Pro Ser Thr Pro Ser Thr Asn

180 185 190

Leu Asp Lys Leu Gly Leu Val Asp Tyr Met Asn Ala Lys Lys Met Asp

195 200 205

Ser Ser Tyr Ser Asn Arg Ala Lys Leu Ala Lys Gln Tyr Gly Ile Ala

210 215 220

Asn Tyr Ser Gly Thr Ala Ser Gln Asn Thr Thr Leu Leu Ser Lys Ile

225 230 235 240

Lys Gly Gly Ala Pro Lys Pro Ser Thr Pro Ala Pro Lys Pro Ser Thr

245 250 255

Ser Thr Ala Lys Lys Ile Tyr Phe Pro Pro Asn Lys Gly Asn Trp Ser

260 265 270

Val Tyr Pro Thr Asn Lys Ala Pro Val Lys Ala Asn Ala Ile Gly Ala

275 280 285

Ile Asn Pro Thr Lys Phe Gly Gly Leu Thr Tyr Thr Ile Gln Lys Asp

290 295 300

Arg Gly Asn Gly Val Tyr Glu Ile Gln Thr Asp Gln Phe Gly Arg Val

305 310 315 320

Gln Val Tyr Gly Ala Pro Ser Thr Gly Ala Val Ile Lys Lys

325 330

<210> 69

<211> 1278

<212> PROTEIN

<213> (Phi)BPS13

<400> 69

Met Ala Lys Arg Glu Lys Tyr Ile Phe Asp Val Glu Ala Glu Val Gly

1 5 10 15

Lys Ala Ala Lys Ser Ile Lys Ser Leu Glu Ala Glu Leu Ser Lys Leu

20 25 30

Gln Lys Leu Asn Lys Glu Ile Asp Ala Thr Gly Gly Asp Arg Thr Glu

35 40 45

Lys Glu Met Leu Ala Thr Leu Lys Ala Ala Lys Glu Val Asn Ala Glu

50 55 60

Tyr Gln Lys Met Gln Arg Ile Leu Lys Asp Leu Ser Lys Tyr Ser Gly

65 70 75 80

Lys Val Ser Arg Lys Glu Phe Asn Asp Ser Lys Val Ile Asn Asn Ala

85 90 95

Lys Thr Ser Val Gln Gly Gly Lys Val Thr Asp Ser Phe Gly Gln Met

100 105 110

Leu Lys Asn Met Glu Arg Gln Ile Asn Ser Val Asn Lys Gln Phe Asp

115 120 125

Asn His Arg Lys Ala Met Val Asp Arg Gly Gln Gln Tyr Thr Pro His

130 135 140

Leu Lys Thr Asn Arg Lys Asp Ser Gln Gly Asn Ser Asn Pro Ser Met

145 150 155 160

Met Gly Arg Asn Lys Ser Thr Thr Gln Asp Met Glu Lys Ala Val Asp

165 170 175

Lys Phe Leu Asn Gly Gln Asn Glu Ala Thr Thr Gly Leu Asn Gln Ala

180 185 190

Leu Tyr Gln Leu Lys Glu Ile Ser Lys Leu Asn Arg Arg Ser Glu Ser

195 200 205

Leu Ser Arg Arg Ala Ser Ala Ser Gly Tyr Met Ser Phe Gln Gln Tyr

210 215 220

Ser Asn Phe Thr Gly Asp Arg Arg Thr Val Gln Gln Thr Tyr Gly Gly

225 230 235 240

Leu Lys Thr Ala Asn Arg Glu Arg Val Leu Glu Leu Ser Gly Gln Ala

245 250 255

Thr Gly Ile Ser Lys Glu Leu Asp Arg Leu Asn Ser Lys Lys Gly Leu

260 265 270

Thr Ala Arg Glu Gly Glu Glu Arg Lys Lys Leu Met Arg Gln Leu Glu

275 280 285

Gly Ile Asp Ala Glu Leu Thr Ala Arg Lys Lys Leu Asn Ser Ser Leu

290 295 300

Asp Glu Thr Thr Ser Asn Met Glu Lys Phe Asn Gln Ser Leu Val Asp

305 310 315 320

Ala Gln Val Ser Val Lys Pro Glu Arg Gly Thr Met Arg Gly Met Met

325 330 335

Tyr Glu Arg Ala Pro Ala Ile Ala Leu Ala Ile Gly Gly Ala Ile Thr

340 345 350

Ala Thr Ile Gly Lys Leu Tyr Ser Glu Gly Gly Asn His Ser Lys Ala

355 360 365

Met Arg Pro Asp Glu Met Tyr Val Gly Gln Gln Thr Gly Ala Val Gly

370 375 380

Ala Asn Trp Arg Pro Asn Arg Thr Ala Thr Met Arg Ser Gly Leu Gly

385 390 395 400

Asn His Leu Gly Phe Thr Gly Gln Glu Met Met Glu Phe Gln Ser Asn

405 410 415

Tyr Leu Ser Ala Asn Gly Tyr His Gly Ala Glu Asp Met Lys Ala Ala

420 425 430

Thr Thr Gly Gln Ala Thr Phe Ala Arg Ala Thr Gly Leu Gly Ser Asp

435 440 445

Glu Val Lys Asp Phe Phe Asn Thr Ala Tyr Arg Ser Gly Gly Ile Asp

450 455 460

Gly Asn Gln Thr Lys Gln Phe Gln Asn Ala Phe Leu Gly Ala Met Lys

465 470 475 480

Gln Ser Gly Ala Val Gly Arg Glu Lys Asp Gln Leu Lys Ala Leu Asn

485 490 495

Gly Ile Leu Ser Ser Met Ser Gln Asn Arg Thr Val Ser Asn Gln Asp

500 505 510

Met Met Arg Thr Val Gly Leu Gln Ser Ala Ile Ser Ser Ser Gly Val

515 520 525

Ala Ser Leu Gln Gly Thr Lys Gly Gly Ala Leu Met Glu Gln Leu Asp

530 535 540

Asn Gly Ile Arg Glu Gly Phe Asn Asp Pro Gln Met Arg Val Leu Phe

545 550 555 560

Gly Gln Gly Thr Lys Tyr Gln Gly Met Gly Gly Arg Ala Ala Leu Arg

565 570 575

Lys Gln Met Glu Lys Gly Ile Ser Asp Pro Asp Asn Leu Asn Thr Leu

580 585 590

Ile Asp Ala Ser Lys Ala Ser Ala Gly Gln Asp Pro Ala Glu Gln Ala

595 600 605

Glu Val Leu Ala Thr Leu Ala Ser Lys Met Gly Val Asn Met Ser Ser

610 615 620

Asp Gln Ala Arg Gly Leu Ile Asp Leu Gln Pro Ser Gly Lys Leu Thr

625 630 635 640

Lys Glu Asn Ile Asp Lys Val Met Lys Glu Gly Leu Lys Glu Gly Ser

645 650 655

Ile Glu Ser Ala Lys Arg Asp Lys Ala Tyr Ser Glu Ser Lys Ala Ser

660 665 670

Ile Asp Asn Ser Ser Glu Ala Ala Thr Ala Lys Gln Ala Thr Glu Leu

675 680 685

Asn Asp Met Gly Ser Lys Leu Arg Gln Ala Asn Ala Ala Leu Gly Gly

690 695 700

Leu Pro Ala Pro Leu Tyr Thr Ala Ile Ala Ala Val Val Ala Phe Thr

705 710 715 720

Ala Ala Val Ala Gly Ser Ala Leu Met Phe Lys Gly Ala Ser Trp Leu

725 730 735

Lys Gly Gly Met Ala Ser Lys Tyr Gly Gly Lys Gly Gly Lys Gly Gly

740 745 750

Lys Gly Gly Gly Thr Gly Gly Gly Gly Gly Ala Gly Gly Ala Ala Ala

755 760 765

Thr Gly Ala Gly Ala Ala Ala Gly Ala Gly Gly Val Gly Ala Ala Ala

770 775 780

Ala Gly Glu Val Gly Ala Gly Val Ala Ala Gly Gly Ala Ala Ala Gly

785 790 795 800

Ala Ala Ala Gly Gly Ser Lys Leu Ala Gly Val Gly Lys Gly Phe Met

805 810 815

Lys Gly Ala Gly Lys Leu Met Leu Pro Leu Gly Ile Leu Met Gly Ala

820 825 830

Ser Glu Ile Met Gln Ala Pro Glu Glu Ala Lys Gly Ser Ala Ile Gly

835 840 845

Ser Ala Val Gly Gly Ile Gly Gly Gly Ile Ala Gly Gly Ala Ala Thr

850 855 860

Gly Ala Ile Ala Gly Ser Phe Leu Gly Pro Ile Gly Thr Ala Val Gly

865 870 875 880

Gly Ile Ala Gly Gly Ile Ala Gly Gly Phe Ala Gly Ser Ser Leu Gly

885 890 895

Glu Thr Ile Gly Gly Trp Phe Asp Ser Gly Pro Lys Glu Asp Ala Ser

900 905 910

Ala Ala Asp Lys Ala Lys Ala Asp Ala Ser Ala Ala Ala Leu Ala Ala

915 920 925

Ala Ala Gly Thr Ser Gly Ala Val Gly Ser Ser Ala Leu Gln Ser Gln

930 935 940

Met Ala Gln Gly Ile Thr Gly Ala Pro Asn Met Ser Gln Val Gly Ser

945 950 955 960

Met Ala Ser Ala Leu Gly Ile Ser Ser Gly Ala Met Ala Ser Ala Leu

965 970 975

Gly Ile Ser Ser Gly Gln Glu Asn Gln Ile Gln Thr Met Thr Asp Lys

980 985 990

Glu Asn Thr Asn Thr Lys Lys Ala Asn Glu Ala Lys Lys Gly Asp Asn

995 1000 1005

Leu Ser Tyr Glu Arg Glu Asn Ile Ser Met Tyr Glu Arg Val Leu

1010 1015 1020

Thr Arg Ala Glu Gln Ile Leu Ala Gln Ala Arg Ala Gln Asn Gly

1025 1030 1035

Ile Met Gly Val Gly Gly Gly Gly Thr Ala Gly Ala Gly Gly Gly

1040 1045 1050

Ile Asn Gly Phe Thr Gly Gly Gly Lys Leu Gln Phe Leu Ala Ala

1055 1060 1065

Gly Gln Lys Trp Ser Ser Ser Asn Leu Gln Gln His Asp Leu Gly

1070 1075 1080

Phe Thr Asp Gln Asn Leu Thr Ala Glu Asp Leu Asp Lys Trp Ile

1085 1090 1095

Asp Ser Lys Ala Pro Gly Ser Met Met Arg Gly Met Gly Ala

1100 1105 1110

Thr Phe Leu Lys Ala Gly Gln Glu Tyr Gly Leu Asp Pro Arg Tyr

1115 1120 1125

Leu Ile Ala His Ala Ala Glu Glu Ser Gly Trp Gly Thr Ser Lys

1130 1135 1140

Ile Ala Arg Asp Lys Gly Asn Phe Phe Gly Ile Gly Ala Phe Asp

1145 1150 1155

Asp Ser Pro Tyr Ser Ser Ala Tyr Glu Phe Lys Asp Gly Thr Gly

1160 1165 1170

Ser Ala Ala Glu Arg Gly Ile Met Gly Gly Ala Lys Trp Ile Ser

1175 1180 1185

Glu Lys Tyr Tyr Gly Lys Gly Asn Thr Thr Leu Asp Lys Met Lys

1190 1195 1200

Ala Ala Gly Tyr Ala Thr Asn Ala Ser Trp Ala Pro Asn Ile Ala

1205 1210 1215

Ser Ile Met Ala Gly Ala Pro Thr Gly Ser Gly Ser Gly Asn Val

1220 1225 1230

Thr Ala Thr Ile Asn Val Asn Val Lys Gly Asp Glu Lys Val Ser

1235 1240 1245

Asp Lys Leu Lys Asn Ser Ser Asp Met Lys Lys Ala Gly Lys Asp

1250 1255 1260

Ile Gly Ser Leu Leu Gly Phe Tyr Ser Arg Glu Met Thr Ile Ala

1265 1270 1275

<210> 70

<211> 495

<212> PROTEIN

<213> (Phi)GH15

<400> 70

Met Ala Lys Thr Gln Ala Glu Ile Asn Lys Arg Leu Asp Ala Tyr Ala

1 5 10 15

Lys Gly Thr Val Asp Ser Pro Tyr Arg Ile Lys Lys Ala Thr Ser Tyr

20 25 30

Asp Pro Ser Phe Gly Val Met Glu Ala Gly Ala Ile Asp Ala Asp Gly

35 40 45

Tyr Tyr His Ala Gln Cys Gln Asp Leu Ile Thr Asp Tyr Val Leu Trp

50 55 60

Leu Thr Asp Asn Lys Val Arg Thr Trp Gly Asn Ala Lys Asp Gln Ile

65 70 75 80

Lys Gln Ser Tyr Gly Thr Gly Phe Lys Ile His Glu Asn Lys Pro Ser

85 90 95

Thr Val Pro Lys Lys Gly Trp Ile Ala Val Phe Thr Ser Gly Ser Tyr

100 105 110

Gln Gln Trp Gly His Ile Gly Ile Val Tyr Asp Gly Gly Asn Thr Ser

115 120 125

Thr Phe Thr Ile Leu Glu Gln Asn Trp Asn Gly Tyr Ala Asn Lys Lys

130 135 140

Pro Thr Lys Arg Val Asp Asn Tyr Tyr Gly Leu Thr His Phe Ile Glu

145 150 155 160

Ile Pro Val Lys Ala Gly Thr Thr Val Lys Lys Glu Thr Ala Lys Lys

165 170 175

Ser Ala Ser Lys Thr Pro Ala Pro Lys Lys Lys Ala Thr Leu Lys Val

180 185 190

Ser Lys Asn His Ile Asn Tyr Thr Met Asp Lys Arg Gly Lys Lys Pro

195 200 205

Glu Gly Met Val Ile His Asn Asp Ala Gly Arg Ser Ser Gly Gln Gln

210 215 220

Tyr Glu Asn Ser Leu Ala Asn Ala Gly Tyr Ala Arg Tyr Ala Asn Gly

225 230 235 240

Ile Ala His Tyr Tyr Gly Ser Glu Gly Tyr Val Trp Glu Ala Ile Asp

245 250 255

Ala Lys Asn Gln Ile Ala Trp His Thr Gly Asp Gly Thr Gly Ala Asn

260 265 270

Ser Gly Asn Phe Arg Phe Ala Gly Ile Glu Val Cys Gln Ser Met Ser

275 280 285

Ala Ser Asp Ala Gln Phe Leu Lys Asn Glu Gln Ala Val Phe Gln Phe

290 295 300

Thr Ala Glu Lys Phe Lys Glu Trp Gly Leu Thr Pro Asn Arg Lys Thr

305 310 315 320

Val Arg Leu His Met Glu Phe Val Pro Thr Ala Cys Pro His Arg Ser

325 330 335

Met Val Leu His Thr Gly Phe Asn Pro Val Thr Gln Gly Arg Pro Ser

340 345 350

Gln Ala Ile Met Asn Lys Leu Lys Asp Tyr Phe Ile Lys Gln Ile Lys

355 360 365

Asn Tyr Met Asp Lys Gly Thr Ser Ser Ser Thr Val Val Lys Asp Gly

370 375 380

Lys Thr Ser Ser Ala Ser Thr Pro Ala Thr Arg Pro Val Thr Gly Ser

385 390 395 400

Trp Lys Lys Asn Gln Tyr Gly Thr Trp Tyr Lys Pro Glu Asn Ala Thr

405 410 415

Phe Val Asn Gly Asn Gln Pro Ile Val Thr Arg Ile Gly Ser Pro Phe

420 425 430

Leu Asn Ala Pro Val Gly Gly Asn Leu Pro Ala Gly Ala Thr Ile Val

435 440 445

Tyr Asp Glu Val Cys Ile Gln Ala Gly His Ile Trp Ile Gly Tyr Asn

450 455 460

Ala Tyr Asn Gly Asp Arg Val Tyr Cys Pro Val Arg Thr Cys Gln Gly

465 470 475 480

Val Pro Pro Asn His Ile Pro Gly Val Ala Trp Gly Val Phe Lys

485 490 495

<210> 71

<211> 264

<212> PROTEIN

<213> (Phi)8074-B1

<400> 71

Met Lys Ile Gly Ile Asp Met Gly His Thr Leu Ser Gly Ala Asp Tyr

1 5 10 15

Gly Val Val Gly Leu Arg Pro Glu Ser Val Leu Thr Arg Glu Val Gly

20 25 30

Thr Lys Val Ile Tyr Lys Leu Gln Lys Leu Gly His Val Val Val Asn

35 40 45

Cys Thr Val Asp Lys Ala Ser Ser Val Ser Glu Ser Leu Tyr Thr Arg

50 55 60

Tyr Tyr Arg Ala Asn Gln Ala Asn Val Asp Leu Phe Ile Ser Ile His

65 70 75 80

Phe Asn Ala Thr Pro Gly Gly Thr Gly Thr Glu Val Tyr Thr Tyr Ala

85 90 95

Gly Arg Gln Leu Gly Glu Ala Thr Arg Ile Arg Gln Glu Phe Lys Ser

100 105 110

Leu Gly Leu Arg Asp Arg Gly Thr Lys Asp Gly Ser Gly Leu Ala Val

115 120 125

Ile Arg Asn Thr Lys Ala Lys Ala Met Leu Val Glu Cys Cys Phe Cys

130 135 140

Asp Asn Pro Asn Asp Met Lys Leu Tyr Asn Ser Glu Ser Phe Ser Asn

145 150 155 160

Ala Ile Val Lys Gly Ile Thr Gly Lys Leu Pro Asn Gly Glu Ser Gly

165 170 175

Asn Asn Asn Gln Gly Gly Asn Lys Val Lys Ala Val Val Ile Tyr Asn

180 185 190

Glu Gly Ala Asp Arg Arg Gly Ala Glu Tyr Leu Ala Asp Tyr Leu Asn

195 200 205

Cys Pro Thr Ile Ser Asn Ser Arg Thr Phe Asp Tyr Ser Cys Val Glu

210 215 220

His Val Tyr Ala Val Gly Gly Lys Lys Glu Gln Tyr Thr Lys Tyr Leu

225 230 235 240

Lys Thr Leu Leu Ser Gly Ala Asn Arg Tyr Asp Thr Met Gln Gln Ile

245 250 255

Leu Asn Phe Ile Asn Gly Gly Lys

260

<210> 72

<211> 209

<212> PROTEIN

<213> (Phi)SPN1S

<400> 72

Met Asp Ile Asn Gln Phe Arg Arg Ala Ser Gly Ile Asn Glu Gln Leu

1 5 10 15

Ala Ala Arg Trp Phe Pro His Ile Thr Thr Ala Met Asn Glu Phe Gly

20 25 30

Ile Thr Lys Pro Asp Asp Gln Ala Met Phe Ile Ala Gln Val Gly His

35 40 45

Glu Ser Gly Gly Phe Thr Arg Leu Gln Glu Asn Phe Asn Tyr Ser Val

50 55 60

Asn Gly Leu Ser Gly Phe Ile Arg Ala Gly Arg Ile Thr Pro Asp Gln

65 70 75 80

Ala Asn Ala Leu Gly Arg Lys Thr Tyr Glu Lys Ser Leu Pro Leu Glu

85 90 95

Arg Gln Arg Ala Ile Ala Asn Leu Val Tyr Ser Lys Arg Met Gly Asn

100 105 110

Asn Gly Pro Gly Asp Gly Trp Asn Tyr Arg Gly Arg Gly Leu Ile Gln

115 120 125

Ile Thr Gly Leu Asn Asn Tyr Arg Asp Cys Gly Asn Gly Leu Lys Val

130 135 140

Asp Leu Val Ala Gln Pro Glu Leu Leu Ala Gln Asp Glu Tyr Ala Ala

145 150 155 160

Arg Ser Ala Ala Trp Phe Phe Ser Ser Lys Gly Cys Met Lys Tyr Thr

165 170 175

Gly Asp Leu Val Arg Val Thr Gln Ile Ile Asn Gly Gly Gln Asn Gly

180 185 190

Ile Asp Asp Arg Arg Thr Arg Tyr Ala Ala Ala Arg Lys Val Leu Ala

195 200 205

Leu

<210> 73

<211> 290

<212> PROTEIN

<213> (Phi)CN77

<400> 73

Met Gly Tyr Trp Gly Tyr Pro Asn Gly Gln Ile Pro Asn Asp Lys Met

1 5 10 15

Ala Leu Tyr Arg Gly Cys Leu Leu Arg Ala Asp Ala Ala Ala Gln Ala

20 25 30

Tyr Ala Leu Gln Asp Ala Tyr Thr Arg Ala Thr Gly Lys Pro Leu Val

35 40 45

Ile Leu Glu Gly Tyr Arg Asp Leu Thr Arg Gln Lys Tyr Leu Arg Asn

50 55 60

Leu Tyr Leu Ser Gly Arg Gly Asn Ile Ala Ala Val Pro Gly Leu Ser

65 70 75 80

Asn His Gly Trp Gly Leu Ala Cys Asp Phe Ala Ala Pro Leu Asn Ser

85 90 95

Ser Gly Ser Glu Glu His Arg Trp Met Arg Gln Asn Ala Pro Leu Phe

100 105 110

Gly Phe Asp Trp Ala Arg Gly Lys Ala Asp Asn Glu Pro Trp His Trp

115 120 125

Glu Tyr Gly Asn Val Pro Val Ser Arg Trp Ala Ser Leu Asp Val Thr

130 135 140

Pro Ile Asp Arg Asn Asp Met Ala Asp Ile Thr Glu Gly Gln Met Gln

145 150 155 160

Arg Ile Ala Val Ile Leu Leu Asp Thr Glu Ile Gln Thr Pro Leu Gly

165 170 175

Pro Arg Leu Val Lys His Ala Leu Gly Asp Ala Leu Leu Leu Gly Gln

180 185 190

Ala Asn Ala Asn Ser Ile Ala Glu Val Pro Asp Lys Thr Trp Asp Val

195 200 205

Leu Val Asp His Pro Leu Ala Lys Asn Glu Asp Gly Thr Pro Leu Lys

210 215 220

Val Arg Leu Gly Asp Val Ala Lys Tyr Glu Pro Leu Glu His Gln Asn

225 230 235 240

Thr Arg Asp Ala Ile Ala Lys Leu Gly Thr Leu Gln Phe Thr Asp Lys

245 250 255

Gln Leu Ala Thr Ile Gly Ala Gly Val Lys Pro Ile Asp Glu Ala Ser

260 265 270

Leu Val Lys Lys Ile Val Asp Gly Val Arg Ala Leu Phe Gly Arg Ala

275 280 285

Ala Ala

290

<210> 74

<211> 185

<212> PROTEIN

<213> (Phi)AB2

<400> 74

Met Ile Leu Thr Lys Asp Gly Phe Ser Ile Ile Arg Asn Glu Leu Phe

1 5 10 15

Gly Gly Lys Leu Asp Gln Thr Gln Val Asp Ala Ile Asn Phe Ile Val

20 25 30

Ala Lys Ala Thr Glu Ser Gly Leu Thr Tyr Pro Glu Ala Ala Tyr Leu

35 40 45

Leu Ala Thr Ile Tyr His Glu Thr Gly Leu Pro Ser Gly Tyr Arg Thr

50 55 60

Met Gln Pro Ile Lys Glu Ala Gly Ser Asp Ser Tyr Leu Arg Ser Lys

65 70 75 80

Lys Tyr Tyr Pro Tyr Ile Gly Tyr Gly Tyr Val Gln Leu Thr Trp Lys

85 90 95

Glu Asn Tyr Glu Arg Ile Gly Lys Leu Ile Gly Val Asp Leu Ile Lys

100 105 110

Asn Pro Glu Lys Ala Leu Glu Pro Leu Ile Ala Ile Gln Ile Ala Ile

115 120 125

Lys Gly Met Leu Asn Gly Trp Phe Thr Gly Val Gly Phe Arg Arg Lys

130 135 140

Arg Pro Val Ser Lys Tyr Asn Lys Gln Gln Tyr Val Ala Ala Arg Asn

145 150 155 160

Ile Ile Asn Gly Lys Asp Lys Ala Glu Leu Ile Ala Lys Tyr Ala Ile

165 170 175

Ile Phe Glu Arg Ala Leu Arg Ser Leu

180 185

<210> 75

<211> 262

<212> PROTEIN

<213> (Phi)B4

<400> 75

Met Ala Met Ala Leu Gln Thr Leu Ile Asp Lys Ala Asn Arg Lys Leu

1 5 10 15

Asn Val Ser Gly Met Arg Lys Asp Val Ala Asp Arg Thr Arg Ala Val

20 25 30

Ile Thr Gln Met His Ala Gln Gly Ile Tyr Ile Cys Val Ala Gln Gly

35 40 45

Phe Arg Ser Phe Ala Glu Gln Asn Ala Leu Tyr Ala Gln Gly Arg Thr

50 55 60

Lys Pro Gly Ser Ile Val Thr Asn Ala Arg Gly Gly Gln Ser Asn His

65 70 75 80

Asn Tyr Gly Val Ala Val Asp Leu Cys Leu Tyr Thr Gln Asp Gly Ser

85 90 95

Asp Val Ile Trp Thr Val Glu Gly Asn Phe Arg Lys Val Ile Ala Ala

100 105 110

Met Lys Ala Gln Gly Phe Lys Trp Gly Gly Asp Trp Val Ser Phe Lys

115 120 125

Asp Tyr Pro His Phe Glu Leu Tyr Asp Val Val Gly Gly Gln Lys Pro

130 135 140

Pro Ala Asp Asn Gly Gly Ala Val Asp Asn Gly Gly Gly Ser Gly Ser

145 150 155 160

Thr Gly Gly Ser Gly Gly Gly Ser Thr Gly Gly Gly Ser Thr Gly Gly

165 170 175

Gly Tyr Asp Ser Ser Trp Phe Thr Lys Glu Thr Gly Thr Phe Val Thr

180 185 190

Asn Thr Ser Ile Lys Leu Arg Thr Ala Pro Phe Thr Ser Ala Asp Val

195 200 205

Ile Ala Thr Leu Pro Ala Gly Ser Pro Val Asn Tyr Asn Gly Phe Gly

210 215 220

Ile Glu Tyr Asp Gly Tyr Val Trp Ile Arg Gln Pro Arg Ser Asn Gly

225 230 235 240

Tyr Gly Tyr Leu Ala Thr Gly Glu Ser Lys Gly Gly Lys Arg Gln Asn

245 250 255

Tyr Trp Gly Thr Phe Lys

260

<210> 76

<211> 274

<212> PROTEIN

<213> (Phi)CTP1

<400> 76

Met Lys Lys Ile Ala Asp Ile Ser Asn Leu Asn Gly Asn Val Asp Val

1 5 10 15

Lys Leu Leu Phe Asn Leu Gly Tyr Ile Gly Ile Ile Ala Lys Ala Ser

20 25 30

Glu Gly Gly Thr Phe Val Asp Lys Tyr Tyr Lys Gln Asn Tyr Thr Asn

35 40 45

Thr Lys Ala Gln Gly Lys Ile Thr Gly Ala Tyr His Phe Ala Asn Phe

50 55 60

Ser Thr Ile Ala Lys Ala Gln Gln Glu Ala Asn Phe Phe Leu Asn Cys

65 70 75 80

Ile Ala Gly Thr Thr Pro Asp Phe Val Val Leu Asp Leu Glu Gln Gln

85 90 95

Cys Thr Gly Asp Ile Thr Asp Ala Cys Leu Ala Phe Leu Asn Ile Val

100 105 110

Ala Lys Lys Phe Lys Cys Val Val Tyr Cys Asn Ser Ser Phe Ile Lys

115 120 125

Glu His Leu Asn Ser Lys Ile Cys Ala Tyr Pro Leu Trp Ile Ala Asn

130 135 140

Tyr Gly Val Ala Thr Pro Ala Phe Thr Leu Trp Thr Lys Tyr Ala Met

145 150 155 160

Trp Gln Phe Thr Glu Lys Gly Gln Val Ser Gly Ile Ser Gly Tyr Ile

165 170 175

Asp Phe Ser Tyr Ile Thr Asp Glu Phe Ile Lys Tyr Ile Lys Gly Glu

180 185 190

Asp Glu Val Glu Asn Leu Val Val Tyr Asn Asp Gly Ala Asp Gln Arg

195 200 205

Ala Ala Glu Tyr Leu Ala Asp Arg Leu Ala Cys Pro Thr Ile Asn Asn

210 215 220

Ala Arg Lys Phe Asp Tyr Ser Asn Val Lys Asn Val Tyr Ala Val Gly

225 230 235 240

Gly Asn Lys Glu Gln Tyr Thr Ser Tyr Leu Thr Thr Leu Ile Ala Gly

245 250 255

Ser Thr Arg Tyr Thr Thr Met Gln Ala Val Leu Asp Tyr Ile Lys Asn

260 265 270

Leu Lys

<210> 77

<211> 628

<212> PROTEIN

<213> Staphylococcus virus 187

<400> 77

Met Ala Leu Pro Lys Thr Gly Lys Pro Thr Ala Lys Gln Val Val Asp

1 5 10 15

Trp Ala Ile Asn Leu Ile Gly Ser Gly Val Asp Val Asp Gly Tyr Tyr

20 25 30

Gly Arg Gln Cys Trp Asp Leu Pro Asn Tyr Ile Phe Asn Arg Tyr Trp

35 40 45

Asn Phe Lys Thr Pro Gly Asn Ala Arg Asp Met Ala Trp Tyr Arg Tyr

50 55 60

Pro Glu Gly Phe Lys Val Phe Arg Asn Thr Ser Asp Phe Val Pro Lys

65 70 75 80

Pro Gly Asp Ile Ala Val Trp Thr Gly Gly Asn Tyr Asn Trp Asn Thr

85 90 95

Trp Gly His Thr Gly Ile Val Val Gly Pro Ser Thr Lys Ser Tyr Phe

100 105 110

Tyr Ser Val Asp Gln Asn Trp Asn Asn Ser Asn Ser Tyr Val Gly Ser

115 120 125

Pro Ala Ala Lys Ile Lys His Ser Tyr Phe Gly Val Thr His Phe Val

130 135 140

Arg Pro Ala Tyr Lys Ala Glu Pro Lys Pro Thr Pro Pro Ala Gln Asn

145 150 155 160

Asn Pro Ala Pro Lys Asp Pro Glu Pro Ser Lys Lys Pro Glu Ser Asn

165 170 175

Lys Pro Ile Tyr Lys Val Val Thr Lys Ile Leu Phe Thr Thr Ala His

180 185 190

Ile Glu His Val Lys Ala Asn Arg Phe Val His Tyr Ile Thr Lys Ser

195 200 205

Asp Asn His Asn Asn Lys Pro Asn Lys Ile Val Ile Lys Asn Thr Asn

210 215 220

Thr Ala Leu Ser Thr Ile Asp Val Tyr Arg Tyr Arg Asp Glu Leu Asp

225 230 235 240

Lys Asp Glu Ile Pro His Phe Phe Val Asp Arg Leu Asn Val Trp Ala

245 250 255

Cys Arg Pro Ile Glu Asp Ser Ile Asn Gly Tyr His Asp Ser Val Val

260 265 270

Leu Ser Ile Thr Glu Thr Arg Thr Ala Leu Ser Asp Asn Phe Lys Met

275 280 285

Asn Glu Ile Glu Cys Leu Ser Leu Ala Glu Ser Ile Leu Lys Ala Asn

290 295 300

Asn Lys Lys Met Ser Ala Ser Asn Ile Ile Val Asp Asn Lys Ala Trp

305 310 315 320

Arg Thr Phe Lys Leu His Thr Gly Lys Asp Ser Leu Lys Ser Ser Ser

325 330 335

Phe Thr Ser Lys Asp Tyr Gln Lys Ala Val Asn Glu Leu Ile Lys Leu

340 345 350

Phe Asn Asp Lys Asp Lys Leu Leu Asn Asn Lys Pro Lys Asp Val Val

355 360 365

Glu Arg Ile Arg Ile Arg Thr Ile Val Lys Glu Asn Thr Lys Phe Val

370 375 380

Pro Ser Glu Leu Lys Pro Arg Asn Asn Ile Arg Asp Lys Gln Asp Ser

385 390 395 400

Lys Ile Asp Arg Val Ile Asn Asn Tyr Thr Leu Lys Gln Ala Leu Asn

405 410 415

Ile Gln Tyr Lys Leu Asn Pro Lys Pro Gln Thr Ser Asn Gly Val Ser

420 425 430

Trp Tyr Asn Ala Ser Val Asn Gln Ile Lys Ser Ala Met Asp Thr Thr

435 440 445

Lys Ile Phe Asn Asn Asn Val Gln Val Tyr Gln Phe Leu Lys Leu Asn

450 455 460

Gln Tyr Gln Gly Ile Pro Val Asp Lys Leu Asn Lys Leu Leu Val Gly

465 470 475 480

Lys Gly Thr Leu Ala Asn Gln Gly His Ala Phe Ala Asp Gly Cys Lys

485 490 495

Lys Tyr Asn Ile Asn Glu Ile Tyr Leu Ile Ala His Arg Phe Leu Glu

500 505 510

Ser Ala Asn Gly Thr Ser Phe Phe Ala Ser Gly Lys Thr Gly Val Tyr

515 520 525

Asn Tyr Phe Gly Ile Gly Ala Phe Asp Asn Asn Pro Asn Asn Ala Met

530 535 540

Ala Phe Ala Arg Ser His Gly Trp Thr Ser Pro Thr Lys Ala Ile Ile

545 550 555 560

Gly Gly Ala Glu Phe Val Gly Lys Gly Tyr Phe Asn Val Gly Gln Asn

565 570 575

Thr Leu Tyr Arg Met Arg Trp Asn Pro Gln Lys Pro Gly Thr His Gln

580 585 590

Tyr Ala Thr Asp Ile Ser Trp Ala Lys Val Gln Ala Gln Met Ile Ser

595 600 605

Ala Met Tyr Lys Glu Ile Gly Leu Thr Gly Asp Tyr Phe Ile Tyr Asp

610 615 620

Gln Tyr Lys Lys

625

<210> 78

<211> 291

<212> PROTEIN

<213> (Phi)P35

<400> 78

Met Ala Arg Lys Phe Thr Lys Ala Glu Leu Val Ala Lys Ala Glu Lys

1 5 10 15

Lys Val Gly Gly Leu Lys Pro Asp Val Lys Lys Ala Val Leu Ser Ala

20 25 30

Val Lys Glu Ala Tyr Asp Arg Tyr Gly Ile Gly Ile Ile Val Ser Gln

35 40 45

Gly Tyr Arg Ser Ile Ala Glu Gln Asn Gly Leu Tyr Ala Gln Gly Arg

50 55 60

Thr Lys Pro Gly Asn Ile Val Thr Asn Ala Lys Gly Gly Gln Ser Asn

65 70 75 80

His Asn Phe Gly Val Ala Val Asp Phe Ala Ile Asp Leu Ile Asp Asp

85 90 95

Gly Lys Ile Asp Ser Trp Gln Pro Ser Ala Thr Ile Val Asn Met Met

100 105 110

Lys Arg Arg Gly Phe Lys Trp Gly Gly Asp Trp Lys Ser Phe Thr Asp

115 120 125

Leu Pro His Phe Glu Ala Cys Asp Trp Tyr Arg Gly Glu Arg Lys Tyr

130 135 140

Lys Val Asp Thr Ser Glu Trp Lys Lys Lys Glu Asn Ile Asn Ile Val

145 150 155 160

Ile Lys Asp Val Gly Tyr Phe Gln Asp Lys Pro Gln Phe Leu Asn Ser

165 170 175

Lys Ser Val Arg Gln Trp Lys His Gly Thr Lys Val Lys Leu Thr Lys

180 185 190

His Asn Ser His Trp Tyr Thr Gly Val Val Lys Asp Gly Asn Lys Ser

195 200 205

Val Arg Gly Tyr Ile Tyr His Ser Met Ala Lys Val Thr Ser Lys Asn

210 215 220

Ser Asp Gly Ser Val Asn Ala Thr Ile Asn Ala His Ala Phe Cys Trp

225 230 235 240

Asp Asn Lys Lys Leu Asn Gly Gly Asp Phe Ile Asn Leu Lys Arg Gly

245 250 255

Phe Lys Gly Ile Thr His Pro Ala Ser Asp Gly Phe Tyr Pro Leu Tyr

260 265 270

Phe Ala Ser Arg Lys Lys Thr Phe Tyr Ile Pro Arg Tyr Met Phe Asp

275 280 285

Ile Lys Lys

290

<210> 79

<211> 342

<212> PROTEIN

<213> (Phi)CP-7

<400> 79

Met Val Lys Lys Asn Asp Leu Phe Val Asp Val Ala Ser His Gln Gly

1 5 10 15

Tyr Asp Ile Ser Gly Ile Leu Glu Glu Ala Gly Thr Thr Asn Thr Ile

20 25 30

Ile Lys Val Ser Glu Ser Thr Ser Tyr Leu Asn Pro Cys Leu Ser Ala

35 40 45

Gln Val Ser Gln Ser Asn Pro Ile Gly Phe Tyr His Phe Ala Trp Phe

50 55 60

Gly Gly Asn Glu Glu Glu Ala Glu Ala Glu Ala Arg Tyr Phe Leu Asp

65 70 75 80

Asn Val Pro Thr Gln Val Lys Tyr Leu Val Leu Asp Tyr Glu Asp His

85 90 95

Ala Ser Ala Ser Val Gln Arg Asn Thr Thr Ala Cys Leu Arg Phe Met

100 105 110

Gln Ile Ile Ala Glu Ala Gly Tyr Thr Pro Ile Tyr Tyr Ser Tyr Lys

115 120 125

Pro Phe Thr Leu Asp Asn Val Asp Tyr Gln Gln Ile Leu Ala Gln Phe

130 135 140

Pro Asn Ser Leu Trp Ile Ala Gly Tyr Gly Leu Asn Asp Gly Thr Ala

145 150 155 160

Asn Phe Glu Tyr Phe Pro Ser Met Asp Gly Ile Arg Trp Trp Gln Tyr

165 170 175

Ser Ser Asn Pro Phe Asp Lys Asn Ile Val Leu Leu Asp Asp Glu Lys

180 185 190

Glu Asp Asn Ile Asn Asn Glu Asn Thr Leu Lys Ser Leu Thr Thr Val

195 200 205

Ala Asn Glu Val Ile Gln Gly Leu Trp Gly Asn Gly Gln Glu Arg Tyr

210 215 220

Asp Ser Leu Ala Asn Ala Gly Tyr Asp Pro Gln Ala Val Gln Asp Lys

225 230 235 240

Val Asn Glu Ile Leu Asn Ala Arg Glu Ile Ala Asp Leu Thr Thr Val

245 250 255

Ala Asn Glu Val Ile Gln Gly Leu Trp Gly Asn Gly Gln Glu Arg Tyr

260 265 270

Asp Ser Leu Ala Asn Ala Gly Tyr Asp Pro Gln Ala Val Gln Asp Lys

275 280 285

Val Asn Glu Ile Leu Asn Ala Arg Glu Ile Ala Asp Leu Thr Thr Val

290 295 300

Ala Asn Glu Val Ile Gln Gly Leu Trp Gly Asn Gly Gln Glu Arg Tyr

305 310 315 320

Asp Ser Leu Ala Asn Ala Gly Tyr Asp Pro Gln Ala Val Gln Asp Lys

325 330 335

Val Asn Glu Leu Leu Ser

340

<210> 80

<211> 328

<212> PROTEIN

<213> (Phi)EFAP-1

<400> 80

Met Lys Leu Lys Gly Ile Leu Leu Ser Val Val Thr Thr Phe Gly Leu

1 5 10 15

Leu Phe Gly Ala Thr Asn Val Gln Ala Tyr Glu Val Asn Asn Glu Phe

20 25 30

Asn Leu Gln Pro Trp Glu Gly Ser Gln Gln Leu Ala Tyr Pro Asn Lys

35 40 45

Ile Ile Leu His Glu Thr Ala Asn Pro Arg Ala Thr Gly Arg Asn Glu

50 55 60

Ala Thr Tyr Met Lys Asn Asn Trp Phe Asn Ala His Thr Thr Ala Ile

65 70 75 80

Val Gly Asp Gly Gly Ile Val Tyr Lys Val Ala Pro Glu Gly Asn Val

85 90 95

Ser Trp Gly Ala Gly Asn Ala Asn Pro Tyr Ala Pro Val Gln Ile Glu

100 105 110

Leu Gln His Thr Asn Asp Pro Glu Leu Phe Lys Ala Asn Tyr Lys Ala

115 120 125

Tyr Val Asp Tyr Thr Arg Asp Met Gly Lys Lys Phe Gly Ile Pro Met

130 135 140

Thr Leu Asp Gln Gly Gly Ser Leu Trp Glu Lys Gly Val Val Ser His

145 150 155 160

Gln Trp Val Thr Asp Phe Val Trp Gly Asp His Thr Asp Pro Tyr Gly

165 170 175

Tyr Leu Ala Lys Met Gly Ile Ser Lys Ala Gln Leu Ala His Asp Leu

180 185 190

Ala Asn Gly Val Ser Gly Asn Thr Ala Thr Pro Thr Pro Lys Pro Asp

195 200 205

Lys Pro Lys Pro Thr Gln Pro Ser Lys Pro Ser Asn Lys Lys Arg Phe

210 215 220

Asn Tyr Arg Val Asp Gly Leu Glu Tyr Val Asn Gly Met Trp Gln Ile

225 230 235 240

Tyr Asn Glu His Leu Gly Lys Ile Asp Phe Asn Trp Thr Glu Asn Gly

245 250 255

Ile Pro Val Glu Val Val Asp Lys Val Asn Pro Ala Thr Gly Gln Pro

260 265 270

Thr Lys Asp Gln Val Leu Lys Val Gly Asp Tyr Phe Asn Phe Gln Glu

275 280 285

Asn Ser Thr Gly Val Val Gln Glu Gln Thr Pro Tyr Met Gly Tyr Thr

290 295 300

Leu Ser His Val Gln Leu Pro Asn Glu Phe Ile Trp Leu Phe Thr Asp

305 310 315 320

Ser Lys Gln Ala Leu Met Tyr Gln

325

<210> 81

<211> 48

<212> PROTEIN

<213> Homo sapiens

<400> 81

Ser Ser Leu Leu Glu Lys Gly Leu Asp Gly Ala Lys Lys Ala Val Gly

1 5 10 15

Gly Leu Gly Lys Leu Gly Lys Asp Ala Val Glu Asp Leu Glu Ser Val

20 25 30

Gly Lys Gly Ala Val His Asp Val Lys Asp Val Leu Asp Ser Val Leu

35 40 45

<210> 82

<211> 37

<212> PROTEIN

<213> Hyalophora cecropia

<400> 82

Lys Trp Lys Leu Phe Lys Lys Ile Glu Lys Val Gly Gln Asn Ile Arg

1 5 10 15

Asp Gly Ile Ile Lys Ala Gly Pro Ala Val Ala Val Val Gly Gln Ala

20 25 30

Thr Gln Ile Ala Lys

35

<210> 83

<211> 62

<212> PROTEIN

<213> Drosophila teissieri

<400> 83

Met Lys Tyr Phe Ser Val Leu Val Val Leu Thr Leu Ile Leu Ala Ile

1 5 10 15

Val Asp Gln Ser Asp Ala Phe Ile Asn Leu Leu Asp Lys Val Glu Asp

20 25 30

Ala Leu His Thr Gly Ala Gln Ala Gly Phe Lys Leu Ile Arg Pro Val

35 40 45

Glu Arg Gly Ala Thr Pro Lys Lys Ser Glu Lys Pro Glu Lys

50 55 60

<210> 84

<211> 66

<212> PROTEIN

<213> Bombyx mori

<400> 84

Met Asn Ile Leu Lys Phe Phe Phe Val Phe Ile Val Ala Met Ser Leu

1 5 10 15

Val Ser Cys Ser Thr Ala Ala Pro Ala Lys Ile Pro Ile Lys Ala Ile

20 25 30

Lys Thr Val Gly Lys Ala Val Gly Lys Gly Leu Arg Ala Ile Asn Ile

35 40 45

Ala Ser Thr Ala Asn Asp Val Phe Asn Phe Leu Lys Pro Lys Lys Arg

50 55 60

Lys His

65

<210> 85

<211> 71

<212> PROTEIN

<213> Ceratitis capitata

<400> 85

Met Ala Asn Leu Lys Ala Val Phe Leu Ile Cys Ile Val Ala Phe Ile

1 5 10 15

Ala Leu Gln Cys Val Val Ala Glu Pro Ala Ala Glu Asp Ser Val Val

20 25 30

Val Lys Arg Ser Ile Gly Ser Ala Leu Lys Lys Ala Leu Pro Val Ala

35 40 45

Lys Lys Ile Gly Lys Ile Ala Leu Pro Ile Ala Lys Ala Ala Leu Pro

50 55 60

Val Ala Ala Gly Leu Val Gly

65 70

<210> 86

<211> 53

<212> PROTEIN

<213> Apis mellifera

<400> 86

Met Lys Val Val Ile Phe Ile Phe Ala Leu Leu Ala Thr Ile Cys Ala

1 5 10 15

Ala Phe Ala Tyr Val Pro Leu Pro Asn Val Pro Gln Pro Gly Arg Arg

20 25 30

Pro Phe Pro Thr Phe Pro Gly Gln Gly Pro Phe Asn Pro Lys Ile Lys

35 40 45

Trp Pro Gln Gly Tyr

fifty

<210> 87

<211> 283

<212> PROTEIN

<213> Apis mellifera

<400> 87

Lys Asn Phe Ala Leu Ala Ile Leu Val Val Thr Phe Val Val Ala Val

1 5 10 15

Phe Gly Asn Thr Asn Leu Asp Pro Thr Arg Pro Thr Arg Leu Arg

20 25 30

Arg Glu Ala Lys Pro Glu Ala Glu Pro Gly Asn Asn Arg Pro Val Tyr

35 40 45

Ile Pro Gln Pro Arg Pro Pro His Pro Arg Leu Arg Arg Glu Ala Glu

50 55 60

Pro Glu Ala Glu Pro Gly Asn Asn Arg Pro Val Tyr Ile Pro Gln Pro

65 70 75 80

Arg Pro Pro His Pro Arg Leu Arg Arg Glu Ala Glu Leu Glu Ala Glu

85 90 95

Pro Gly Asn Asn Arg Pro Val Tyr Ile Ser Gln Pro Arg Pro Pro His

100 105 110

Pro Arg Leu Arg Arg Glu Ala Glu Pro Glu Ala Glu Pro Gly Asn Asn

115 120 125

Arg Pro Val Tyr Ile Pro Gln Pro Arg Pro Pro His Pro Arg Leu Arg

130 135 140

Arg Glu Ala Glu Leu Glu Ala Glu Pro Gly Asn Asn Arg Pro Val Tyr

145 150 155 160

Ile Ser Gln Pro Arg Pro Pro His Pro Arg Leu Arg Arg Glu Ala Glu

165 170 175

Pro Glu Ala Glu Pro Gly Asn Asn Arg Pro Val Tyr Ile Pro Gln Pro

180 185 190

Arg Pro Pro His Pro Arg Leu Arg Arg Glu Ala Glu Pro Glu Ala Glu

195 200 205

Pro Gly Asn Asn Arg Pro Val Tyr Ile Pro Gln Pro Arg Pro Pro His

210 215 220

Pro Arg Leu Arg Arg Glu Ala Glu Pro Glu Ala Glu Pro Gly Asn Asn

225 230 235 240

Arg Pro Val Tyr Ile Pro Gln Pro Arg Pro Pro His Pro Arg Leu Arg

245 250 255

Arg Glu Ala Lys Pro Glu Ala Lys Pro Gly Asn Asn Arg Pro Val Tyr

260 265 270

Ile Pro Gln Pro Arg Pro Pro His Pro Arg Ile

275 280

<210> 88

<211> 228

<212> PROTEIN

<213> Sus scrofa

<400> 88

Met Glu Thr Gln Arg Ala Ser Leu Cys Leu Gly Arg Trp Ser Leu Trp

1 5 10 15

Leu Leu Leu Leu Ala Leu Val Val Pro Ser Ala Ser Ala Gln Ala Leu

20 25 30

Ser Tyr Arg Glu Ala Val Leu Arg Ala Val Asp Arg Leu Asn Glu Gln

35 40 45

Ser Ser Glu Ala Asn Leu Tyr Arg Leu Leu Glu Leu Asp Gln Pro Pro

50 55 60

Lys Ala Asp Glu Asp Pro Gly Thr Pro Lys Pro Val Ser Phe Thr Val

65 70 75 80

Lys Glu Thr Val Cys Pro Arg Pro Thr Arg Arg Pro Pro Glu Leu Cys

85 90 95

Asp Phe Lys Glu Asn Gly Arg Val Lys Gln Cys Val Gly Thr Val Thr

100 105 110

Leu Asp Gln Ile Lys Asp Pro Leu Asp Ile Thr Cys Asn Glu Gly Val

115 120 125

Arg Arg Phe Pro Trp Trp Trp Pro Phe Leu Arg Arg Pro Arg Leu Arg

130 135 140

Arg Gln Ala Phe Pro Pro Pro Asn Val Pro Gly Pro Arg Phe Pro Pro

145 150 155 160

Pro Asn Val Pro Gly Pro Arg Phe Pro Pro Pro Asn Phe Pro Gly Pro

165 170 175

Arg Phe Pro Pro Pro Asn Phe Pro Gly Pro Arg Phe Pro Pro Pro Asn

180 185 190

Phe Pro Gly Pro Pro Phe Pro Pro Pro Ile Phe Pro Gly Pro Trp Phe

195 200 205

Pro Pro Pro Pro Pro Phe Arg Pro Pro Pro Phe Gly Pro Pro Arg Phe

210 215 220

Pro Gly Arg Arg

225

<210> 89

<211> 144

<212> PROTEIN

<213> Bos taurus

<400> 89

Met Gln Thr Gln Arg Ala Ser Leu Ser Leu Gly Arg Trp Ser Leu Trp

1 5 10 15

Leu Leu Leu Leu Gly Leu Val Val Pro Ser Ala Ser Ala Gln Ala Leu

20 25 30

Ser Tyr Arg Glu Ala Val Leu Arg Ala Val Asp Gln Leu Asn Glu Leu

35 40 45

Ser Ser Glu Ala Asn Leu Tyr Arg Leu Leu Glu Leu Asp Pro Pro Pro

50 55 60

Lys Asp Asn Glu Asp Leu Gly Thr Arg Lys Pro Val Ser Phe Thr Val

65 70 75 80

Lys Glu Thr Val Cys Pro Arg Thr Ile Gln Gln Pro Ala Glu Gln Cys

85 90 95

Asp Phe Lys Glu Lys Gly Arg Val Lys Gln Cys Val Gly Thr Val Thr

100 105 110

Leu Asp Pro Ser Asn Asp Gln Phe Asp Leu Asn Cys Asn Glu Leu Gln

115 120 125

Ser Val Ile Leu Pro Trp Lys Trp Pro Trp Trp Pro Trp Arg Arg Gly

130 135 140

<210> 90

<211> 149

<212> PROTEIN

<213> Sus scrofa

<400> 90

Met Glu Thr Gln Arg Ala Ser Leu Cys Leu Gly Arg Trp Ser Leu Trp

1 5 10 15

Leu Leu Leu Leu Ala Leu Val Val Pro Ser Ala Ser Ala Gln Ala Leu

20 25 30

Ser Tyr Arg Glu Ala Val Leu Arg Ala Val Asp Arg Leu Asn Glu Gln

35 40 45

Ser Ser Glu Ala Asn Leu Tyr Arg Leu Leu Glu Leu Asp Gln Pro Pro

50 55 60

Lys Ala Asp Glu Asp Pro Gly Thr Pro Lys Pro Val Ser Phe Thr Val

65 70 75 80

Lys Glu Thr Val Cys Pro Arg Pro Thr Arg Gln Pro Pro Glu Leu Cys

85 90 95

Asp Phe Lys Glu Asn Gly Arg Val Lys Gln Cys Val Gly Thr Val Thr

100 105 110

Leu Asp Gln Ile Lys Asp Pro Leu Asp Ile Thr Cys Asn Glu Val Gln

115 120 125

Gly Val Arg Gly Gly Arg Leu Cys Tyr Cys Arg Arg Arg Phe Cys Val

130 135 140

Cys Val Gly Arg Gly

145

<210> 91

<211> 17

<212> PROTEIN

<213> Tachypleus gigas

<400> 91

Lys Trp Cys Phe Arg Val Cys Tyr Arg Gly Ile Cys Tyr Arg Arg Cys

1 5 10 15

Arg

<210> 92

<211> 102

<212> PROTEIN

<213> Anopheles gambiae

<400> 92

Met Lys Cys Ala Thr Ile Val Cys Thr Ile Ala Val Val Leu Ala Ala

1 5 10 15

Thr Leu Leu Asn Gly Ser Val Gln Ala Ala Pro Gln Glu Glu Glu Ala Ala

20 25 30

Leu Ser Gly Gly Ala Asn Leu Asn Thr Leu Leu Asp Glu Leu Pro Glu

35 40 45

Glu Thr His His Ala Ala Leu Glu Asn Tyr Arg Ala Lys Arg Ala Thr

50 55 60

Cys Asp Leu Ala Ser Gly Phe Gly Val Gly Ser Ser Leu Cys Ala Ala

65 70 75 80

His Cys Ile Ala Arg Arg Tyr Arg Gly Gly Tyr Cys Asn Ser Lys Ala

85 90 95

Val Cys Val Cys Arg Asn

100

<210> 93

<211> 70

<212> PROTEIN

<213> Drosophila melanogaster

<400> 93

Met Met Gln Ile Lys Tyr Leu Phe Ala Leu Phe Ala Val Leu Met Leu

1 5 10 15

Val Val Leu Gly Ala Asn Glu Ala Asp Ala Asp Cys Leu Ser Gly Arg

20 25 30

Tyr Lys Gly Pro Cys Ala Val Trp Asp Asn Glu Thr Cys Arg Arg Val

35 40 45

Cys Lys Glu Glu Gly Arg Ser Ser Gly His Cys Ser Pro Ser Leu Lys

50 55 60

Cys Trp Cys Glu Gly Cys

65 70

<210> 94

<211> 63

<212> PROTEIN

<213> Medicago truncatula

<400> 94

Met Thr Lys Ile Val Val Phe Ile Tyr Val Val Ile Leu Leu Leu Thr

1 5 10 15

Ile Phe His Val Ser Ala Lys Lys Lys Arg Tyr Ile Glu Cys Glu Thr

20 25 30

His Glu Asp Cys Ser Gln Val Phe Met Pro Pro Phe Val Met Arg Cys

35 40 45

Val Ile His Glu Cys Lys Ile Phe Asn Gly Glu His Leu Arg Tyr

50 55 60

<210> 95

<211> 76

<212> PROTEIN

<213> Medicago truncatula

<400> 95

Met Ala Lys Ile Met Lys Phe Val Tyr Asn Met Ile Pro Phe Leu Ser

1 5 10 15

Ile Phe Ile Ile Thr Leu Gln Val Asn Val Val Val Cys Glu Ile Asp

20 25 30

Ala Asp Cys Pro Gln Ile Cys Met Pro Pro Tyr Glu Val Arg Cys Val

35 40 45

Asn His Arg Cys Gly Trp Val Asn Thr Asp Asp Ser Leu Phe Leu Thr

50 55 60

Gln Glu Phe Thr Arg Ser Lys Gln Tyr Ile Ile Ser

65 70 75

<210> 96

<211> 76

<212> PROTEIN

<213> Medicago truncatula

<400> 96

Met Tyr Lys Val Val Glu Ser Ile Phe Ile Arg Tyr Met His Arg Lys

1 5 10 15

Pro Asn Met Thr Lys Phe Phe Lys Phe Val Tyr Thr Met Phe Ile Leu

20 25 30

Ile Ser Leu Phe Leu Val Val Thr Asn Ala Asn Ala His Asn Cys Thr

35 40 45

Asp Ile Ser Asp Cys Ser Ser Asn His Cys Ser Tyr Glu Gly Val Ser

50 55 60

Leu Cys Met Asn Gly Gln Cys Ile Cys Ile Tyr Glu

65 70 75

<210> 97

<211> 64

<212> PROTEIN

<213> Medicago truncatula

<400> 97

Met Val Glu Thr Leu Arg Leu Phe Tyr Ile Met Ile Leu Phe Val Ser

1 5 10 15

Leu Cys Leu Val Val Val Asp Gly Glu Ser Lys Leu Glu Gln Thr Cys

20 25 30

Ser Glu Asp Phe Glu Cys Tyr Ile Lys Asn Pro His Val Pro Phe Gly

35 40 45

His Leu Arg Cys Phe Glu Gly Phe Cys Gln Gln Leu Asn Gly Pro Ala

50 55 60

<210> 98

<211> 67

<212> PROTEIN

<213> Medicago truncatula

<400> 98

Met Ala Lys Ile Val Asn Phe Val Tyr Ser Met Ile Val Phe Leu Phe

1 5 10 15

Leu Phe Leu Val Ala Thr Lys Ala Ala Arg Gly Tyr Leu Cys Val Thr

20 25 30

Asp Ser His Cys Pro Pro His Met Cys Pro Pro Gly Met Glu Pro Arg

35 40 45

Cys Val Arg Arg Met Cys Lys Cys Leu Pro Ile Gly Trp Arg Lys Tyr

50 55 60

Phe Val Pro

65

<210> 99

<211> 96

<212> PROTEIN

<213> Medicago truncatula

<400> 99

Met Gln Ile Gly Lys Asn Met Val Glu Thr Pro Lys Leu Asp Tyr Val

1 5 10 15

Ile Ile Phe Phe Phe Leu Tyr Phe Phe Phe Arg Gln Met Ile Ile Leu

20 25 30

Arg Leu Asn Thr Thr Phe Arg Pro Leu Asn Phe Lys Met Leu Arg Phe

35 40 45

Trp Gly Gln Asn Arg Asn Ile Met Lys His Arg Gly Gln Lys Val His

50 55 60

Phe Ser Leu Ile Leu Ser Asp Cys Lys Thr Asn Lys Asp Cys Pro Lys

65 70 75 80

Leu Arg Arg Ala Asn Val Arg Cys Arg Lys Ser Tyr Cys Val Pro Ile

85 90 95

<210> 100

<211> 65

<212> PROTEIN

<213> Medicago truncatula

<400> 100

Met Leu Arg Leu Tyr Leu Val Ser Tyr Phe Leu Leu Lys Arg Thr Leu

1 5 10 15

Leu Val Ser Tyr Phe Ser Tyr Phe Ser Thr Tyr Ile Ile Glu Cys Lys

20 25 30

Thr Asp Asn Asp Cys Pro Ile Ser Gln Leu Lys Ile Tyr Ala Trp Lys

35 40 45

Cys Val Lys Asn Gly Cys His Leu Phe Asp Val Ile Pro Met Met Tyr

50 55 60

Glu

65

<210> 101

<211> 79

<212> PROTEIN

<213> Medicago truncatula

<400> 101

Met Ala Glu Ile Leu Lys Phe Val Tyr Ile Val Ile Leu Phe Val Ser

1 5 10 15

Leu Leu Leu Ile Val Val Ala Ser Glu Arg Glu Cys Val Thr Asp Asp

20 25 30

Asp Cys Glu Lys Leu Tyr Pro Thr Asn Glu Tyr Arg Met Met Cys Asp

35 40 45

Ser Gly Tyr Cys Met Asn Leu Leu Asn Gly Lys Ile Ile Tyr Leu Leu

50 55 60

Cys Leu Lys Lys Lys Lys Phe Leu Ile Ile Ile Ser Val Leu Leu

65 70 75

<210> 102

<211> 95

<212> PROTEIN

<213> Medicago truncatula

<400> 102

Met Ala Glu Ile Ile Lys Phe Val Tyr Ile Met Ile Leu Cys Val Ser

1 5 10 15

Leu Leu Leu Ile Glu Val Ala Gly Glu Glu Cys Val Thr Asp Ala Asp

20 25 30

Cys Asp Lys Leu Tyr Pro Asp Ile Arg Lys Pro Leu Met Cys Ser Ile

35 40 45

Gly Glu Cys Tyr Ser Leu Tyr Lys Gly Lys Phe Ser Leu Ser Ile Ile

50 55 60

Ser Lys Thr Ser Phe Ser Leu Met Val Tyr Asn Val Val Thr Leu Val

65 70 75 80

Ile Cys Leu Arg Leu Ala Tyr Ile Ser Leu Leu Leu Lys Phe Leu

85 90 95

<210> 103

<211> 100

<212> PROTEIN

<213> Medicago truncatula

<400> 103

Met Ala Glu Ile Leu Lys Asp Phe Tyr Ala Met Asn Leu Phe Ile Phe

1 5 10 15

Leu Ile Ile Leu Pro Ala Lys Ile Arg Gly Glu Thr Leu Ser Leu Thr

20 25 30

His Pro Lys Cys His His Ile Met Leu Pro Ser Leu Phe Ile Thr Glu

35 40 45

Val Phe Gln Arg Val Thr Asp Asp Gly Cys Pro Lys Pro Val Asn His

50 55 60

Leu Arg Val Val Lys Cys Ile Glu His Ile Cys Glu Tyr Gly Tyr Asn

65 70 75 80

Tyr Arg Pro Asp Phe Ala Ser Gln Ile Pro Glu Ser Thr Lys Met Pro

85 90 95

Arg Lys Arg Glu

100

<210> 104

<211> 78

<212> PROTEIN

<213> Medicago truncatula

<400> 104

Met Val Glu Ile Leu Lys Asn Phe Tyr Ala Met Asn Leu Phe Ile Phe

1 5 10 15

Leu Ile Ile Leu Ala Val Lys Ile Arg Gly Ala His Phe Pro Cys Val

20 25 30

Thr Asp Asp Asp Cys Pro Lys Pro Val Asn Lys Leu Arg Val Ile Lys

35 40 45

Cys Ile Asp His Ile Cys Gln Tyr Ala Arg Asn Leu Pro Asp Phe Ala

50 55 60

Ser Glu Ile Ser Glu Ser Thr Lys Met Pro Cys Lys Gly Glu

65 70 75

<210> 105

<211> 72

<212> PROTEIN

<213> Medicago truncatula

<400> 105

Met Phe His Ala Gln Ala Glu Asn Met Ala Lys Val Ser Asn Phe Val

1 5 10 15

Cys Ile Met Ile Leu Phe Leu Ala Leu Phe Phe Ile Thr Met Asn Asp

20 25 30

Ala Ala Arg Phe Glu Cys Arg Glu Asp Ser His Cys Val Thr Arg Ile

35 40 45

Lys Cys Val Leu Pro Arg Lys Pro Glu Cys Arg Asn Tyr Ala Cys Gly

50 55 60

Cys Tyr Asp Ser Asn Lys Tyr Arg

65 70

<210> 106

<211> 78

<212> PROTEIN

<213> Medicago truncatula

<400> 106

Met Gln Met Arg Gln Asn Met Ala Thr Ile Leu Asn Phe Val Phe Val

1 5 10 15

Ile Ile Leu Phe Ile Ser Leu Leu Leu Val Val Thr Lys Gly Tyr Arg

20 25 30

Glu Pro Phe Ser Ser Phe Thr Glu Gly Pro Thr Cys Lys Glu Asp Ile

35 40 45

Asp Cys Pro Ser Ile Ser Cys Val Asn Pro Gln Val Pro Lys Cys Ile

50 55 60

Met Phe Glu Cys His Cys Lys Tyr Ile Pro Thr Thr Leu Lys

65 70 75

<210> 107

<211> 71

<212> PROTEIN

<213> Medicago truncatula

<400> 107

Met Ala Thr Ile Leu Met Tyr Val Tyr Ile Thr Ile Leu Phe Ile Ser

1 5 10 15

Ile Leu Thr Val Leu Thr Glu Gly Leu Tyr Glu Pro Leu Tyr Asn Phe

20 25 30

Arg Arg Asp Pro Asp Cys Arg Arg Asn Ile Asp Cys Pro Ser Tyr Leu

35 40 45

Cys Val Ala Pro Lys Val Pro Arg Cys Ile Met Phe Glu Cys His Cys

50 55 60

Lys Asp Ile Pro Ser Asp His

65 70

<210> 108

<211> 57

<212> PROTEIN

<213> Medicago truncatula

<400> 108

Met Thr Thr Ser Leu Lys Phe Val Tyr Val Ala Ile Leu Phe Leu Ser

1 5 10 15

Leu Leu Leu Val Val Met Gly Gly Ile Arg Arg Phe Glu Cys Arg Gln

20 25 30

Asp Ser Asp Cys Pro Ser Tyr Phe Cys Glu Lys Leu Thr Val Pro Lys

35 40 45

Cys Phe Trp Ser Lys Cys Tyr Cys Lys

50 55

<210> 109

<211> 57

<212> PROTEIN

<213> Medicago truncatula

<400> 109

Met Thr Thr Ser Leu Lys Phe Val Tyr Val Ala Ile Leu Phe Leu Ser

1 5 10 15

Leu Leu Leu Val Val Met Gly Gly Ile Arg Lys Lys Glu Cys Arg Gln

20 25 30

Asp Ser Asp Cys Pro Ser Tyr Phe Cys Glu Lys Leu Thr Ile Ala Lys

35 40 45

Cys Ile His Ser Thr Cys Leu Cys Lys

50 55

<210> 110

<211> 66

<212> PROTEIN

<213> Medicago truncatula

<400> 110

Met Gln Ile Gly Lys Asn Met Val Glu Thr Pro Lys Leu Val Tyr Phe

1 5 10 15

Ile Ile Leu Phe Leu Ser Ile Phe Leu Cys Ile Thr Val Ser Asn Ser

20 25 30

Ser Phe Ser Gln Ile Phe Asn Ser Ala Cys Lys Thr Asp Lys Asp Cys

35 40 45

Pro Lys Phe Gly Arg Val Asn Val Arg Cys Arg Lys Gly Asn Cys Val

50 55 60

ProIle

65

<210> 111

<211> 57

<212> PROTEIN

<213> Medicago truncatula

<400> 111

Met Thr Ala Ile Leu Lys Lys Phe Ile Asn Ala Val Phe Leu Phe Ile

1 5 10 15

Val Leu Phe Leu Ala Thr Thr Asn Val Glu Asp Phe Val Gly Gly Ser

20 25 30

Asn Asp Glu Cys Val Tyr Pro Asp Val Phe Gln Cys Ile Asn Asn Ile

35 40 45

Cys Lys Cys Val Ser His His Arg Thr

50 55

<210> 112

<211> 74

<212> PROTEIN

<213> Medicago truncatula

<400> 112

Met Gln Lys Arg Lys Asn Met Ala Gln Ile Ile Phe Tyr Val Tyr Ala

1 5 10 15

Leu Ile Ile Leu Phe Ser Pro Phe Leu Ala Ala Arg Leu Val Phe Val

20 25 30

Asn Pro Glu Lys Pro Cys Val Thr Asp Ala Asp Cys Asp Arg Tyr Arg

35 40 45

His Glu Ser Ala Ile Tyr Ser Asp Met Phe Cys Lys Asp Gly Tyr Cys

50 55 60

Phe Ile Asp Tyr His His Asp Pro Tyr Pro

65 70

<210> 113

<211> 76

<212> PROTEIN

<213> Medicago truncatula

<400> 113

Met Gln Met Arg Lys Asn Met Ala Gln Ile Leu Phe Tyr Val Tyr Ala

1 5 10 15

Leu Leu Ile Leu Phe Thr Pro Phe Leu Val Ala Arg Ile Met Val Val

20 25 30

Asn Pro Asn Asn Pro Cys Val Thr Asp Ala Asp Cys Gln Arg Tyr Arg

35 40 45

His Lys Leu Ala Thr Arg Met Ile Cys Asn Gln Gly Phe Cys Leu Met

50 55 60

Asp Phe Thr His Asp Pro Tyr Ala Pro Ser Leu Pro

65 70 75

<210> 114

<211> 64

<212> PROTEIN

<213> Medicago truncatula

<400> 114

Met Asn His Ile Ser Lys Phe Val Tyr Ala Leu Ile Ile Phe Leu Ser

1 5 10 15

Ile Tyr Leu Val Val Leu Asp Gly Leu Pro Ile Ser Cys Lys Asp His

20 25 30

Phe Glu Cys Arg Arg Lys Ile Asn Ile Leu Arg Cys Ile Tyr Arg Gln

35 40 45

Glu Lys Pro Met Cys Ile Asn Ser Ile Cys Thr Cys Val Lys Leu Leu

50 55 60

<210> 115

<211> 67

<212> PROTEIN

<213> Medicago truncatula

<400> 115

Met Gln Arg Glu Lys Asn Met Ala Lys Ile Phe Glu Phe Val Tyr Ala

1 5 10 15

Met Ile Ile Phe Ile Leu Leu Phe Leu Val Glu Lys Asn Val Val Ala

20 25 30

Tyr Leu Lys Phe Glu Cys Lys Thr Asp Asp Asp Cys Gln Lys Ser Leu

35 40 45

Leu Lys Thr Tyr Val Trp Lys Cys Val Lys Asn Glu Cys Tyr Phe Phe

50 55 60

Ala Lys Lys

65

<210> 116

<211> 61

<212> PROTEIN

<213> Medicago truncatula

<400> 116

Met Ala Gly Ile Ile Lys Phe Val His Val Leu Ile Ile Phe Leu Ser

1 5 10 15

Leu Phe His Val Val Lys Asn Asp Asp Gly Ser Phe Cys Phe Lys Asp

20 25 30

Ser Asp Cys Pro Asp Glu Met Cys Pro Ser Pro Leu Lys Glu Met Cys

35 40 45

Tyr Phe Leu Gln Cys Lys Cys Gly Val Asp Thr Ile Ala

50 55 60

<210> 117

<211> 59

<212> PROTEIN

<213> Medicago truncatula

<400> 117

Met Ala Asn Thr His Lys Leu Val Ser Met Ile Leu Phe Ile Phe Leu

1 5 10 15

Phe Leu Ala Ser Asn Asn Val Glu Gly Tyr Val Asn Cys Glu Thr Asp

20 25 30

Ala Asp Cys Pro Pro Ser Thr Arg Val Lys Arg Phe Lys Cys Val Lys

35 40 45

Gly Glu Cys Arg Trp Thr Arg Met Ser Tyr Ala

50 55

<210> 118

<211> 63

<212> PROTEIN

<213> Medicago truncatula

<400> 118

Met Gln Arg Arg Lys Lys Lys Ala Gln Val Val Met Phe Val His Asp

1 5 10 15

Leu Ile Ile Cys Ile Tyr Leu Phe Ile Val Ile Thr Thr Arg Lys Thr

20 25 30

Asp Ile Arg Cys Arg Phe Tyr Tyr Asp Cys Pro Arg Leu Glu Tyr His

35 40 45

Phe Cys Glu Cys Ile Glu Asp Phe Cys Ala Tyr Ile Arg Leu Asn

50 55 60

<210> 119

<211> 57

<212> PROTEIN

<213> Medicago truncatula

<400> 119

Met Ala Lys Val Tyr Met Phe Val Tyr Ala Leu Ile Ile Phe Val Ser

1 5 10 15

Pro Phe Leu Leu Ala Thr Phe Arg Thr Arg Leu Pro Cys Glu Lys Asp

20 25 30

Asp Asp Cys Pro Glu Ala Phe Leu Pro Val Met Lys Cys Val Asn

35 40 45

Arg Phe Cys Gln Tyr Glu Ile Leu Glu

50 55

<210> 120

<211> 77

<212> PROTEIN

<213> Medicago truncatula

<400> 120

Met Ile Lys Gln Phe Ser Val Cys Tyr Ile Gln Met Arg Arg Asn Met

1 5 10 15

Thr Thr Ile Leu Lys Phe Pro Tyr Ile Met Val Ile Cys Leu Leu Leu

20 25 30

Leu His Val Ala Ala Tyr Glu Asp Phe Glu Lys Glu Ile Phe Asp Cys

35 40 45

Lys Lys Asp Gly Asp Cys Asp His Met Cys Val Thr Pro Gly Ile Pro

50 55 60

Lys Cys Thr Gly Tyr Val Cys Phe Cys Phe Glu Asn Leu

65 70 75

<210> 121

<211> 73

<212> PROTEIN

<213> Medicago truncatula

<400> 121

Met Gln Arg Ser Arg Asn Met Thr Thr Ile Phe Lys Phe Ala Tyr Ile

1 5 10 15

Met Ile Ile Cys Val Phe Leu Leu Asn Ile Ala Ala Gln Glu Ile Glu

20 25 30

Asn Gly Ile His Pro Cys Lys Lys Asn Glu Asp Cys Asn His Met Cys

35 40 45

Val Met Pro Gly Leu Pro Trp Cys His Glu Asn Asn Leu Cys Phe Cys

50 55 60

Tyr Glu Asn Ala Tyr Gly Asn Thr Arg

65 70

<210> 122

<211> 85

<212> PROTEIN

<213> Medicago truncatula

<400> 122

Met Thr Ile Ile Ile Lys Phe Val Asn Val Leu Ile Ile Phe Leu Ser

1 5 10 15

Leu Phe His Val Ala Lys Asn Asp Asp Asn Lys Leu Leu Leu Ser Phe

20 25 30

Ile Glu Glu Gly Phe Leu Cys Phe Lys Asp Ser Asp Cys Pro Tyr Asn

35 40 45

Met Cys Pro Ser Pro Leu Lys Glu Met Cys Tyr Phe Ile Lys Cys Val

50 55 60

Cys Gly Val Tyr Gly Pro Ile Arg Glu Arg Arg Leu Tyr Gln Ser His

65 70 75 80

Asn Pro Met Ile Gln

85

<210> 123

<211> 69

<212> PROTEIN

<213> Medicago truncatula

<400> 123

Met Arg Lys Asn Met Thr Lys Ile Leu Met Ile Gly Tyr Ala Leu Met

1 5 10 15

Ile Phe Ile Phe Leu Ser Ile Ala Val Ser Ile Thr Gly Asn Leu Ala

20 25 30

Arg Ala Ser Arg Lys Lys Pro Val Asp Val Ile Pro Cys Ile Tyr Asp

35 40 45

His Asp Cys Pro Arg Lys Leu Tyr Phe Leu Glu Arg Cys Val Gly Arg

50 55 60

Val Cys Lys Tyr Leu

65

<210> 124

<211> 58

<212> PROTEIN

<213> Medicago truncatula

<400> 124

Met Ala His Lys Leu Val Tyr Ala Ile Thr Leu Phe Ile Phe Leu Phe

1 5 10 15

Leu Ile Ala Asn Asn Ile Glu Asp Asp Ile Phe Cys Ile Thr Asp Asn

20 25 30

Asp Cys Pro Pro Asn Thr Leu Val Gln Arg Tyr Arg Cys Ile Asn Gly

35 40 45

Lys Cys Asn Leu Ser Phe Val Ser Tyr Gly

50 55

<210> 125

<211> 61

<212> PROTEIN

<213> Medicago truncatula

<400> 125

Met Asp Glu Thr Leu Lys Phe Val Tyr Ile Leu Ile Leu Phe Val Ser

1 5 10 15

Leu Cys Leu Val Val Ala Asp Gly Val Lys Asn Ile Asn Arg Glu Cys

20 25 30

Thr Gln Thr Ser Asp Cys Tyr Lys Lys Tyr Pro Phe Ile Pro Trp Gly

35 40 45

Lys Val Arg Cys Val Lys Gly Arg Cys Arg Leu Asp Met

50 55 60

<210> 126

<211> 62

<212> PROTEIN

<213> Medicago truncatula

<400> 126

Met Ala Lys Ile Ile Lys Phe Val Tyr Val Leu Ala Ile Phe Phe Ser

1 5 10 15

Leu Phe Leu Val Ala Lys Asn Val Asn Gly Trp Thr Cys Val Glu Asp

20 25 30

Ser Asp Cys Pro Ala Asn Ile Cys Gln Pro Pro Met Gln Arg Met Cys

35 40 45

Phe Tyr Gly Glu Cys Ala Cys Val Arg Ser Lys Phe Cys Thr

50 55 60

<210> 127

<211> 61

<212> PROTEIN

<213> Medicago truncatula

<400> 127

Met Val Lys Ile Ile Lys Phe Val Tyr Phe Met Thr Leu Phe Leu Ser

1 5 10 15

Met Leu Leu Val Thr Thr Lys Glu Asp Gly Ser Val Glu Cys Ile Ala

20 25 30

Asn Ile Asp Cys Pro Gln Ile Phe Met Leu Pro Phe Val Met Arg Cys

35 40 45

Ile Asn Phe Arg Cys Gln Ile Val Asn Ser Glu Asp Thr

50 55 60

<210> 128

<211> 67

<212> PROTEIN

<213> Medicago truncatula

<400> 128

Met Asp Glu Ile Leu Lys Phe Val Tyr Thr Leu Ile Ile Phe Phe Ser

1 5 10 15

Leu Phe Phe Ala Ala Asn Asn Val Asp Ala Asn Ile Met Asn Cys Gln

20 25 30

Ser Thr Phe Asp Cys Pro Arg Asp Met Cys Ser His Ile Arg Asp Val

35 40 45

Ile Cys Ile Phe Lys Lys Cys Lys Cys Ala Gly Gly Arg Tyr Met Pro

50 55 60

Gln Val Pro

65

<210> 129

<211> 62

<212> PROTEIN

<213> Medicago truncatula

<400> 129

Met Gln Arg Arg Lys Asn Met Ala Asn Asn His Met Leu Ile Tyr Ala

1 5 10 15

Met Ile Ile Cys Leu Phe Pro Tyr Leu Val Val Thr Phe Lys Thr Ala

20 25 30

Ile Thr Cys Asp Cys Asn Glu Asp Cys Leu Asn Phe Phe Thr Pro Leu

35 40 45

Asp Asn Leu Lys Cys Ile Asp Asn Val Cys Glu Val Phe Met

50 55 60

<210> 130

<211> 65

<212> PROTEIN

<213> Medicago truncatula

<400> 130

Met Val Asn Ile Leu Lys Phe Ile Tyr Val Ile Ile Phe Phe Ile Leu

1 5 10 15

Met Phe Phe Val Leu Ile Asp Val Asp Gly His Val Leu Val Glu Cys

20 25 30

Ile Glu Asn Arg Asp Cys Glu Lys Gly Met Cys Lys Phe Pro Phe Ile

35 40 45

Val Arg Cys Leu Met Asp Gln Cys Lys Cys Val Arg Ile His Asn Leu

50 55 60

ile

65

<210> 131

<211> 74

<212> PROTEIN

<213> Medicago truncatula

<400> 131

Met Ile Ile Gln Phe Ser Ile Tyr Tyr Met Gln Arg Arg Lys Leu Asn

1 5 10 15

Met Val Glu Ile Leu Lys Phe Ser His Ala Leu Ile Ile Phe Leu Phe

20 25 30

Leu Ser Ala Leu Val Thr Asn Ala Asn Ile Phe Phe Cys Ser Thr Asp

35 40 45

Glu Asp Cys Thr Trp Asn Leu Cys Arg Gln Pro Trp Val Gln Lys Cys

50 55 60

Arg Leu His Met Cys Ser Cys Glu Lys Asn

65 70

<210> 132

<211> 58

<212> PROTEIN

<213> Medicago truncatula

<400> 132

Met Asp Glu Val Phe Lys Phe Val Tyr Val Met Ile Ile Phe Pro Phe

1 5 10 15

Leu Ile Leu Asp Val Ala Thr Asn Ala Glu Lys Ile Arg Arg Cys Phe

20 25 30

Asn Asp Ala His Cys Pro Pro Asp Met Cys Thr Leu Gly Val Ile Pro

35 40 45

Lys Cys Ser Arg Phe Thr Ile Cys Ile Cys

50 55

<210> 133

<211> 64

<212> PROTEIN

<213> Medicago truncatula

<400> 133

Met His Arg Lys Pro Asn Met Thr Lys Phe Phe Lys Phe Val Tyr Thr

1 5 10 15

Met Phe Ile Leu Ile Ser Leu Phe Leu Val Val Thr Asn Ala Asn Ala

20 25 30

Asn Asn Cys Thr Asp Thr Ser Asp Cys Ser Ser Asn His Cys Ser Tyr

35 40 45

Glu Gly Val Ser Leu Cys Met Asn Gly Gln Cys Ile Cys Ile Tyr Glu

50 55 60

<210> 134

<211> 69

<212> PROTEIN

<213> Medicago truncatula

<400> 134

Met Gln Met Lys Lys Met Ala Thr Ile Leu Lys Phe Val Tyr Leu Ile

1 5 10 15

Ile Leu Leu Ile Tyr Pro Leu Leu Val Val Thr Glu Glu Ser His Tyr

20 25 30

Met Lys Phe Ser Ile Cys Lys Asp Asp Thr Asp Cys Pro Thr Leu Phe

35 40 45

Cys Val Leu Pro Asn Val Pro Lys Cys Ile Gly Ser Lys Cys His Cys

50 55 60

Lys Leu Met Val Asn

65

<210> 135

<211> 64

<212> PROTEIN

<213> Medicago truncatula

<400> 135

Met Val Glu Thr Leu Arg Leu Phe Tyr Ile Met Ile Leu Phe Val Ser

1 5 10 15

Leu Tyr Leu Val Val Val Asp Gly Val Ser Lys Leu Ala Gln Ser Cys

20 25 30

Ser Glu Asp Phe Glu Cys Tyr Ile Lys Asn Pro His Ala Pro Phe Gly

35 40 45

Gln Leu Arg Cys Phe Glu Gly Tyr Cys Gln Arg Leu Asp Lys Pro Thr

50 55 60

<210> 136

<211> 63

<212> PROTEIN

<213> Medicago truncatula

<400> 136

Met Thr Thr Phe Leu Lys Val Ala Tyr Ile Met Ile Ile Cys Val Phe

1 5 10 15

Val Leu His Leu Ala Ala Gln Val Asp Ser Gln Lys Arg Leu His Gly

20 25 30

Cys Lys Glu Asp Arg Asp Cys Asp Asn Ile Cys Ser Val His Ala Val

35 40 45

Thr Lys Cys Ile Gly Asn Met Cys Arg Cys Leu Ala Asn Val Lys

50 55 60

<210> 137

<211> 61

<212> PROTEIN

<213> Medicago truncatula

<400> 137

Met Arg Ile Asn Arg Thr Pro Ala Ile Phe Lys Phe Val Tyr Thr Ile

1 5 10 15

Ile Ile Tyr Leu Phe Leu Leu Arg Val Val Ala Lys Asp Leu Pro Phe

20 25 30

Asn Ile Cys Glu Lys Asp Glu Asp Cys Leu Glu Phe Cys Ala His Asp

35 40 45

Lys Val Ala Lys Cys Met Leu Asn Ile Cys Phe Cys Phe

50 55 60

<210> 138

<211> 54

<212> PROTEIN

<213> Medicago truncatula

<400> 138

Met Ala Glu Ile Leu Lys Ile Leu Tyr Val Phe Ile Ile Phe Leu Ser

1 5 10 15

Leu Ile Leu Ala Val Ile Ser Gln His Pro Phe Thr Pro Cys Glu Thr

20 25 30

Asn Ala Asp Cys Lys Cys Arg Asn His Lys Arg Pro Asp Cys Leu Trp

35 40 45

His Lys Cys Tyr Cys Tyr

fifty

<210> 139

<211> 65

<212> PROTEIN

<213> Medicago truncatula

<400> 139

Met Arg Lys Ser Met Ala Thr Ile Leu Lys Phe Val Tyr Val Ile Met

1 5 10 15

Leu Phe Ile Tyr Ser Leu Phe Val Ile Glu Ser Phe Gly His Arg Phe

20 25 30

Leu Ile Tyr Asn Asn Cys Lys Asn Asp Thr Glu Cys Pro Asn Asp Cys

35 40 45

Gly Pro His Glu Gln Ala Lys Cys Ile Leu Tyr Ala Cys Tyr Cys Val

50 55 60

Glu

65

<210> 140

<211> 58

<212> PROTEIN

<213> Medicago truncatula

<400> 140

Met Asn Thr Ile Leu Lys Phe Ile Phe Val Val Phe Leu Phe Leu Ser

1 5 10 15

Ile Phe Leu Ser Ala Gly Asn Ser Lys Ser Tyr Gly Pro Cys Thr Thr

20 25 30

Leu Gln Asp Cys Glu Thr His Asn Trp Phe Glu Val Cys Ser Cys Ile

35 40 45

Asp Phe Glu Cys Lys Cys Trp Ser Leu Leu

50 55

<210> 141

<211> 64

<212> PROTEIN

<213> Medicago truncatula

<400> 141

Met Ala Glu Ile Ile Lys Phe Val Tyr Ile Met Ile Leu Cys Val Ser

1 5 10 15

Leu Leu Leu Ile Ala Glu Ala Ser Gly Lys Glu Cys Val Thr Asp Ala

20 25 30

Asp Cys Glu Asn Leu Tyr Pro Gly Asn Lys Lys Pro Met Phe Cys Asn

35 40 45

Asn Thr Gly Tyr Cys Met Ser Leu Tyr Lys Glu Pro Ser Arg Tyr Met

50 55 60

<210> 142

<211> 64

<212> PROTEIN

<213> Medicago truncatula

<400> 142

Met Ala Lys Ile Ile Lys Phe Val Tyr Ile Met Ile Leu Cys Val Ser

1 5 10 15

Leu Leu Leu Ile Val Glu Ala Gly Gly Lys Glu Cys Val Thr Asp Val

20 25 30

Asp Cys Glu Lys Ile Tyr Pro Gly Asn Lys Lys Pro Leu Ile Cys Ser

35 40 45

Thr Gly Tyr Cys Tyr Ser Leu Tyr Glu Pro Pro Arg Tyr His Lys

50 55 60

<210> 143

<211> 64

<212> PROTEIN

<213> Medicago truncatula

<400> 143

Met Ala Lys Val Thr Lys Phe Gly Tyr Ile Ile Ile His Phe Leu Ser

1 5 10 15

Leu Phe Phe Leu Ala Met Asn Val Ala Gly Gly Arg Glu Cys His Ala

20 25 30

Asn Ser His Cys Val Gly Lys Ile Thr Cys Val Leu Pro Gln Lys Pro

35 40 45

Glu Cys Trp Asn Tyr Ala Cys Val Cys Tyr Asp Ser Asn Lys Tyr Arg

50 55 60

<210> 144

<211> 55

<212> PROTEIN

<213> Medicago truncatula

<400> 144

Met Ala Lys Ile Phe Asn Tyr Val Tyr Ala Leu Ile Met Phe Leu Ser

1 5 10 15

Leu Phe Leu Met Gly Thr Ser Gly Met Lys Asn Gly Cys Lys His Thr

20 25 30

Gly His Cys Pro Arg Lys Met Cys Gly Ala Lys Thr Thr Lys Cys Arg

35 40 45

Asn Asn Lys Cys Gln Cys Val

50 55

<210> 145

<211> 69

<212> PROTEIN

<213> Medicago truncatula

<400> 145

Met Thr Glu Ile Leu Lys Phe Val Cys Val Met Ile Ile Phe Ile Ser

1 5 10 15

Ser Phe Ile Val Ser Lys Ser Leu Asn Gly Gly Gly Lys Asp Lys Cys

20 25 30

Phe Arg Asp Ser Asp Cys Pro Lys His Met Cys Pro Ser Ser Leu Val

35 40 45

Ala Lys Cys Ile Asn Arg Leu Cys Arg Cys Arg Arg Pro Glu Leu Gln

50 55 60

Val Gln Leu Asn Pro

65

<210> 146

<211> 69

<212> PROTEIN

<213> Medicago truncatula

<400> 146

Met Ala His Ile Ile Met Phe Val Tyr Ala Leu Ile Tyr Ala Leu Ile

1 5 10 15

Ile Phe Ser Ser Leu Phe Val Arg Asp Gly Ile Pro Cys Leu Ser Asp

20 25 30

Asp Glu Cys Pro Glu Met Ser His Tyr Ser Phe Lys Cys Asn Asn Lys

35 40 45

Ile Cys Glu Tyr Asp Leu Gly Glu Met Ser Asp Asp Asp Tyr Tyr Leu

50 55 60

Glu Met Ser Arg Glu

65

<210> 147

<211> 77

<212> PROTEIN

<213> Medicago truncatula

<400> 147

Met Tyr Arg Glu Lys Asn Met Ala Lys Thr Leu Lys Phe Val Tyr Val

1 5 10 15

Ile Val Leu Phe Leu Ser Leu Phe Leu Ala Ala Lys Asn Ile Asp Gly

20 25 30

Arg Val Ser Tyr Asn Ser Phe Ile Ala Leu Pro Val Cys Gln Thr Ala

35 40 45

Ala Asp Cys Pro Glu Gly Thr Arg Gly Arg Thr Tyr Lys Cys Ile Asn

50 55 60

Asn Lys Cys Arg Tyr Pro Lys Leu Leu Lys Pro Ile Gln

65 70 75

<210> 148

<211> 56

<212> PROTEIN

<213> Medicago truncatula

<400> 148

Met Ala His Ile Phe Asn Tyr Val Tyr Ala Leu Leu Val Phe Leu Ser

1 5 10 15

Leu Phe Leu Met Val Thr Asn Gly Ile His Ile Gly Cys Asp Lys Asp

20 25 30

Arg Asp Cys Pro Lys Gln Met Cys His Leu Asn Gln Thr Pro Lys Cys

35 40 45

Leu Lys Asn Ile Cys Lys Cys Val

50 55

<210> 149

<211> 77

<212> PROTEIN

<213> Medicago truncatula

<400> 149

Met Ala Glu Ile Leu Lys Cys Phe Tyr Thr Met Asn Leu Phe Ile Phe

1 5 10 15

Leu Ile Ile Leu Pro Ala Lys Ile Arg Glu His Ile Gln Cys Val Ile

20 25 30

Asp Asp Asp Cys Pro Lys Ser Leu Asn Lys Leu Leu Ile Ile Lys Cys

35 40 45

Ile Asn His Val Cys Gln Tyr Val Gly Asn Leu Pro Asp Phe Ala Ser

50 55 60

Gln Ile Pro Lys Ser Thr Lys Met Pro Tyr Lys Gly Glu

65 70 75

<210> 150

<211> 70

<212> PROTEIN

<213> Medicago truncatula

<400> 150

Met Ala Tyr Ile Ser Arg Ile Phe Tyr Val Leu Ile Ile Phe Leu Ser

1 5 10 15

Leu Phe Phe Val Val Ile Asn Gly Val Lys Ser Leu Leu Leu Ile Lys

20 25 30

Val Arg Ser Phe Ile Pro Cys Gln Arg Ser Asp Asp Cys Pro Arg Asn

35 40 45

Leu Cys Val Asp Gln Ile Ile Pro Thr Cys Val Trp Ala Lys Cys Lys

50 55 60

Cys Lys Asn Tyr Asn Asp

65 70

<210> 151

<211> 64

<212> PROTEIN

<213> Medicago truncatula

<400> 151

Met Ala Asn Val Thr Lys Phe Val Tyr Ile Ala Ile Tyr Phe Leu Ser

1 5 10 15

Leu Phe Phe Ile Ala Lys Asn Asp Ala Thr Ala Thr Phe Cys His Asp

20 25 30

Asp Ser His Cys Val Thr Lys Ile Lys Cys Val Leu Pro Arg Thr Pro

35 40 45

Gln Cys Arg Asn Glu Ala Cys Gly Cys Tyr His Ser Asn Lys Phe Arg

50 55 60

<210> 152

<211> 62

<212> PROTEIN

<213> Medicago truncatula

<400> 152

Met Gly Glu Ile Met Lys Phe Val Tyr Val Met Ile Ile Tyr Leu Phe

1 5 10 15

Met Phe Asn Val Ala Thr Gly Ser Glu Phe Ile Phe Thr Lys Lys Leu

20 25 30

Thr Ser Cys Asp Ser Ser Lys Asp Cys Arg Ser Phe Leu Cys Tyr Ser

35 40 45

Pro Lys Phe Pro Val Cys Lys Arg Gly Ile Cys Glu Cys Ile

50 55 60

<210> 153

<211> 61

<212> PROTEIN

<213> Medicago truncatula

<400> 153

Met Gly Glu Met Phe Lys Phe Ile Tyr Thr Phe Ile Leu Phe Val His

1 5 10 15

Leu Phe Leu Val Val Ile Phe Glu Asp Ile Gly His Ile Lys Tyr Cys

20 25 30

Gly Ile Val Asp Asp Cys Tyr Lys Ser Lys Lys Pro Leu Phe Lys Ile

35 40 45

Trp Lys Cys Val Glu Asn Val Cys Val Leu Trp Tyr Lys

50 55 60

<210> 154

<211> 63

<212> PROTEIN

<213> Medicago truncatula

<400> 154

Met Ala Arg Thr Leu Lys Phe Val Tyr Ser Met Ile Leu Phe Leu Ser

1 5 10 15

Leu Phe Leu Val Ala Asn Gly Leu Lys Ile Phe Cys Ile Asp Val Ala

20 25 30

Asp Cys Pro Lys Asp Leu Tyr Pro Leu Leu Tyr Lys Cys Ile Tyr Asn

35 40 45

Lys Cys Ile Val Phe Thr Arg Ile Pro Phe Pro Phe Asp Trp Ile

50 55 60

<210> 155

<211> 66

<212> PROTEIN

<213> Medicago truncatula

<400> 155

Met Ala Asn Ile Thr Lys Phe Val Tyr Ile Ala Ile Leu Phe Leu Ser

1 5 10 15

Leu Phe Phe Ile Gly Met Asn Asp Ala Ala Ile Leu Glu Cys Arg Glu

20 25 30

Asp Ser His Cys Val Thr Lys Ile Lys Cys Val Leu Pro Arg Lys Pro

35 40 45

Glu Cys Arg Asn Asn Ala Cys Thr Cys Tyr Lys Gly Gly Phe Ser Phe

50 55 60

His His

65

<210> 156

<211> 68

<212> PROTEIN

<213> Medicago truncatula

<400> 156

Met Gln Arg Val Lys Lys Met Ser Glu Thr Leu Lys Phe Val Tyr Val

1 5 10 15

Leu Ile Leu Phe Ile Ser Ile Phe His Val Val Ile Val Cys Asp Ser

20 25 30

Ile Tyr Phe Pro Val Ser Arg Pro Cys Ile Thr Asp Lys Asp Cys Pro

35 40 45

Asn Met Lys His Tyr Lys Ala Lys Cys Arg Lys Gly Phe Cys Ile Ser

50 55 60

Ser Arg Val Arg

65

<210> 157

<211> 72

<212> PROTEIN

<213> Medicago truncatula

<400> 157

Met Gln Ile Arg Lys Ile Met Ser Gly Val Leu Lys Phe Val Tyr Ala

1 5 10 15

Ile Ile Leu Phe Leu Phe Leu Phe Leu Val Ala Arg Glu Val Gly Gly

20 25 30

Leu Glu Thr Ile Glu Cys Glu Thr Asp Gly Asp Cys Pro Arg Ser Met

35 40 45

Ile Lys Met Trp Asn Lys Asn Tyr Arg His Lys Cys Ile Asp Gly Lys

50 55 60

Cys Glu Trp Ile Lys Lys Leu Pro

65 70

<210> 158

<211> 54

<212> PROTEIN

<213> Medicago truncatula

<400> 158

Met Phe Val Tyr Asp Leu Ile Leu Phe Ile Ser Leu Ile Leu Val Val

1 5 10 15

Thr Gly Ile Asn Ala Glu Ala Asp Thr Ser Cys His Ser Phe Asp Asp

20 25 30

Cys Pro Trp Val Ala His His Tyr Arg Glu Cys Ile Glu Gly Leu Cys

35 40 45

Ala Tyr Arg Ile Leu Tyr

fifty

<210> 159

<211> 69

<212> PROTEIN

<213> Medicago truncatula

<400> 159

Met Gln Arg Arg Lys Lys Ser Met Ala Lys Met Leu Lys Phe Phe Phe

1 5 10 15

Ala Ile Ile Leu Leu Leu Ser Leu Phe Leu Val Ala Thr Glu Val Gly

20 25 30

Gly Ala Tyr Ile Glu Cys Glu Val Asp Asp Asp Cys Pro Lys Pro Met

35 40 45

Lys Asn Ser His Pro Asp Thr Tyr Tyr Lys Cys Val Lys His Arg Cys

50 55 60

Gln Trp Ala Trp Lys

65

<210> 160

<211> 140

<212> PROTEIN

<213> Medicago truncatula

<400> 160

Met Phe Val Tyr Thr Leu Ile Ile Phe Leu Phe Pro Ser His Val Ile

1 5 10 15

Thr Asn Lys Ile Ala Ile Tyr Cys Val Ser Asp Asp Asp Cys Leu Lys

20 25 30

Thr Phe Thr Pro Leu Asp Leu Lys Cys Val Asp Asn Val Cys Glu Phe

35 40 45

Asn Leu Arg Cys Lys Gly Lys Cys Gly Glu Arg Asp Glu Lys Phe Val

50 55 60

Phe Leu Lys Ala Leu Lys Lys Met Asp Gln Lys Leu Val Leu Glu Glu

65 70 75 80

Gln Gly Asn Ala Arg Glu Val Lys Ile Pro Lys Lys Leu Leu Phe Asp

85 90 95

Arg Ile Gln Val Pro Thr Pro Ala Thr Lys Asp Gln Val Glu Glu Asp

100 105 110

Asp Tyr Asp Asp Asp Asp Glu Glu Glu Glu Glu Glu Glu Asp Asp Val

115 120 125

Asp Met Trp Phe His Leu Pro Asp Val Val Cys His

130 135 140

<210> 161

<211> 60

<212> PROTEIN

<213> Medicago truncatula

<400> 161

Met Ala Lys Phe Ser Met Phe Val Tyr Ala Leu Ile Asn Phe Leu Ser

1 5 10 15

Leu Phe Leu Val Glu Thr Ala Ile Thr Asn Ile Arg Cys Val Ser Asp

20 25 30

Asp Asp Cys Pro Lys Val Ile Lys Pro Leu Val Met Lys Cys Ile Gly

35 40 45

Asn Tyr Cys Tyr Phe Phe Met Ile Tyr Glu Gly Pro

50 55 60

<210> 162

<211> 58

<212> PROTEIN

<213> Medicago truncatula

<400> 162

Met Ala His Lys Phe Val Tyr Ala Ile Ile Leu Phe Ile Phe Leu Phe

1 5 10 15

Leu Val Ala Lys Asn Val Lys Gly Tyr Val Val Cys Arg Thr Val Asp

20 25 30

Asp Cys Pro Pro Asp Thr Arg Asp Leu Arg Tyr Arg Cys Leu Asn Gly

35 40 45

Lys Cys Lys Ser Tyr Arg Leu Ser Tyr Gly

50 55

<210> 163

<211> 61

<212> PROTEIN

<213> Medicago truncatula

<400> 163

Met Gln Arg Lys Lys Asn Met Gly Gln Ile Leu Ile Phe Val Phe Ala

1 5 10 15

Leu Ile Asn Phe Leu Ser Pro Ile Leu Val Glu Met Thr Thr Thr Thr

20 25 30

Ile Pro Cys Thr Phe Ile Asp Asp Cys Pro Lys Met Pro Leu Val Val

35 40 45

Lys Cys Ile Asp Asn Phe Cys Asn Tyr Phe Glu Ile Lys

50 55 60

<210> 164

<211> 57

<212> PROTEIN

<213> Medicago truncatula

<400> 164

Met Ala Gln Thr Leu Met Leu Val Tyr Ala Leu Ile Ile Phe Thr Ser

1 5 10 15

Leu Phe Leu Val Val Ile Ser Arg Gln Thr Asp Ile Pro Cys Lys Ser

20 25 30

Asp Asp Ala Cys Pro Arg Val Ser Ser His His Ile Glu Cys Val Lys

35 40 45

Gly Phe Cys Thr Tyr Trp Lys Leu Asp

50 55

<210> 165

<211> 77

<212> PROTEIN

<213> Medicago truncatula

<400> 165

Met Leu Arg Arg Lys Asn Thr Val Gln Ile Leu Met Phe Val Ser Ala

1 5 10 15

Leu Leu Ile Tyr Ile Phe Leu Phe Leu Val Ile Thr Ser Ser Ala Asn

20 25 30

Ile Pro Cys Asn Ser Asp Ser Asp Cys Pro Trp Lys Ile Tyr Tyr Thr

35 40 45

Tyr Arg Cys Asn Asp Gly Phe Cys Val Tyr Lys Ser Ile Asp Pro Ser

50 55 60

Thr Ile Pro Gln Tyr Met Thr Asp Leu Ile Phe Pro Arg

65 70 75

<210> 166

<211> 59

<212> PROTEIN

<213> Medicago truncatula

<400> 166

Met Ala Val Ile Leu Lys Phe Val Tyr Ile Met Ile Ile Phe Leu Phe

1 5 10 15

Leu Leu Tyr Val Val Asn Gly Thr Arg Cys Asn Arg Asp Glu Asp Cys

20 25 30

Pro Phe Ile Cys Thr Gly Pro Gln Ile Pro Lys Cys Val Ser His Ile

35 40 45

Cys Phe Cys Leu Ser Ser Gly Lys Glu Ala Tyr

50 55

<210> 167

<211> 62

<212> PROTEIN

<213> Medicago truncatula

<400> 167

Met Asp Ala Ile Leu Lys Phe Ile Tyr Ala Met Phe Leu Phe Leu Phe

1 5 10 15

Leu Phe Val Thr Thr Arg Asn Val Glu Ala Leu Phe Glu Cys Asn Arg

20 25 30

Asp Phe Val Cys Gly Asn Asp Asp Glu Cys Val Tyr Pro Tyr Ala Val

35 40 45

Gln Cys Ile His Arg Tyr Cys Lys Cys Leu Lys Ser Arg Asn

50 55 60

<210> 168

<211> 67

<212> PROTEIN

<213> Medicago truncatula

<400> 168

Met Gln Ile Gly Arg Lys Lys Met Gly Glu Thr Pro Lys Leu Val Tyr

1 5 10 15

Val Ile Ile Leu Phe Leu Ser Ile Phe Leu Cys Thr Asn Ser Ser Phe

20 25 30

Ser Gln Met Ile Asn Phe Arg Gly Cys Lys Arg Asp Lys Asp Cys Pro

35 40 45

Gln Phe Arg Gly Val Asn Ile Arg Cys Arg Ser Gly Phe Cys Thr Pro

50 55 60

Ile Asp Ser

65

<210> 169

<211> 76

<212> PROTEIN

<213> Medicago truncatula

<400> 169

Met Gln Met Arg Lys Asn Met Ala Gln Ile Leu Phe Tyr Val Tyr Ala

1 5 10 15

Leu Leu Ile Leu Phe Ser Pro Phe Leu Val Ala Arg Ile Met Val Val

20 25 30

Asn Pro Asn Asn Pro Cys Val Thr Asp Ala Asp Cys Gln Arg Tyr Arg

35 40 45

His Lys Leu Ala Thr Arg Met Val Cys Asn Ile Gly Phe Cys Leu Met

50 55 60

Asp Phe Thr His Asp Pro Tyr Ala Pro Ser Leu Pro

65 70 75

<210> 170

<211> 77

<212> PROTEIN

<213> Medicago truncatula

<400> 170

Met Tyr Val Tyr Tyr Ile Gln Met Gly Lys Asn Met Ala Gln Arg Phe

1 5 10 15

Met Phe Ile Tyr Ala Leu Ile Ile Phe Leu Ser Gln Phe Phe Val Val

20 25 30

Ile Asn Thr Ser Asp Ile Pro Asn Asn Ser Asn Arg Asn Ser Pro Lys

35 40 45

Glu Asp Val Phe Cys Asn Ser Asn Asp Asp Cys Pro Thr Ile Leu Tyr

50 55 60

Tyr Val Ser Lys Cys Val Tyr Asn Phe Cys Glu Tyr Trp

65 70 75

<210> 171

<211> 67

<212> PROTEIN

<213> Medicago truncatula

<400> 171

Met Ala Lys Ile Val Asn Phe Val Tyr Ser Met Ile Ile Phe Val Ser

1 5 10 15

Leu Phe Leu Val Ala Thr Lys Gly Gly Ser Lys Pro Phe Leu Thr Arg

20 25 30

Pro Tyr Pro Cys Asn Thr Gly Ser Asp Cys Pro Gln Asn Met Cys Pro

35 40 45

Pro Gly Tyr Lys Pro Gly Cys Glu Asp Gly Tyr Cys Asn His Cys Tyr

50 55 60

Lys Arg Trp

65

<210> 172

<211> 62

<212> PROTEIN

<213> Medicago truncatula

<400> 172

Met Val Arg Thr Leu Lys Phe Val Tyr Val Ile Ile Leu Ile Leu Ser

1 5 10 15

Leu Phe Leu Val Ala Lys Gly Gly Gly Lys Lys Ile Tyr Cys Glu Asn

20 25 30

Ala Ala Ser Cys Pro Arg Leu Met Tyr Pro Leu Val Tyr Lys Cys Leu

35 40 45

Asp Asn Lys Cys Val Lys Phe Met Met Lys Ser Arg Phe Val

50 55 60

<210> 173

<211> 62

<212> PROTEIN

<213> Medicago truncatula

<400> 173

Met Ala Arg Thr Leu Lys Phe Val Tyr Ala Val Ile Leu Phe Leu Ser

1 5 10 15

Leu Phe Leu Val Ala Lys Gly Asp Asp Val Lys Ile Lys Cys Val Val

20 25 30

Ala Ala Asn Cys Pro Asp Leu Met Tyr Pro Leu Val Tyr Lys Cys Leu

35 40 45

Asn Gly Ile Cys Val Gln Phe Thr Leu Thr Phe Pro Phe Val

50 55 60

<210> 174

<211> 65

<212> PROTEIN

<213> Medicago truncatula

<400> 174

Met Ser Asn Thr Leu Met Phe Val Ile Thr Phe Ile Val Leu Val Thr

1 5 10 15

Leu Phe Leu Gly Pro Lys Asn Val Tyr Ala Phe Gln Pro Cys Val Thr

20 25 30

Thr Ala Asp Cys Met Lys Thr Leu Lys Thr Asp Glu Asn Ile Trp Tyr

35 40 45

Glu Cys Ile Asn Asp Phe Cys Ile Pro Phe Pro Ile Pro Lys Gly Arg

50 55 60

Lys

65

<210> 175

<211> 76

<212> PROTEIN

<213> Medicago truncatula

<400> 175

Met Lys Arg Val Val Asn Met Ala Lys Ile Val Lys Tyr Val Tyr Val

1 5 10 15

Ile Ile Ile Phe Leu Ser Leu Phe Leu Val Ala Thr Lys Ile Glu Gly

20 25 30

Tyr Tyr Tyr Lys Cys Phe Lys Asp Ser Asp Cys Val Lys Leu Leu Cys

35 40 45

Arg Ile Pro Leu Arg Pro Lys Cys Met Tyr Arg His Ile Cys Lys Cys

50 55 60

Lys Val Val Leu Thr Gln Asn Asn Tyr Val Leu Thr

65 70 75

<210> 176

<211> 66

<212> PROTEIN

<213> Medicago truncatula

<400> 176

Met Lys Arg Gly Lys Asn Met Ser Lys Ile Leu Lys Phe Ile Tyr Ala

1 5 10 15

Thr Leu Val Leu Tyr Leu Phe Leu Val Val Thr Lys Ala Ser Asp Asp

20 25 30

Glu Cys Lys Ile Asp Gly Asp Cys Pro Ile Ser Trp Gln Lys Phe His

35 40 45

Thr Tyr Lys Cys Ile Asn Gln Lys Cys Lys Trp Val Leu Arg Phe His

50 55 60

Glu Tyr

65

<210> 177

<211> 64

<212> PROTEIN

<213> Medicago truncatula

<400> 177

Met Ala Lys Thr Leu Asn Phe Met Phe Ala Leu Ile Leu Phe Ile Ser

1 5 10 15

Leu Phe Leu Val Ser Lys Asn Val Ala Ile Asp Ile Phe Val Cys Gln

20 25 30

Thr Asp Ala Asp Cys Pro Lys Ser Glu Leu Ser Met Tyr Thr Trp Lys

35 40 45

Cys Ile Asp Asn Glu Cys Asn Leu Phe Lys Val Met Gln Gln Met Val

50 55 60

<210> 178

<211> 59

<212> PROTEIN

<213> Medicago truncatula

<400> 178

Met Ala Asn Thr His Lys Leu Val Ser Met Ile Leu Phe Ile Phe Leu

1 5 10 15

Phe Leu Val Ala Asn Asn Val Glu Gly Tyr Val Asn Cys Glu Thr Asp

20 25 30

Ala Asp Cys Pro Pro Ser Thr Arg Val Lys Arg Phe Lys Cys Val Lys

35 40 45

Gly Glu Cys Arg Trp Thr Arg Met Ser Tyr Ala

50 55

<210> 179

<211> 59

<212> PROTEIN

<213> Medicago truncatula

<400> 179

Met Ala His Phe Leu Met Phe Val Tyr Ala Leu Ile Thr Cys Leu Ser

1 5 10 15

Leu Phe Leu Val Glu Met Gly His Leu Ser Ile His Cys Val Ser Val

20 25 30

Asp Asp Cys Pro Lys Val Glu Lys Pro Ile Thr Met Lys Cys Ile Asn

35 40 45

Asn Tyr Cys Lys Tyr Phe Val Asp His Lys Leu

50 55

<210> 180

<211> 66

<212> PROTEIN

<213> Medicago truncatula

<400> 180

Met Asn Gln Ile Pro Met Phe Gly Tyr Thr Leu Ile Ile Phe Phe Ser

1 5 10 15

Leu Phe Pro Val Ile Thr Asn Gly Asp Arg Ile Pro Cys Val Thr Asn

20 25 30

Gly Asp Cys Pro Val Met Arg Leu Pro Leu Tyr Met Arg Cys Ile Thr

35 40 45

Tyr Ser Cys Glu Leu Phe Phe Asp Gly Pro Asn Leu Cys Ala Val Glu

50 55 60

Arg Ile

65

<210> 181

<211> 61

<212> PROTEIN

<213> Medicago truncatula

<400> 181

Met Arg Lys Asp Met Ala Arg Ile Ser Leu Phe Val Tyr Ala Leu Ile

1 5 10 15

Ile Phe Phe Ser Leu Phe Phe Val Leu Thr Asn Gly Glu Leu Glu Ile

20 25 30

Arg Cys Val Ser Asp Ala Asp Cys Pro Leu Phe Pro Leu Pro Leu His

35 40 45

Asn Arg Cys Ile Asp Asp Val Cys His Leu Phe Thr Ser

50 55 60

<210> 182

<211> 60

<212> PROTEIN

<213> Medicago truncatula

<400> 182

Met Ala Gln Ile Leu Met Phe Val Tyr Phe Leu Ile Ile Phe Leu Ser

1 5 10 15

Leu Phe Leu Val Glu Ser Ile Lys Ile Phe Thr Glu His Arg Cys Arg

20 25 30

Thr Asp Ala Asp Cys Pro Ala Arg Glu Leu Pro Glu Tyr Leu Lys Cys

35 40 45

Gln Gly Gly Met Cys Arg Leu Leu Ile Lys Lys Asp

50 55 60

<210> 183

<211> 56

<212> PROTEIN

<213> Medicago truncatula

<400> 183

Met Ala Arg Val Ile Ser Leu Phe Tyr Ala Leu Ile Ile Phe Leu Phe

1 5 10 15

Leu Phe Leu Val Ala Thr Asn Gly Asp Leu Ser Pro Cys Leu Arg Ser

20 25 30

Gly Asp Cys Ser Lys Asp Glu Cys Pro Ser His Leu Val Pro Lys Cys

35 40 45

Ile Gly Leu Thr Cys Tyr Cys Ile

50 55

<210> 184

<211> 62

<212> PROTEIN

<213> Medicago truncatula

<400> 184

Met Gln Arg Arg Lys Asn Met Ala Gln Ile Leu Leu Phe Ala Tyr Val

1 5 10 15

Phe Ile Ile Ser Ile Ser Leu Phe Leu Val Val Thr Asn Gly Val Lys

20 25 30

Ile Pro Cys Val Lys Asp Thr Asp Cys Pro Thr Leu Pro Cys Pro Leu

35 40 45

Tyr Ser Lys Cys Val Asp Gly Phe Cys Lys Met Leu Ser Ile

50 55 60

<210> 185

<211> 66

<212> PROTEIN

<213> Medicago truncatula

<400> 185

Met Asn His Ile Ser Lys Phe Val Tyr Ala Leu Ile Ile Phe Leu Ser

1 5 10 15

Val Tyr Leu Val Val Leu Asp Gly Arg Pro Val Ser Cys Lys Asp His

20 25 30

Tyr Asp Cys Arg Arg Lys Val Lys Ile Val Gly Cys Ile Phe Pro Gln

35 40 45

Glu Lys Pro Met Cys Ile Asn Ser Met Cys Thr Cys Ile Arg Glu Ile

50 55 60

ValPro

65

<210> 186

<211> 86

<212> PROTEIN

<213> Medicago truncatula

<400> 186

Met Lys Ser Gln Asn His Ala Lys Phe Ile Ser Phe Tyr Lys Asn Asp

1 5 10 15

Leu Phe Lys Ile Phe Gln Asn Asn Asp Ser His Phe Lys Val Phe Phe

20 25 30

Ala Leu Ile Ile Phe Leu Tyr Thr Tyr Leu His Val Thr Asn Gly Val

35 40 45

Phe Val Ser Cys Asn Ser His Ile His Cys Arg Val Asn Asn His Lys

50 55 60

Ile Gly Cys Asn Ile Pro Glu Gln Tyr Leu Leu Cys Val Asn Leu Phe

65 70 75 80

Cys Leu Trp Leu Asp Tyr

85

<210> 187

<211> 62

<212> PROTEIN

<213> Medicago truncatula

<400> 187

Met Thr Tyr Ile Ser Lys Val Val Tyr Ala Leu Ile Ile Phe Leu Ser

1 5 10 15

Ile Tyr Val Gly Val Asn Asp Cys Met Leu Val Thr Cys Glu Asp His

20 25 30

Phe Asp Cys Arg Gln Asn Val Gln Gln Val Gly Cys Ser Phe Arg Glu

35 40 45

Ile Pro Gln Cys Ile Asn Ser Ile Cys Lys Cys Met Lys Gly

50 55 60

<210> 188

<211> 63

<212> PROTEIN

<213> Medicago truncatula

<400> 188

Met Thr His Ile Ser Lys Phe Val Phe Ala Leu Ile Ile Phe Leu Ser

1 5 10 15

Ile Tyr Val Gly Val Asn Asp Cys Lys Arg Ile Pro Cys Lys Asp Asn

20 25 30

Asn Asp Cys Asn Asn Asn Trp Gln Leu Leu Ala Cys Arg Phe Glu Arg

35 40 45

Glu Val Pro Arg Cys Ile Asn Ser Ile Cys Lys Cys Met Pro Met

50 55 60

<210> 189

<211> 60

<212> PROTEIN

<213> Medicago truncatula

<400> 189

Met Val Gln Thr Pro Lys Leu Val Tyr Val Ile Val Leu Leu Leu Ser

1 5 10 15

Ile Phe Leu Gly Met Thr Ile Cys Asn Ser Ser Phe Ser His Phe Phe

20 25 30

Glu Gly Ala Cys Lys Ser Asp Lys Asp Cys Pro Lys Leu His Arg Ser

35 40 45

Asn Val Arg Cys Arg Lys Gly Gln Cys Val Gln Ile

50 55 60

<210> 190

<211> 77

<212> PROTEIN

<213> Medicago truncatula

<400> 190

Met Thr Lys Ile Leu Met Leu Phe Tyr Ala Met Ile Val Phe His Ser

1 5 10 15

Ile Phe Leu Val Ala Ser Tyr Thr Asp Glu Cys Ser Thr Asp Ala Asp

20 25 30

Cys Glu Tyr Ile Leu Cys Leu Phe Pro Ile Ile Lys Arg Cys Ile His

35 40 45

Asn His Cys Lys Cys Val Pro Met Gly Ser Ile Glu Pro Met Ser Thr

50 55 60

Ile Pro Asn Gly Val His Lys Phe His Ile Ile Asn Asn

65 70 75

<210> 191

<211> 64

<212> PROTEIN

<213> Medicago truncatula

<400> 191

Met Ala Lys Thr Leu Asn Phe Val Cys Ala Met Ile Leu Phe Ile Ser

1 5 10 15

Leu Phe Leu Val Ser Lys Asn Val Ala Leu Tyr Ile Ile Glu Cys Lys

20 25 30

Thr Asp Ala Asp Cys Pro Ile Ser Lys Leu Asn Met Tyr Asn Trp Arg

35 40 45

Cys Ile Lys Ser Ser Cys His Leu Tyr Lys Val Ile Gln Phe Met Val

50 55 60

<210> 192

<211> 72

<212> PROTEIN

<213> Medicago truncatula

<400> 192

Met Gln Lys Glu Lys Asn Met Ala Lys Thr Phe Glu Phe Val Tyr Ala

1 5 10 15

Met Ile Ile Phe Ile Leu Leu Phe Leu Val Glu Asn Asn Phe Ala Ala

20 25 30

Tyr Ile Ile Glu Cys Gln Thr Asp Asp Asp Cys Pro Lys Ser Gln Leu

35 40 45

Glu Met Phe Ala Trp Lys Cys Val Lys Asn Gly Cys His Leu Phe Gly

50 55 60

Met Tyr Glu Asp Asp Asp Asp Pro

65 70

<210> 193

<211> 57

<212> PROTEIN

<213> Medicago truncatula

<400> 193

Met Ala Ala Thr Arg Lys Phe Ile Tyr Val Leu Ser His Phe Leu Phe

1 5 10 15

Leu Phe Leu Val Thr Lys Ile Thr Asp Ala Arg Val Cys Lys Ser Asp

20 25 30

Lys Asp Cys Lys Asp Ile Ile Ile Tyr Arg Tyr Ile Leu Lys Cys Arg

35 40 45

Asn Gly Glu Cys Val Lys Ile Lys Ile

50 55

<210> 194

<211> 75

<212> PROTEIN

<213> Medicago truncatula

<400> 194

Met Gln Arg Leu Asp Asn Met Ala Lys Asn Val Lys Phe Ile Tyr Val

1 5 10 15

Ile Ile Leu Leu Leu Phe Ile Phe Leu Val Ile Ile Val Cys Asp Ser

20 25 30

Ala Phe Val Pro Asn Ser Gly Pro Cys Thr Thr Asp Lys Asp Cys Lys

35 40 45

Gln Val Lys Gly Tyr Ile Ala Arg Cys Arg Lys Gly Tyr Cys Met Gln

50 55 60

Ser Val Lys Arg Thr Trp Ser Ser Tyr Ser Arg

65 70 75

<210> 195

<211> 102

<212> PROTEIN

<213> Medicago truncatula

<400> 195

Met Lys Phe Ile Tyr Ile Met Ile Leu Phe Leu Ser Leu Phe Leu Val

1 5 10 15

Gln Phe Leu Thr Cys Lys Gly Leu Thr Val Pro Cys Glu Asn Pro Thr

20 25 30

Thr Cys Pro Glu Asp Phe Cys Thr Pro Pro Met Ile Thr Arg Cys Ile

35 40 45

Asn Phe Ile Cys Leu Cys Asp Gly Pro Glu Tyr Ala Glu Pro Glu Tyr

50 55 60

Asp Gly Pro Glu Pro Glu Tyr Asp His Lys Gly Asp Phe Leu Ser Val

65 70 75 80

Lys Pro Lys Ile Ile Asn Glu Asn Met Met Met Arg Glu Arg His Met

85 90 95

Met Lys Glu Ile Glu Val

100

<210> 196

<211> 59

<212> PROTEIN

<213> Medicago truncatula

<400> 196

Met Ala Gln Phe Leu Met Phe Ile Tyr Val Leu Ile Ile Phe Leu Tyr

1 5 10 15

Leu Phe Tyr Val Glu Ala Ala Met Phe Glu Leu Thr Lys Ser Thr Ile

20 25 30

Arg Cys Val Thr Asp Ala Asp Cys Pro Asn Val Val Lys Pro Leu Lys

35 40 45

Pro Lys Cys Val Asp Gly Phe Cys Glu Tyr Thr

50 55

<210> 197

<211> 70

<212> PROTEIN

<213> Medicago truncatula

<400> 197

Met Lys Met Arg Ile His Met Ala Gln Ile Ile Met Phe Phe Tyr Ala

1 5 10 15

Leu Ile Ile Phe Leu Ser Pro Phe Leu Val Asp Arg Arg Ser Phe Pro

20 25 30

Ser Ser Phe Val Ser Pro Lys Ser Tyr Thr Ser Glu Ile Pro Cys Lys

35 40 45

Ala Thr Arg Asp Cys Pro Tyr Glu Leu Tyr Tyr Glu Thr Lys Cys Val

50 55 60

Asp Ser Leu Cys Thr Tyr

65 70

<210> 198

<211> 41

<212> PROTEIN

<213> Medicago truncatula

<400> 198

Thr Arg Met Leu Thr Ile Pro Cys Thr Ser Asp Asp Asn Cys Pro Lys

1 5 10 15

Val Ile Ser Pro Cys His Thr Lys Cys Phe Asp Gly Phe Cys Gly Trp

20 25 30

Tyr Ile Glu Gly Ser Tyr Glu Gly Pro

35 40

<210> 199

<211> 69

<212> PROTEIN

<213> Medicago truncatula

<400> 199

Met Ala Gln Phe Leu Leu Phe Val Tyr Ser Leu Ile Ile Phe Leu Ser

1 5 10 15

Leu Phe Phe Gly Glu Ala Ala Phe Glu Arg Thr Glu Thr Arg Met Leu

20 25 30

Thr Ile Pro Cys Thr Ser Asp Asp Asn Cys Pro Lys Val Ile Ser Pro

35 40 45

Cys His Thr Lys Cys Phe Asp Gly Phe Cys Gly Trp Tyr Ile Glu Gly

50 55 60

Ser Tyr Glu Gly Pro

65

<210> 200

<211> 78

<212> PROTEIN

<213> Buchnera aphidicola

<400> 200

Met Lys Leu Leu His Gly Phe Leu Ile Ile Met Leu Thr Met His Leu

1 5 10 15

Ser Ile Gln Tyr Ala Tyr Gly Gly Pro Phe Leu Thr Lys Tyr Leu Cys

20 25 30

Asp Arg Val Cys His Lys Leu Cys Gly Asp Glu Phe Val Cys Ser Cys

35 40 45

Ile Gln Tyr Lys Ser Leu Lys Gly Leu Trp Phe Pro His Cys Pro Thr

50 55 60

Gly Lys Ala Ser Val Val Leu His Asn Phe Leu Thr Ser Pro

65 70 75

<210> 201

<211> 77

<212> PROTEIN

<213> Buchnera aphidicola

<400> 201

Met Lys Leu Leu Tyr Gly Phe Leu Ile Ile Met Leu Thr Ile His Leu

1 5 10 15

Ser Val Gln Tyr Phe Glu Ser Pro Phe Glu Thr Lys Tyr Asn Cys Asp

20 25 30

Thr His Cys Asn Lys Leu Cys Gly Lys Ile Asp His Cys Ser Cys Ile

35 40 45

Gln Tyr His Ser Met Glu Gly Leu Trp Phe Pro His Cys Arg Thr Gly

50 55 60

Ser Ala Ala Gln Met Leu His Asp Phe Leu Ser Asn Pro

65 70 75

<210> 202

<211> 86

<212> PROTEIN

<213> Buchnera aphidicola

<400> 202

Met Ser Val Arg Lys Asn Val Leu Pro Thr Met Phe Val Val Leu Leu

1 5 10 15

Ile Met Ser Pro Val Thr Pro Thr Ser Val Phe Ile Ser Ala Val Cys

20 25 30

Tyr Ser Gly Cys Gly Ser Leu Ala Leu Val Cys Phe Val Ser Asn Gly

35 40 45

Ile Thr Asn Gly Leu Asp Tyr Phe Lys Ser Ser Ala Pro Leu Ser Thr

50 55 60

Ser Glu Thr Ser Cys Gly Glu Ala Phe Asp Thr Cys Thr Asp His Cys

65 70 75 80

Leu Ala Asn Phe Lys Phe

85

<210> 203

<211> 69

<212> PROTEIN

<213> Buchnera aphidicola

<400> 203

Met Arg Leu Leu Tyr Gly Phe Leu Ile Ile Met Leu Thr Ile Tyr Leu

1 5 10 15

Ser Val Gln Asp Phe Asp Pro Thr Glu Phe Lys Gly Pro Phe Pro Thr

20 25 30

Ile Glu Ile Cys Ser Lys Tyr Cys Ala Val Val Cys Asn Tyr Thr Ser

35 40 45

Arg Pro Cys Tyr Cys Val Glu Ala Ala Lys Glu Arg Asp Gln Trp Phe

50 55 60

Pro Tyr Cys Tyr Asp

65

<210> 204

<211> 77

<212> PROTEIN

<213> Buchnera aphidicola

<400> 204

Met Arg Leu Leu Tyr Gly Phe Leu Ile Ile Met Leu Thr Ile His Leu

1 5 10 15

Ser Val Gln Asp Ile Asp Pro Asn Thr Leu Arg Gly Pro Tyr Pro Thr

20 25 30

Lys Glu Ile Cys Ser Lys Tyr Cys Glu Tyr Asn Val Val Cys Gly Ala

35 40 45

Ser Leu Pro Cys Ile Cys Val Gln Asp Ala Arg Gln Leu Asp His Trp

50 55 60

Phe Ala Cys Cys Tyr Asp Gly Gly Pro Glu Met Leu Met

65 70 75

<210> 205

<211> 108

<212> PROTEIN

<213> Buchnera aphidicola

<400> 205

Met Lys Leu Phe Val Val Val Val Leu Val Ala Val Gly Ile Met Phe

1 5 10 15

Val Phe Ala Ser Asp Thr Ala Ala Ala Pro Thr Asp Tyr Glu Asp Thr

20 25 30

Asn Asp Met Ile Ser Leu Ser Ser Leu Val Gly Asp Asn Ser Pro Tyr

35 40 45

Val Arg Val Ser Ser Ala Asp Ser Gly Gly Ser Ser Lys Thr Ser Ser

50 55 60

Lys Asn Pro Ile Leu Gly Leu Leu Lys Ser Val Ile Lys Leu Leu Thr

65 70 75 80

Lys Ile Phe Gly Thr Tyr Ser Asp Ala Ala Pro Ala Met Pro Pro Ile

85 90 95

Pro Pro Ala Leu Arg Lys Asn Arg Gly Met Leu Ala

100 105

<210> 206

<211> 178

<212> PROTEIN

<213> Buchnera aphidicola

<400> 206

Met Val Ala Cys Lys Val Ile Leu Ala Val Ala Val Val Phe Val Ala

1 5 10 15

Ala Val Gln Gly Arg Pro Gly Gly Glu Pro Glu Trp Ala Ala Pro Ile

20 25 30

Phe Ala Glu Leu Lys Ser Val Ser Asp Asn Ile Thr Asn Leu Val Gly

35 40 45

Leu Asp Asn Ala Gly Glu Tyr Ala Thr Ala Ala Lys Asn Asn Leu Asn

50 55 60

Ala Phe Ala Glu Ser Leu Lys Thr Glu Ala Ala Val Phe Ser Lys Ser

65 70 75 80

Phe Glu Gly Lys Ala Ser Ala Ser Asp Val Phe Lys Glu Ser Thr Lys

85 90 95

Asn Phe Gln Ala Val Val Asp Thr Tyr Ile Lys Asn Leu Pro Lys Asp

100 105 110

Leu Thr Leu Lys Asp Phe Thr Glu Lys Ser Glu Gln Ala Leu Lys Tyr

115 120 125

Met Val Glu His Gly Thr Glu Ile Thr Lys Lys Ala Gln Gly Asn Thr

130 135 140

Glu Thr Glu Lys Glu Ile Lys Glu Phe Phe Lys Lys Gln Ile Glu Asn

145 150 155 160

Leu Ile Gly Gln Gly Lys Ala Leu Gln Ala Lys Ile Ala Glu Ala Lys

165 170 175

Lys Ala

<210> 207

<211> 311

<212> PROTEIN

<213> Buchnera aphidicola

<400> 207

Met Lys Thr Ser Ser Ser Lys Val Phe Ala Ser Cys Val Ala Ile Val

1 5 10 15

Cys Leu Ala Ser Val Ala Asn Ala Leu Pro Val Gln Lys Ser Val Ala

20 25 30

Ala Thr Thr Glu Asn Pro Ile Val Glu Lys His Gly Cys Arg Ala His

35 40 45

Lys Asn Leu Val Arg Gln Asn Val Val Asp Leu Lys Thr Tyr Asp Ser

50 55 60

Met Leu Ile Thr Asn Glu Val Val Gln Lys Gln Ser Asn Glu Val Gln

65 70 75 80

Ser Ser Glu Gln Ser Asn Glu Gly Gln Asn Ser Glu Gln Ser Asn Glu

85 90 95

Gly Gln Asn Ser Glu Gln Ser Asn Glu Val Gln Ser Ser Glu His Ser

100 105 110

Asn Glu Gly Gln Asn Ser Lys Gln Ser Asn Glu Gly Gln Asn Ser Glu

115 120 125

Gln Ser Asn Glu Val Gln Ser Ser Glu His Ser Asn Glu Gly Gln Asn

130 135 140

Ser Glu Gln Ser Asn Glu Val Gln Ser Ser Glu His Ser Asn Glu Gly

145 150 155 160

Gln Asn Ser Lys Gln Ser Asn Glu Gly Gln Asn Ser Lys Gln Ser Asn

165 170 175

Glu Val Gln Ser Ser Glu His Trp Asn Glu Gly Gln Asn Ser Lys Gln

180 185 190

Ser Asn Glu Asp Gln Asn Ser Glu Gln Ser Asn Glu Gly Gln Asn Ser

195 200 205

Lys Gln Ser Asn Glu Gly Gln Asn Ser Lys Gln Ser Asn Glu Asp Gln

210 215 220

Asn Ser Glu Gln Ser Asn Glu Gly Gln Asn Ser Lys Gln Ser Asn Glu

225 230 235 240

Val Gln Ser Ser Glu Gln Ser Asn Glu Gly Gln Asn Ser Lys Gln Ser

245 250 255

Asn Glu Gly Gln Ser Ser Glu Gln Ser Asn Glu Gly Gln Asn Ser Lys

260 265 270

Gln Ser Asn Glu Val Gln Ser Pro Glu Glu His Tyr Asp Leu Pro Asp

275 280 285

Pro Glu Ser Ser Tyr Glu Ser Glu Ser Glu Thr Lys Gly Ser His Glu Ser

290 295 300

Gly Asp Asp Ser Glu His Arg

305 310

<210> 208

<211> 431

<212> PROTEIN

<213> Buchnera aphidicola

<400> 208

Met Lys Thr Ile Ile Leu Gly Leu Cys Leu Phe Gly Ala Leu Phe Trp

1 5 10 15

Ser Thr Gln Ser Met Pro Val Gly Glu Val Ala Pro Ala Val Pro Ala

20 25 30

Val Pro Ser Glu Ala Val Pro Gln Lys Gln Val Glu Ala Lys Pro Glu

35 40 45

Thr Asn Ala Ala Ser Pro Val Ser Asp Ala Lys Pro Glu Ser Asp Ser

50 55 60

Lys Pro Val Asp Ala Glu Val Lys Pro Thr Val Ser Glu Val Lys Ala

65 70 75 80

Glu Ser Glu Gln Lys Pro Ser Gly Glu Pro Lys Pro Glu Ser Asp Ala

85 90 95

Lys Pro Val Val Ala Ser Glu Ser Lys Pro Glu Ser Asp Pro Lys Pro

100 105 110

Ala Ala Val Val Glu Ser Lys Pro Glu Asn Asp Ala Val Ala Pro Glu

115 120 125

Thr Asn Asn Asp Ala Lys Pro Glu Asn Ala Ala Ala Pro Val Ser Glu

130 135 140

Asn Lys Pro Ala Thr Asp Ala Lys Ala Glu Thr Glu Leu Ile Ala Gln

145 150 155 160

Ala Lys Pro Glu Ser Lys Pro Ala Ser Asp Leu Lys Ala Glu Pro Glu

165 170 175

Ala Ala Lys Pro Asn Ser Glu Val Pro Val Ala Leu Pro Leu Asn Pro

180 185 190

Thr Glu Thr Lys Ala Thr Gln Gln Ser Val Glu Thr Asn Gln Val Glu

195 200 205

Gln Ala Ala Pro Ala Ala Ala Gln Ala Asp Pro Ala Ala Ala Pro Ala

210 215 220

Ala Asp Pro Ala Pro Ala Pro Ala Ala Ala Pro Val Ala Ala Glu Glu

225 230 235 240

Ala Lys Leu Ser Glu Ser Ala Pro Ser Thr Glu Asn Lys Ala Ala Glu

245 250 255

Glu Pro Ser Lys Pro Ala Glu Gln Gln Ser Ala Lys Pro Val Glu Asp

260 265 270

Ala Val Pro Ala Ala Ser Glu Ile Ser Glu Thr Lys Val Ser Pro Ala

275 280 285

Val Pro Ala Val Pro Glu Val Pro Ala Ser Pro Ser Ala Pro Ala Val

290 295 300

Ala Asp Pro Val Ser Ala Pro Glu Ala Glu Lys Asn Ala Glu Pro Ala

305 310 315 320

Lys Ala Ala Asn Ser Ala Glu Pro Ala Val Gln Ser Glu Ala Lys Pro

325 330 335

Ala Glu Asp Ile Gln Lys Ser Gly Ala Val Val Ser Ala Glu Asn Pro

340 345 350

Lys Pro Val Glu Glu Gln Lys Pro Ala Glu Val Ala Lys Pro Ala Glu

355 360 365

Gln Ser Lys Ser Glu Ala Pro Ala Glu Ala Pro Lys Pro Thr Glu Gln

370 375 380

Ser Ala Ala Glu Glu Pro Lys Lys Pro Glu Ser Ala Asn Asp Glu Lys

385 390 395 400

Lys Glu Gln His Ser Val Asn Lys Arg Asp Ala Thr Lys Glu Lys Lys

405 410 415

Pro Thr Asp Ser Ile Met Lys Lys Gln Lys Gln Lys Lys Ala Asn

420 425 430

<210> 209

<211> 160

<212> PROTEIN

<213> Buchnera aphidicola

<400> 209

Met Asn Gly Lys Ile Val Leu Cys Phe Ala Val Val Phe Ile Gly Gln

1 5 10 15

Ala Met Ser Ala Ala Thr Gly Thr Thr Pro Glu Val Glu Asp Ile Lys

20 25 30

Lys Val Ala Glu Gln Met Ser Gln Thr Phe Met Ser Val Ala Asn His

35 40 45

Leu Val Gly Ile Thr Pro Asn Ser Ala Asp Ala Gln Lys Ser Ile Glu

50 55 60

Lys Ile Arg Thr Ile Met Asn Lys Gly Phe Thr Asp Met Glu Thr Glu

65 70 75 80

Ala Asn Lys Met Lys Asp Ile Val Arg Lys Asn Ala Asp Pro Lys Leu

85 90 95

Val Glu Lys Tyr Asp Glu Leu Glu Lys Glu Leu Lys Lys His Leu Ser

100 105 110

Thr Ala Lys Asp Met Phe Glu Asp Lys Val Val Lys Pro Ile Gly Glu

115 120 125

Lys Val Glu Leu Lys Lys Ile Thr Glu Asn Val Ile Lys Thr Thr Lys

130 135 140

Asp Met Glu Ala Thr Met Asn Lys Ala Ile Asp Gly Phe Lys Lys Gln

145 150 155 160

<210> 210

<211> 415

<212> PROTEIN

<213> Buchnera aphidicola

<400> 210

Met His Leu Phe Leu Ala Leu Gly Leu Phe Ile Val Cys Gly Met Val

1 5 10 15

Asp Ala Thr Phe Tyr Asn Pro Arg Ser Gln Thr Phe Asn Gln Leu Met

20 25 30

Glu Arg Arg Gln Arg Ser Ile Pro Ile Pro Tyr Ser Tyr Gly Tyr His

35 40 45

Tyr Asn Pro Ile Glu Pro Ser Ile Asn Val Leu Asp Ser Leu Ser Glu

50 55 60

Gly Leu Asp Ser Arg Ile Asn Thr Phe Lys Pro Ile Tyr Gln Asn Val

65 70 75 80

Lys Met Ser Thr Gln Asp Val Asn Ser Val Pro Arg Thr Gln Tyr Gln

85 90 95

Pro Lys Asn Ser Leu Tyr Asp Ser Glu Tyr Ile Ser Ala Lys Asp Ile

100 105 110

Pro Ser Leu Phe Pro Glu Glu Asp Ser Tyr Asp Tyr Lys Tyr Leu Gly

115 120 125

Ser Pro Leu Asn Lys Tyr Leu Thr Arg Pro Ser Thr Gln Glu Ser Gly

130 135 140

Ile Ala Ile Asn Leu Val Ala Ile Lys Glu Thr Ser Val Phe Asp Tyr

145 150 155 160

Gly Phe Pro Thr Tyr Lys Ser Pro Tyr Ser Ser Asp Ser Val Trp Asn

165 170 175

Phe Gly Ser Lys Ile Pro Asn Thr Val Phe Glu Asp Pro Gln Ser Val

180 185 190

Glu Ser Asp Pro Asn Thr Phe Lys Val Ser Ser Pro Thr Ile Lys Ile

195 200 205

Val Lys Leu Leu Pro Glu Thr Pro Glu Gln Glu Ser Ile Ile Thr Thr

210 215 220

Thr Lys Asn Tyr Glu Leu Asn Tyr Lys Thr Thr Gln Glu Thr Pro Thr

225 230 235 240

Glu Ala Glu Leu Tyr Pro Ile Thr Ser Glu Glu Phe Gln Thr Glu Asp

245 250 255

Glu Trp His Pro Met Val Pro Lys Glu Asn Thr Thr Lys Asp Glu Ser

260 265 270

Ser Phe Ile Thr Thr Glu Glu Pro Leu Thr Glu Asp Lys Ser Asn Ser

275 280 285

Ile Thr Ile Glu Lys Thr Gln Thr Glu Asp Glu Ser Asn Ser Ile Glu

290 295 300

Phe Asn Ser Ile Arg Thr Glu Glu Lys Ser Asn Ser Ile Thr Glu

305 310 315 320

Glu Asn Gln Lys Glu Asp Asp Glu Ser Met Ser Thr Thr Ser Gln Glu

325 330 335

Thr Thr Thr Ala Phe Asn Leu Asn Asp Thr Phe Asp Thr Asn Arg Tyr

340 345 350

Ser Ser Ser His Glu Ser Leu Met Leu Arg Ile Arg Glu Leu Met Lys

355 360 365

Asn Ile Ala Asp Gln Gln Asn Lys Ser Gln Phe Arg Thr Val Asp Asn

370 375 380

Ile Pro Ala Lys Ser Gln Ser Asn Leu Ser Ser Asp Glu Ser Thr Asn

385 390 395 400

Gln Gln Phe Glu Pro Gln Leu Val Asn Gly Ala Asp Thr Tyr Lys

405 410 415

<210> 211

<211> 126

<212> PROTEIN

<213> Sitophilus zeamais

<400> 211

Met Thr Arg Thr Met Leu Phe Leu Ala Cys Val Ala Ala Leu Tyr Val

1 5 10 15

Cys Ile Ser Ala Thr Ala Gly Lys Pro Glu Glu Phe Ala Lys Leu Ser

20 25 30

Asp Glu Ala Pro Ser Asn Asp Gln Ala Met Tyr Glu Ser Ile Gln Arg

35 40 45

Tyr Arg Arg Phe Val Asp Gly Asn Arg Tyr Asn Gly Gly Gln Gln Gln

50 55 60

Gln Gln Gln Pro Lys Gln Trp Glu Val Arg Pro Asp Leu Ser Arg Asp

65 70 75 80

Gln Arg Gly Asn Thr Lys Ala Gln Val Glu Ile Asn Lys Lys Gly Asp

85 90 95

Asn His Asp Ile Asn Ala Gly Trp Gly Lys Asn Ile Asn Gly Pro Asp

100 105 110

Ser His Lys Asp Thr Trp His Val Gly Gly Ser Val Arg Trp

115 120 125

<210> 212

<211> 220

<212> PROTEIN

<213> Acyrthosiphon pisum

<400> 212

Met Lys Glu Thr Thr Val Val Trp Ala Lys Leu Phe Leu Ile Leu Ile

1 5 10 15

Ile Leu Ala Lys Pro Leu Gly Leu Lys Ala Val Asn Glu Cys Lys Arg

20 25 30

Leu Gly Asn Asn Ser Cys Arg Ser His Gly Glu Cys Cys Ser Gly Phe

35 40 45

Cys Phe Ile Glu Pro Gly Trp Ala Leu Gly Val Cys Lys Arg Leu Gly

50 55 60

Thr Pro Lys Lys Ser Asp Asp Ser Asn Asn Gly Lys Asn Ile Glu Lys

65 70 75 80

Asn Asn Gly Val His Glu Arg Ile Asp Asp Val Phe Glu Arg Gly Val

85 90 95

Cys Ser Tyr Tyr Lys Gly Pro Ser Ile Thr Ala Asn Gly Asp Val Phe

100 105 110

Asp Glu Asn Glu Met Thr Ala Ala His Arg Thr Leu Pro Phe Asn Thr

115 120 125

Met Val Lys Val Glu Gly Met Gly Thr Ser Val Val Val Lys Ile Asn

130 135 140

Asp Arg Lys Thr Ala Ala Asp Gly Lys Val Met Leu Leu Ser Arg Ala

145 150 155 160

Ala Ala Glu Ser Leu Asn Ile Asp Glu Asn Thr Gly Pro Val Gln Cys

165 170 175

Gln Leu Lys Phe Val Leu Asp Gly Ser Gly Cys Thr Pro Asp Tyr Gly

180 185 190

Asp Thr Cys Val Leu His Glu Cys Cys Ser Gln Asn Cys Phe Arg

195 200 205

Glu Met Phe Ser Asp Lys Gly Phe Cys Leu Pro Lys

210 215 220

<210> 213

<211> 16

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Penetratin

<400> 213

Arg Gln Ile Lys Ile Trp Phe Gln Asn Arg Arg Met Lys Trp Lys Lys

1 5 10 15

<210> 214

<211> 13

<212> PROTEIN

<213> Human Immunodeficiency Virus 1

<400> 214

Gly Arg Lys Lys Arg Arg Gln Arg Arg Arg Pro Pro Gln

1 5 10

<210> 215

<211> 18

<212> PROTEIN

<213> Artificial sequence

<220>

<223> pVEC

<400> 215

Leu Leu Ile Ile Leu Arg Arg Arg Ile Arg Lys Gln Ala His Ala His

1 5 10 15

Ser Lys

<210> 216

<211> 27

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Transportan

<400> 216

Gly Trp Thr Leu Asn Ser Ala Gly Tyr Leu Leu Gly Lys Ile Asn Leu

1 5 10 15

Lys Ala Leu Ala Ala Leu Ala Lys Lys Ile Leu

20 25

<210> 217

<211> 27

<212> PROTEIN

<213> Artificial sequence

<220>

<223> MPG

<400> 217

Gly Ala Leu Phe Leu Gly Phe Leu Gly Ala Ala Gly Ser Thr Met Gly

1 5 10 15

Ala Trp Ser Gln Pro Lys Lys Lys Arg Lys Val

20 25

<210> 218

<211> 21

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Pep-1

<400> 218

Lys Glu Thr Trp Trp Glu Thr Trp Trp Thr Glu Trp Ser Gln Pro Lys

1 5 10 15

Lys Lys Arg Lys Val

twenty

<210> 219

<211> 18

<212> PROTEIN

<213> Artificial sequence

<220>

<223> MAP

<400> 219

Lys Leu Ala Leu Lys Leu Ala Leu Lys Ala Leu Lys Ala Ala Leu Lys

1 5 10 15

Leu Ala

<210> 220

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<223> R6W3

<400> 220

Arg Arg Trp Trp Arg Arg Trp Arg Arg

fifteen

<210> 221

<211> 13

<212> PROTEIN

<213> Pandinus imperator

<400> 221

Phe Leu Ser Thr Ile Trp Asn Gly Ile Lys Gly Leu Leu

1 5 10

<210> 222

<211> 13

<212> PROTEIN

<213>Urodacus yaschenkoi

<400> 222

Ile Leu Ser Ala Ile Trp Ser Gly Ile Lys Ser Leu Phe

1 5 10

<210> 223

<211> 13

<212> PROTEIN

<213> Scorpiops tibetanus

<400> 223

Leu Trp Gly Lys Leu Trp Glu Gly Val Lys Ser Leu Ile

1 5 10

<210> 224

<211> 22

<212> PROTEIN

<213> Apostichopus japonicus

<400> 224

Phe Pro Phe Leu Lys Leu Ser Leu Lys Ile Pro Lys Ser Ala Ile Lys

1 5 10 15

Ser Ala Ile Lys Arg Leu

twenty

<210> 225

<211> 13

<212> PROTEIN

<213>Urodacus yaschenkoi

<400> 225

Ile Leu Ser Ala Ile Trp Ser Gly Ile Lys Gly Leu Leu

1 5 10

<210> 226

<211> 27

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Uy192 + cell-penetrating peptide

<400> 226

Tyr Gly Arg Lys Lys Arg Arg Gln Arg Arg Arg Phe Leu Ser Thr Ile

1 5 10 15

Trp Asn Gly Ile Lys Gly Leu Leu Phe Ala Met

20 25

<210> 227

<211> 20

<212> DNA

<213> Artificial sequence

<220>

<223> Universal bacterial primer 27F

<400> 227

agagtttgat cmtggctcag 20

<210> 228

<211> 16

<212> DNA

<213> Artificial sequence

<220>

<223> 1492R Universal Bacterial Primer

<400> 228

taccttgtta cgactt 16

<---

Claims (6)

1. A method for improving the nutritional profile of an insect, comprising: delivering an effective amount of methionine-producing bacteria to the insect, the insect being at the larval stage. 2. The method of claim 1 wherein the insect is a cricket, grasshopper or locust. 3. The method of claim 1 or 2, wherein the delivery comprises delivering the composition to at least one habitat in which the insect grows, lives, breeds, or feeds. 4. The method according to any one of paragraphs. 1-3 wherein the methionine producing bacteria are delivered into insect edible compositions for ingestion by insects. 5. The method according to any one of paragraphs. 1-4 wherein the methionine producing bacteria are formulated with an agriculturally acceptable carrier in the form of a liquid, solid, aerosol, paste, gel or gaseous composition. 6. The method of claim 5 wherein the carrier is a seed coater.

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