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WO2006018205A2 - Nouveaux genes essentiels de bacillus licheniformis et procede de production biotechnologiques ameliores, bases sur ceux-ci - Google Patents

Nouveaux genes essentiels de bacillus licheniformis et procede de production biotechnologiques ameliores, bases sur ceux-ci Download PDF

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Publication number
WO2006018205A2
WO2006018205A2 PCT/EP2005/008683 EP2005008683W WO2006018205A2 WO 2006018205 A2 WO2006018205 A2 WO 2006018205A2 EP 2005008683 W EP2005008683 W EP 2005008683W WO 2006018205 A2 WO2006018205 A2 WO 2006018205A2
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identity
seq
nucleotide sequence
amino acid
acid sequence
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WO2006018205A3 (fr
Inventor
Jörg FEESCHE
Stefan Evers
Cornelius Bessler
Martina Plath
Armin Ehrenreich
Birgit Veith
Heiko Liesegang
Anke Henne
Christina Herzberg
Gerhard Gottschalk
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Henkel AG and Co KGaA
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Henkel AG and Co KGaA
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
    • C07K14/32—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Bacillus (G)

Definitions

  • the present invention relates to 150 new essential genes and their gene products of Bacillus licheniformis and sufficiently similar genes and proteins as well as auf ⁇ building, insofar improved biotechnological production process by Mikro ⁇ organisms, as each of these genes is suitable as a selection marker.
  • the present invention is in the field of biotechnology, in particular the production of recyclables by fermentation of microorganisms which are capable of forming the valuable substances of interest.
  • microorganisms which are capable of forming the valuable substances of interest.
  • These include, for example, the production of low molecular weight compounds, for example of food supplements or pharmaceutically relevant compounds, or of proteins, for which, due to their diversity, there is again a large field of technical application.
  • the metabolic properties of the microorganisms in question are utilized and / or modified for the production of valuable substances;
  • cells are used which express the genes of the proteins of interest (so-called transgenes).
  • GMOs genetically modified organisms
  • Expression systems are developed and developed essentially on the basis of two fundamentally different genetic constructions.
  • the gene for the protein to be produced becomes the chromosome of the host organism integrated.
  • Such constructs are very stable without selection for the presence of an additional marker gene (see below).
  • a major disadvantage is that only one copy of the gene is present in the host and the integration of further copies to increase the product formation rate on the gene dose effect designed methodically very expensive. This prior art is briefly illustrated below.
  • European Patent EP 284126 B1 solves the problem of stable multiple integration by introducing into the cell multiple gene copies containing intervening endogenous and essential chromosomal DNA segments.
  • the patent application DD 277467 A1 discloses a process for the production of extracellular enzymes which is based on the stable, advantageously multiple integration of the genes coding for the enzyme of interest into the bacterial chromosome. Integration takes place via homologous areas. To control successful integration events, an erythromycin gene contained on the plasmid serves to inactivate upon successful integration.
  • the integration into the chromosome can take place via a single or double crossing-over via the gene for thymidylate synthetase.
  • the latter allows the control of this procedure, because in a simple crossing-over, the thy activity is preserved, while it is lost in a double crossing-over, that is, this auxotrophy is achieved.
  • a simple crossing-over in this particular system is associated with a resistance to the antibiotic trimethoprim, with a double the corresponding sensitivity.
  • the application WO 96/23073 A1 discloses a transposon-based system for integrating multiple copies of a gene of interest into the bacterial chromosome, which is characterized in that the marker gene of the plasmid is deleted by the integration and the resulting strains are thus free from a resistance are markers. Again, this marker requires a marker only to control the construction of the bacterial strain of interest.
  • the second approach to constructing producer strains is to transfer the gene of interest to an autonomously replicating element, for example a plasmid, and thereby transfer it into the host organism.
  • an autonomously replicating element for example a plasmid
  • the usually high number of plasmid copies per cell has an effect on the gene dosage effect.
  • a disadvantage is the fact that over the entire culture time a Selek ⁇ tion pressure must be exercised to keep the plasmids in the cells. Standard ⁇ this is done via the addition of antibiotics in the culture medium, while genes that confer resistance to the substances in question, are presented on the plasmids. Thus, only the cells are able to grow, which have the plasmids in sufficient numbers.
  • antibiotic resistance as a selection marker has increasingly come up against criticism in recent years.
  • the use of antibiotics is quite expensive, especially if the resistance is based on an enzyme degrading the antibiotic and therefore the substance in question must be supplied during the entire cultivation.
  • their widespread use in particular in other fields of technology, contributes to the spread of the resistance genes to other strains, even to pathological strains. This already leads to considerable difficulties, for example, in medical hygiene and in particular in the treatment of infectious diseases by so-called multi-resistant, human-pathogenic strains.
  • auxotrophy that is a targeted metabolic defect that makes the cells in question dependent on the supply of certain metabolic products.
  • Auxotrophic strains will then be paired with the transgene of interest to cure this auxotrophy. If lost, they would simultaneously lose their viability under appropriate culture conditions, resulting in the desired selection of the auxotrophic producer strains.
  • auxotrophies so far very problematic, since in industrial fermentation almost all necessary substrates are available in sufficient quantities and the cells in question can compensate for the lack of synthesis of a particular compound on the inclusion of this same compound from the nutrient medium ,
  • the application EP 251579 A2 offers the solution of using as host strains those which are deficient in terms of the thymidylate synthase gene essential for nucleotide metabolism. Accordingly, the gene can be made available for this function (thyA from Escherichia coli K12) via a vector and can cure the gene defect. If this vector additionally carries the gene for the protein of interest, an antibiotic-like selection of the producer cells occurs.
  • auxotrophic markers have so far only led to very sparse results in terms of the complex nutrient media generally used in industry. Because these usually contain numerous low molecular weight compounds such as nucleotides, vitamins or amino acids, over which such auxotrophies, that is, relatively simple metabolic defects are compensated. It would therefore be desirable to be able to select, which works in principle like an auxotrophy, but does not rely on such metabolic defects that are compensated by the commonly used complex nutrient media. It was therefore the task of developing further selection systems that are as easy to handle as the selection of an antibiotic, but do without expensive and possibly environmentally harmful substances. They should be applicable on an industrial scale. They should build on such genes, the absence of which can not be compensated for by impurities in industrial media.
  • this meant identifying essential genes and providing tools for the desired genetic modification by identifying the associated nucleotide sequences.
  • each of the 150 proteins found in the sequence listing for B.licheniformis including the respectively sufficiently homologous proteins; Further independent solutions to the problem are the associated nucleic acids, likewise including the respectively sufficiently homologous nucleic acids.
  • the respective nucleic acid and amino acid sequences are fully set forth in the Sequence Listing of the present application.
  • the said sub-aspect of the task for the development of such essential proteins for which there are technical applications due to their biochemical properties is also met.
  • the associated proteins can now be produced selectively via the nucleotide sequences provided according to the invention and are thus available to the relevant technical application options.
  • the subject nucleic acids may be used to identify related genes or promoters naturally associated therewith.
  • an initiator protein of the chromosomal replication DnaA having an amino acid sequence which corresponds to the amino acid sequence shown in SEQ ID NO. 2 has at least 95% identity and more preferably at least 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid dnaB coding for a membrane attachment protein of the chromosomal replication initiation having a nucleotide sequence which corresponds to that shown in SEQ ID NO. 3 nucleotide sequence has at least 74% identity and more preferably at least 75%, 80%, 85%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • a membrane attachment protein of the chromosomal replication initiation DnaB having an amino acid sequence identical to that shown in SEQ ID NO. 4 has at least 70% identity and more preferably at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid dnaC encoding a replicative DNA helicase, having a nucleotide sequence corresponding to that shown in SEQ ID NO. 5 nucleotide sequence has at least 87% identity and more preferably at least 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • a replicative DNA helicase DnaC having an amino acid sequence identical to that shown in SEQ ID NO. 6 amino acid sequence has at least 95% identity and increasingly preferably at least 96%, 97%, 98%, 99% and particularly preferably 100% identity.
  • nucleic acid dnaD encoding a DNA replication protein having a nucleotide sequence corresponding to that shown in SEQ ID NO. At least 77% identity and more preferably at least 80%, 85%, 87.5%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity having;
  • a DNA replication protein DnaD having an amino acid sequence corresponding to that shown in SEQ ID NO. 8 has at least 75% identity and more preferably at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid dnaE coding for an alpha subunit of a DNA polymerase III (DNA polymerase III alpha subunit, E.C. 2.7.7.7) having a nucleotide sequence which corresponds to that shown in SEQ ID NO. 9 nucleotide sequence has at least 74% identity, and more preferably at least 75%, 80%, 85%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • DNA polymerase III alpha subunit of a DNA polymerase III (DNA polymerase III alpha subunit; E.C. 2.7.7.7) DnaE having an amino acid sequence which corresponds to the amino acid sequence shown in SEQ ID NO. At least 74% identity, and more preferably at least 75%, 80%, 85%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • a nucleic acid dnaG encoding a DNA primase (DNA primase, E.C. 2.7.7.-) having a nucleotide sequence corresponding to that shown in SEQ ID NO. 11 nucleotide sequence has at least 76% identity, and more preferably at least 80%, 85%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • DNA primase (DNA primase, E.C. 2.7.7.) DnaG having an amino acid sequence identical to that shown in SEQ ID NO. 12 has at least 79% identity and more preferably at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid dnal coding for a primosome protein having a nucleotide sequence which corresponds to that shown in SEQ ID NO. 13 nucleotide sequence has at least 74% identity, and more preferably at least 75%, 80%, 85%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • a primosome protein Dnal having an amino acid sequence corresponding to that shown in SEQ ID NO. 14 has at least 74% identity, and more preferably at least 75%, 80%, 85%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity;
  • nucleic acid dnaN coding for a beta-chain of a DNA polymerase III (DNA polymerase IM beta chain; EC 2.7.7.7), having a nucleotide sequence which corresponds to that shown in SEQ ID NO. 15 nucleotide sequence has at least 82% identity, and more preferably at least 85%, 87.5%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity
  • a beta-chain of a DNA polymerase III (DNA polymerase III beta chain; EC 2.7.7.7) DnaN, having an amino acid sequence identical to that shown in SEQ ID NO. 16 has at least 89% identity and more preferably at least 90%, 92.2%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid dnaX coding for a chain III of a DNA-dependent DNA polymerase III (DNA-directed DNA polymerase III chain, E.C., 2.7.7.7) having a nucleotide sequence which corresponds to that shown in SEQ ID NO. At least 80% identity and more preferably at least 85%, 87.5%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity
  • a chain III of a DNA-dependent DNA polymerase III (DNA-directed DNA polymerase III chain, E.C., 2.7.7.7) DnaX having an amino acid sequence which corresponds to the amino acid sequence shown in SEQ ID NO. At least 85% identity, and more preferably at least 90%, 92.2%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid holB coding for a delta subunit of a DNA polymerase III (DNA polymerase III, delta 'subunit, E.C.2.7.7.7), having a nucleotide sequence which corresponds to that shown in SEQ ID NO. 19 nucleotide sequence has at least 84% identity, and more preferably at least 85%, 87.5%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • DNA polymerase III delta 1 subunit of a DNA polymerase III
  • HoIB having an amino acid sequence which corresponds to the amino acid sequence shown in SEQ ID NO. 20 has at least 85% identity, and more preferably at least 87.5%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity;
  • a nucleic acid HgA coding for a DNA ligase (DNA ligase, E.C. 6.5.1.2), having a nucleotide sequence which corresponds to that shown in SEQ ID NO. 21 has at least 81% identity and more preferably at least 85%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • LigA having an amino acid sequence which corresponds to the amino acid sequence shown in SEQ ID NO. 22 has at least 85% identity and more preferably at least 87.5%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • a nucleic acid pcrA coding for an ATP-dependent DNA helicase (ATP-dependent DNA helicase; EC 3.6.1.-) having a nucleotide sequence which corresponds to that shown in SEQ ID NO. 23 has at least 82% identity and more preferably at least 85%, 87.5%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • an ATP-dependent DNA helicase (ATP-dependent DNA helicase; E.C. 3.6.1.) PcrA, having an amino acid sequence corresponding to that shown in SEQ ID NO. 24 has at least 87% identity and more preferably at least 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid polC (pollll) coding for a PolC-type DNA polymerase III (DNA polymerase III, po / C-type), having a nucleotide sequence which corresponds to that shown in SEQ ID NO. 25 nucleotide sequence has at least 82% identity, and more preferably at least 85%, 87.5%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • a PolC-type DNA polymerase III (DNA polymerase III, po / C-type) PoIC having an amino acid sequence which corresponds to the amino acid sequence shown in SEQ ID NO. 26 has at least 89% identity and more preferably at least 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • a nucleic acid priA coding for a primosomal replication factor Y (primosomal protein N 1 ) having a nucleotide sequence which corresponds to that shown in SEQ ID NO. 27 has at least 76% identity and more preferably at least 80%, 85%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • a primosomal replication factor Y (primosomal protein N ') PriA having an amino acid sequence identical to that shown in SEQ ID NO. At least 79% identity and more preferably at least 80%, 82.5%, 85%, 87.5%, 90%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity having;
  • nucleic acid ssb coding for a single-stranded DNA-binding protein with a nucleotide sequence which corresponds to that shown in SEQ ID NO. 29 has at least 85% identity and more preferably at least 87.5%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • a nucleic acid gyrA coding for a subunit A of a DNA gyrase (DNA gyrase subunit A, E.C. 5.99.1.3), having a nucleotide sequence which corresponds to that shown in SEQ ID NO. 31 has at least 82% identity and more preferably at least 85%, 87.5%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • GyrA having an amino acid sequence identical to that shown in SEQ ID NO. 32 has at least 89% identity and more preferably at least 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid gyrB coding for a subunit B of a DNA gyrase (DNA gyrase subunit B, E.C. 5.99.1.3), having a nucleotide sequence which corresponds to that shown in SEQ ID NO. At least 82% identity, and more preferably at least 85%, 87.5%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • GyrB having an amino acid sequence identical to that shown in SEQ ID NO. Has at least 89% identity, and more preferably at least 90%, 92.5%, 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity;
  • a nucleic acid hbs coding for a DNA binding protein HU (DNA-binding protein HU), having a nucleotide sequence which corresponds to that shown in SEQ ID NO. 35 has at least 95% identity and more preferably at least 96%, 97%, 98%, 99% and most preferably 100% identity;
  • DNA binding protein HU (DNA-binding protein HU) Hbs having an amino acid sequence which corresponds to the amino acid sequence shown in SEQ ID NO. 36 has at least 97% identity and, more preferably, at least 97.5%, 98%, 98.5%, 99%, 99.5%, and most preferably 100% identity;
  • nucleic acid parC coding for a subunit A of a topoisomerase IV (topoisomerase IV 1 subunit A; EC 5.99.1.-), having a nucleotide sequence which corresponds to that shown in SEQ ID NO. 37 has at least 79% identity and more preferably at least 80%, 85%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity; a subunit A of a topoisomerase IV (topoisomerase IV, subunit A; EC 5.99.1.) ParC having an amino acid sequence which corresponds to the amino acid sequence shown in SEQ ID NO. At least 83% identity, and more preferably at least 85%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid parE coding for a subunit B of a topoisomerase IV (topoisomerase IV, subunit B, E.C. 5.99.1.-) having a nucleotide sequence which corresponds to that shown in SEQ ID NO. At least 84% identity, and more preferably at least 85%, 87.5%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • a subunit B of a topoisomerase IV (topoisomerase IV, subunit B, E.C. 5.99.1.-) ParE, having an amino acid sequence corresponding to that shown in SEQ ID NO. 40 has at least 94% identity and more preferably at least 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid smc coding for a factor for chromosome condensation and segregation having a nucleotide sequence which corresponds to that shown in SEQ ID NO. At least 76% identity and more preferably at least 80%, 85%, 87.5%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity having;
  • a chromosome condensation and segregation factor Smc having an amino acid sequence identical to that shown in SEQ ID NO. 42 has at least 81% identity and more preferably at least 85%, 87.5%, 90%, 92.5%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid topA encoding a DNA topoisomerase I (DNA topoisomerase I; E.C. 5.99.1.2) having a nucleotide sequence corresponding to that shown in SEQ ID NO. 43 nucleotide sequence has at least 83% identity and increasingly preferably at least 85%, 87.5%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • TopA having an amino acid sequence which corresponds to the amino acid sequence shown in SEQ ID NO. 44 has at least 91% identity and, more preferably, at least 92.5%, 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity;
  • an alpha subunit of a modification methylase (modification methylase, alpha subunit, E.C. 2.1.1.73) YdiO (Aqul) having an amino acid sequence corresponding to that shown in SEQ ID NO. At least 15% identity, and more preferably at least 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 92.5%, 95%, 96%, 97%, 98 %, 99%, and most preferably 100% identity;
  • nucleic acid rpoA coding for an alpha-chain of a DNA-dependent RNA polymerase (DNA 2.7.7.6), having a nucleotide sequence which corresponds to the nucleotide sequence given in SEQ ID NO % Identity, and more preferably at least 94%, 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity;
  • nucleic acid rpoB coding for a beta chain of a DNA-dependent RNA polymerase (E.C. 2.7.7.6), having a nucleotide sequence which corresponds to that shown in SEQ ID NO. 49 nucleotide sequence has at least 89% identity, and more preferably at least 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • RpoB having an amino acid sequence corresponding to that shown in SEQ ID NO. 50 has at least 94% identity and more preferably at least 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • a nucleic acid rpoC coding for a beta-chain of a DNA-dependent RNA polymerase (EC 2.7.7.6), having a nucleotide sequence which corresponds to that shown in SEQ ID NO. 51 has at least 88% identity and more preferably at least 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • a beta 'chain of a DNA-dependent RNA polymerase beta' chain (EC 2.7.7.6) RpoC having an amino acid sequence which corresponds to the amino acid sequence shown in SEQ ID NO. At least 93% identity, and more preferably at least 94%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity of the indicated amino acid sequence;
  • RNA polymerase major sigma-43 factor RNA polymerase major sigma-43 factor; sigma-A
  • RNA polymerase major sigma-43 factor RNA polymerase major sigma-43 factor; sigma-A
  • RNA polymerase major sigma-43 factor RNA polymerase major sigma-43 factor; sigma-A
  • SigA having an amino acid sequence corresponding to that shown in SEQ ID NO. 54 has at least 94% identity and, more preferably, at least 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity;
  • nucleic acid cca encoding a polyA polymerase (polyA polymerase) having a nucleotide sequence corresponding to that shown in SEQ ID NO. At least 66% identity, and more preferably at least 70%, 75%, 80%, 85%, 87.5%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99%, of the nucleotide sequence indicated. and most preferably 100% identity;
  • polyA polymerase (polyA polymerase) Cca, having an amino acid sequence corresponding to that shown in SEQ ID NO. At least 60% identity, and more preferably at least 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid cspR coding for a rRNA methylase having a nucleotide sequence which corresponds to that shown in SEQ ID NO. At least 79% identity, and more preferably at least 80%, 85%, 87.5%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity having;
  • rRNA methylase (rRNA methylase) CspR having an amino acid sequence which corresponds to the amino acid sequence shown in SEQ ID NO. At least 84% identity, and more preferably at least 85%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid rnc coding for a ribonuclease III having a nucleotide sequence which corresponds to that shown in SEQ ID NO. 59 indicated nucleotide sequence at least 86% identity, and more preferably at least 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • ribonuclease III (ribonuclease III) Rnc having an amino acid sequence which corresponds to the amino acid sequence shown in SEQ ID NO. At least 93% identity, and more preferably at least 94%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid rnpA coding for a ribonuclease P (ribonuclease P, E.C. 3.1.26.5), having a nucleotide sequence which corresponds to that shown in SEQ ID NO. At least 65% identity, and more preferably at least 70%, 75%, 80%, 85%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100%. Having identity;
  • RnpA having an amino acid sequence corresponding to that shown in SEQ ID NO. 62 has at least 69% identity and more preferably at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid trmD encoding a methyltransferase (methyltransferase; E.C. 2.1.1.31) having a nucleotide sequence corresponding to that shown in SEQ ID NO. At least 77% identity and more preferably at least 80%, 85%, 87.5%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity having;
  • TrmD having an amino acid sequence identical to that shown in SEQ ID NO. 64 has at least 86% identity and more preferably at least 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid trmU encoding a methyltransferase (methyltransferase, E.C. 2.1.1.61) having a nucleotide sequence corresponding to that shown in SEQ ID NO. At least 83% identity, and more preferably at least 85%, 87.5%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • TrmU having an amino acid sequence identical to that shown in SEQ ID NO. 66 has at least 92% identity and more preferably at least 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid yycF coding for a putative two-component response regulator with a nucleotide sequence the to SEQ ID NO. 67 has at least 83% identity and more preferably at least 85%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • a putative two-component response regulator YycF having an amino acid sequence identical to that shown in SEQ ID NO. At least 93% identity, and more preferably at least 94%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • a yycG nucleic acid encoding a putative two-component response regulator with a nucleotide sequence corresponding to that shown in SEQ ID NO. 69 has at least 78% identity and more preferably at least 80%, 85%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • a putative two-component response regulator YycG having an amino acid sequence identical to that shown in SEQ ID NO. At least 78% identity and more preferably at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid nusA encoding a putative transcription termination factor having a nucleotide sequence corresponding to that shown in SEQ ID NO. 71 has at least 84% identity and more preferably at least 85%, 87.5%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • a putative transcription termination factor NusA having an amino acid sequence identical to that shown in SEQ ID NO. 72 has at least 92% identity and more preferably at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid rplA coding for a 50S ribosomal protein L1 (5OS ribosomal protein L1, BL1), having a nucleotide sequence which corresponds to that shown in SEQ ID NO. 73 nucleotide sequence has at least 89% identity and increasingly preferably at least 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and particularly preferably 100% identity;
  • a 50S ribosomal protein L1 (5OS ribosomal protein L1; BL1) RpIA having an amino acid sequence corresponding to that shown in SEQ ID NO. 74 indicated amino acid sequence at least 91% identity, and more preferably at least 92.5%, 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity;
  • nucleic acid rplB coding for a 50S ribosomal protein L2 (5OS ribosomal protein L2), having a nucleotide sequence which corresponds to that shown in SEQ ID NO. 75 nucleotide sequence has at least 94% identity and increasingly preferably at least 95%, 96%, 97%, 98%, 99% and particularly preferably 100% identity;
  • 50S ribosomal protein L2 (5OS ribosomal protein L2) RpIB having an amino acid sequence corresponding to that shown in SEQ ID NO. At least 95% identity, and more preferably at least 96%, 96.5%, 97%, 97.5%, 98%, 98.5%, 99%, 99.5%, and most preferably 100% identity;
  • nucleic acid rpIC coding for a 50S ribosomal protein L3 (5OS ribosomal protein L3) having a nucleotide sequence which corresponds to that shown in SEQ ID NO. 77 nucleotide sequence has at least 93% identity, and more preferably at least 94%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • a 50S ribosomal protein L3 (5OS ribosomal protein L3) RpIC having an amino acid sequence identical to that shown in SEQ ID NO. 78 has at least 94% identity and, more preferably, at least 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity;
  • nucleic acid rpID encoding a 50S ribosomal protein L4 (5OS ribosomal protein L4) having a nucleotide sequence corresponding to that shown in SEQ ID NO. 79 nucleotide sequence at least 94% identity and increasingly preferably at least 95%, 96%, 96.5%, 97%, 97.5%, 98%, 98.5%, 99% 99.5% and particularly preferably 100% identity having;
  • a 50S ribosomal protein L4 (5OS ribosomal protein L4) RpID, having an amino acid sequence identical to that shown in SEQ ID NO. 80 has at least 93% identity and more preferably at least 94%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid rplE coding for a 50S ribosomal protein L5 (5OS ribosomal protein L5), having a nucleotide sequence which corresponds to that shown in SEQ ID NO. 81 nucleotide sequence has at least 94% identity and increasingly preferably at least 95%, 96%, 97%, 98%, 98.5%, 99%, 99.5% and most preferably 100% identity;
  • a 50S ribosomal protein L5 (5OS ribosomal protein L5) RpIE having an amino acid sequence corresponding to that shown in SEQ ID NO. 82 has at least 94% identity and more preferably at least 95%, 96%, 97%, 98%, 98.5%, 99%, 99.5% and most preferably 100% identity;
  • a nucleic acid rplF encoding a 50S ribosomal protein L6 (5OS ribosomal protein L6), having a nucleotide sequence corresponding to that shown in SEQ ID NO. 83 has at least 92% identity and more preferably at least 93%, 94%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • a 50S ribosomal protein L6 (5OS ribosomal protein L6) RpIF having an amino acid sequence corresponding to that shown in SEQ ID NO. 84 has at least 92% identity and more preferably at least 93%, 94%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid rpIL coding for a ribosomal protein of the large subunit L9P having a nucleotide sequence which corresponds to that shown in SEQ ID NO. 85 nucleotide sequence has at least 75% identity and increasingly preferably at least 80%, 85%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • a ribosomal protein of the large subunit L9P (LSU ribosomal protein L9P) RpIL having an amino acid sequence corresponding to that shown in SEQ ID NO. 86 has at least 84% identity and more preferably at least 85%, 90%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid rplJ coding for a ribosomal protein of the large subunit L10P having a nucleotide sequence which corresponds to that shown in SEQ ID NO. 87 nucleotide sequence has at least 95% identity, and more preferably at least 96%, 97%, 97.5%, 98%, 98.5%, 99%, 99.5% and most preferably 100% identity;
  • a ribosomal protein of the large subunit L10P (LSU ribosomal protein L10P) RpIJ, having an amino acid sequence corresponding to that shown in SEQ ID NO. At least 94% identity, and more preferably at least 95%, 96%, 97%, 97.5%, 98%, 98.5%, 99%, 99.5%, and most preferably 100% identity;
  • nucleic acid rpIL coding for a ribosomal protein of the large subunit L12P having a nucleotide sequence which corresponds to that shown in SEQ ID NO. 89 nucleotide sequence has at least 96% identity and more preferably at least 97%, 97.5%, 98%, 98.5%, 99%, 99.5% and most preferably 100% identity;
  • a ribosomal protein of the large subunit L12P (LSU ribosomal protein L12P; L7 / L12) RpIL having an amino acid sequence corresponding to that shown in SEQ ID NO. 90 indicated Amino acid sequence having at least 96% identity, and more preferably at least 97%, 97.5%, 98%, 98.5%, 99%, 99.5%, and most preferably 100% identity;
  • a nucleic acid rplM coding for a 50S ribosomal protein L13 having a nucleotide sequence which corresponds to that shown in SEQ ID NO. 91 has at least 97% identity and more preferably at least 97.5%, 98%, 98.5%, 99%, 99.5% and most preferably 100% identity
  • a 50S ribosomal protein L13 (5OS ribosomal protein L13) RpIM having an amino acid sequence corresponding to that shown in SEQ ID NO. 92 has at least 94% identity and more preferably at least 95%, 96%, 97%, 98%, 98.5%, 99%, 99.5% and most preferably 100% identity;
  • nucleic acid rplN coding for a 50S ribosomal protein L14 (5OS ribosomal protein L14), having a nucleotide sequence which corresponds to that shown in SEQ ID NO. 93 nucleotide sequence has at least 92% identity and more preferably at least 93%, 94%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • a 50S ribosomal protein L14 (5OS ribosomal protein L14) RpIN having an amino acid sequence corresponding to that shown in SEQ ID NO. 94 has at least 93% identity and more preferably at least 94%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid rplO coding for a 50S ribosomal protein L15 (5OS ribosomal protein L15), having a nucleotide sequence which corresponds to that shown in SEQ ID NO. 95 nucleotide sequence has at least 93% identity and increasingly preferably at least 94%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • a 50S ribosomal protein L15 (5OS ribosomal protein L15) RpIO, having an amino acid sequence corresponding to that shown in SEQ ID NO. At least 93% identity, and more preferably at least 94%, 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity of the indicated amino acid sequence;
  • nucleic acid rplP coding for a 50S ribosomal protein L16 (5OS ribosomal protein L16), having a nucleotide sequence which corresponds to that shown in SEQ ID NO. 97 nucleotide sequence has at least 94% identity and increasingly preferably at least 95%, 96%, 97%, 98%, 98.5%, 99%, 99.5% and most preferably 100% identity;
  • a 50S ribosomal protein L16 (5OS ribosomal protein L16) RpIP having an amino acid sequence corresponding to that shown in SEQ ID NO. 98 has at least 96% identity, and more preferably at least 96%, 97%, 98%, 98.5%, 99%, 99.5%, and most preferably 100% identity;
  • a 50S ribosomal protein L17 (5OS ribosomal protein L17) RpIQ having an amino acid sequence corresponding to that shown in SEQ ID NO. At least 94% identity, and more preferably at least 95%, 96%, 97%, 98%, 98.5%, 99%, 99.5% and most preferably 100% identity of the indicated amino acid sequence;
  • nucleic acid rpIR encoding a 50S ribosomal protein L18 (5OS ribosomal protein L18) having a nucleotide sequence corresponding to that shown in SEQ ID NO. 101 nucleotide sequence has at least 85% identity and more preferably at least 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • a 50S ribosomal protein L18 (5OS ribosomal protein L18) RpIR having an amino acid sequence corresponding to that shown in SEQ ID NO. 102 has at least 87% identity and more preferably at least 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid rpIS coding for a 50S ribosomal protein L19 having a nucleotide sequence which corresponds to that shown in SEQ ID NO. 103 nucleotide sequence has at least 91% identity, and more preferably at least 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • a 50S ribosomal protein L19 (5OS ribosomal protein L19) RpIS, having an amino acid sequence identical to that shown in SEQ ID NO. At least 93% identity, and more preferably at least 94%, 95%, 96%, 97%, 98%, 98.5%, 99%, 99.5%, and most preferably 100% identity;
  • nucleic acid rpIT coding for a ribosomal protein of the large subunit L20P having a nucleotide sequence which corresponds to that shown in SEQ ID NO. At least 93% identity, and more preferably at least 94%, 95%, 96%, 97%, 98%, 98.5%, 99%, 99.5%, and most preferably 100% identity of the indicated nucleotide sequence;
  • a ribosomal protein of the large subunit L20P (LSU ribosomal protein L20P) RpIT having an amino acid sequence identical to that shown in SEQ ID NO. 106 has at least 98% identity and, more preferably, at least 98%, 98.5%, 99%, 99.5%, and most preferably 100% identity;
  • nucleic acid coding for a 50S ribosomal protein L21 (5OS ribosomal protein L21), having a nucleotide sequence which corresponds to that shown in SEQ ID NO. 107 indicated Nucleotide sequence having at least 88% identity, and more preferably at least 90%, 92.5%, 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity;
  • a 50S ribosomal protein L21 (5OS ribosomal protein L21) RpIU with an amino acid sequence corresponding to that shown in SEQ ID NO. Has at least 86% identity, and more preferably at least 88%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity;
  • nucleic acid rplV coding for a 50S ribosomal protein L22 (5OS ribosomal protein L22), having a nucleotide sequence which corresponds to that shown in SEQ ID NO. At least 93% identity, and more preferably at least 94%, 95%, 96%, 97%, 98%, 98.5%, 99%, 99.5%, and most preferably 100% identity of the indicated nucleotide sequence;
  • a 50S ribosomal protein L22 (5OS ribosomal protein L22) RpIV having an amino acid sequence corresponding to that shown in SEQ ID NO. At least 93% identity, and more preferably at least 94%, 95%, 96%, 97%, 98%, 98.5%, 99%, 99.5% and most preferably 100% identity;
  • nucleic acid rplW coding for a 50S ribosomal protein L23 (5OS ribosomal protein L23), having a nucleotide sequence which corresponds to that shown in SEQ ID NO. At least 93% identity, and more preferably at least 94%, 95%, 96%, 97%, 98%, 98.5%, 99%, 99.5% and most preferably 100% identity;
  • a 50S ribosomal protein L23 (5OS ribosomal protein L23) RpIW, having an amino acid sequence corresponding to that shown in SEQ ID NO. 112 has at least 88% identity and more preferably at least 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid rpIX coding for a 50S ribosomal protein L24 (5OS ribosomal protein L24), having a nucleotide sequence which corresponds to that shown in SEQ ID NO. 113 nucleotide sequence has at least 92% identity and increasingly preferably at least 93%, 94%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • a 50S ribosomal protein L24 (50S ribosomal protein L24) RpIX having an amino acid sequence identical to that shown in SEQ ID NO. 114 amino acid sequence has at least 94% identity and increasingly preferably at least 95%, 96%, 97%, 98%, 98.5%, 99%, 99.5% and most preferably 100% identity;
  • nucleic acid rpmA encoding a 50S ribosomal protein L27 (5OS ribosomal protein L27) having a nucleotide sequence corresponding to that shown in SEQ ID NO. 115 indicated Nucleotide sequence having at least 94% identity, and more preferably at least 95%, 96%, 97%, 98%, 98.5%, 99%, 99.5%, and most preferably 100% identity;
  • a 50S ribosomal protein L27 (5OS ribosomal protein L27) RpmA having an amino acid sequence corresponding to that shown in SEQ ID NO. At least 93% identity, and more preferably at least 94%, 95%, 96%, 97%, 98%, 98.5%, 99%, 99.5%, and most preferably 100% identity;
  • a 50S ribosomal protein L28 (5OS ribosomal protein L28) RpmB having an amino acid sequence corresponding to that shown in SEQ ID NO. 118 at least 92% identity and increasingly preferably at least 93%, 94%, 95%, 96%, 97%, 98%, 99% and particularly preferably 100% identity.
  • nucleic acid rpmC encoding a 50S ribosomal protein L29 (5OS ribosomal protein L29), having a nucleotide sequence corresponding to that shown in SEQ ID NO. 119 nucleotide sequence has at least 96% identity and more preferably at least 97%, 97.5%, 98%, 98.5%, 99%, 99.5% and most preferably 100% identity;
  • a 50S ribosomal protein L29 (5OS ribosomal protein L29) RpmC having an amino acid sequence corresponding to that shown in SEQ ID NO. 120 amino acid sequence has at least 99% identity and increasingly preferably at least 99.5% and particularly preferably 100% identity;
  • nucleic acid rpmD coding for a 50S ribosomal protein L30 (5OS ribosomal protein L30), having a nucleotide sequence which corresponds to that shown in SEQ ID NO. 121 nucleotide sequence has at least 93% identity and increasingly preferably at least 94%, 95%, 96%, 97%, 98%, 98.5%, 99%, 99.5% and most preferably 100% identity;
  • a 50S ribosomal protein L30 (5OS ribosomal protein L30) RpmD having an amino acid sequence identical to that shown in SEQ ID NO. 122 has at least 99% identity and more preferably at least 99.5% and most preferably 100% identity;
  • nucleic acid rpmE coding for a ribosomal protein L31 (ribosomal protein L31), having a nucleotide sequence which corresponds to that shown in SEQ ID NO. 123 nucleotide sequence has at least 95% identity and increasingly preferably at least 96%, 97%, 97.5%, 98%, 98.5%, 99%, 99.5% and most preferably 100% identity;
  • a ribosomal protein L31 (ribosomal protein L31) RpmE having an amino acid sequence corresponding to that shown in SEQ ID NO. 124 has at least 99% identity and more preferably at least 99.5% and most preferably 100% identity;
  • nucleic acid rpmF encoding a 50S ribosomal protein L32 (5OS ribosomal protein L32) having a nucleotide sequence corresponding to that shown in SEQ ID NO. 125 nucleotide sequence has at least 83% identity, and more preferably at least 85%, 87.5%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • a 50S ribosomal protein L32 (5OS ribosomal protein L32) RpmF, having an amino acid sequence identical to that shown in SEQ ID NO. Has at least 71% identity, and more preferably at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity;
  • nucleic acid rpmGA coding for a type 1-50S ribosomal protein L33 (5OS ribosomal protein L33 type 1), having a nucleotide sequence which corresponds to that shown in SEQ ID NO. 127 nucleotide sequence has at least 90% identity and increasingly preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • a type 1-50S ribosomal protein L33 (5OS ribosomal protein L33 type 1) RpmGA having an amino acid sequence identical to that shown in SEQ ID NO. 128 has at least 94% identity and more preferably at least 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid rpmGB coding for a ribosomal protein of the large subunit L33P having a nucleotide sequence which corresponds to that shown in SEQ ID NO. 129 nucleotide sequence has at least 84% identity, and more preferably at least 85%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • L33P L33P ribosomal protein L33P
  • RpmGB having an amino acid sequence corresponding to that shown in SEQ ID NO. 130 has at least 94% identity and more preferably at least 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid rpml encoding a 50S ribosomal protein L35 (5OS ribosomal protein L35), having a nucleotide sequence corresponding to that shown in SEQ ID NO. 131 nucleotide sequence at least 88% identity and increasingly preferably at least 90%, 92.5%, 94%, 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity;
  • Amino acid sequence corresponding to that shown in SEQ ID NO. 132 indicated amino acid sequence at least 94% identity and increasingly preferably at least 95%, 96%, 97%, 98%,
  • nucleic acid rpmJ encoding a 50S ribosomal protein L36 (5OS ribosomal protein L36) having a nucleotide sequence corresponding to that shown in SEQ ID NO. 133 nucleotide sequence has at least 93% identity, and more preferably at least 94%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • a 50S ribosomal protein L36 (5OS ribosomal protein L36) RpmJ, having an amino acid sequence identical to that shown in SEQ ID NO. 134 amino acid sequence has at least 94% identity and increasingly preferably at least 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid rpsB coding for a ribosomal protein S2 (ribosomal protein S2) having a nucleotide sequence which corresponds to that shown in SEQ ID NO. 135 nucleotide sequence has at least 92% identity, and more preferably at least 93%, 94%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • Ribosomal Protein S2 Ribosomal Protein S2 (ribosomal protein S2) RpsB having an amino acid sequence corresponding to that shown in SEQ ID NO. 136 has at least 94% identity and, more preferably, at least 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity;
  • nucleic acid rpsC encoding a 30S ribosomal protein S3 (3OS ribosomal protein S3) having a nucleotide sequence corresponding to that shown in SEQ ID NO. 137 nucleotide sequence has at least 94% identity and increasingly preferably at least 95%, 96%, 97%, 98%, 98.5%, 99%, 99.5% and most preferably 100% identity;
  • a 30S ribosomal protein S3 (3OS ribosomal protein S3) RpsC having an amino acid sequence identical to that shown in SEQ ID NO. At least 94% identity, and more preferably at least 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid rpsD encoding a 30S ribosomal protein S4 (3OS ribosomal protein S4), having a nucleotide sequence corresponding to that shown in SEQ ID NO. 139 nucleotide sequence has at least 89% identity and increasingly preferably at least 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and particularly preferably 100% identity;
  • a 30S ribosomal protein S4 (3OS ribosomal protein S4) RpsD having an amino acid sequence corresponding to that shown in SEQ ID NO. 140 amino acid sequence has at least 94% identity and increasingly preferably at least 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid rpsE encoding a 30S ribosomal protein S5 (3OS ribosomal protein S5), having a nucleotide sequence corresponding to that shown in SEQ ID NO. 141 nucleotide sequence has at least 95% identity and increasingly preferably at least 96%, 97%, 98%, 98.5%, 99%, 99.5% and most preferably 100% identity;
  • a 30S ribosomal protein S5 (3OS ribosomal protein S5) RpsE, having an amino acid sequence identical to that shown in SEQ ID NO. 142 has at least 94% identity and more preferably at least 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid rpsF coding for a ribosomal protein S6 (ribosomal protein S6), having a nucleotide sequence which corresponds to that shown in SEQ ID NO. At least 91% identity, and more preferably at least 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • ribosomal protein S6 (ribosomal protein S6) RpsF having an amino acid sequence identical to that shown in SEQ ID NO. 144 has at least 92% identity and more preferably at least 93%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid rpsG coding for a ribosomal protein of the small subunit S7P having a nucleotide sequence which corresponds to that shown in SEQ ID NO. 145 nucleotide sequence has at least 95% identity and increasingly preferably at least 96%, 97%, 97.5%, 98%, 98.5%, 99%, 99.5% and most preferably 100% identity;
  • a ribosomal protein of the small subunit S7P (SSU ribosomal protein S7P) RpsG having an amino acid sequence corresponding to that shown in SEQ ID NO. 146 has at least 94% identity and more preferably at least 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid rpsH encoding a 30S ribosomal protein S8 (3OS ribosomal protein S8) having a nucleotide sequence corresponding to that shown in SEQ ID NO. At least 91% identity, and more preferably at least 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • a 30S ribosomal protein S8 (3OS ribosomal protein S8) RpsH having an amino acid sequence identical to that shown in SEQ ID NO. 148 indicated amino acid sequence at least 94% identity, and more preferably at least 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid rps1 encoding a 30S ribosomal protein S9 (3OS ribosomal protein S9) having a nucleotide sequence corresponding to that shown in SEQ ID NO. 149 nucleotide sequence having at least 95% identity and more preferably at least 96%, 97%, 97.5%, 98%, 98.5%, 99%, 99.5% and most preferably 100% identity;
  • a 30S ribosomal protein S9 (3OS ribosomal protein S9) Rps1, having an amino acid sequence identical to that shown in SEQ ID NO. 150 has at least 94% identity and more preferably at least 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • a nucleic acid rpsJ coding for a ribosomal protein S10 (ribosomal protein S10) having a nucleotide sequence which corresponds to that shown in SEQ ID NO. At least 97% identity and more preferably at least 97.5%, 98%, 98.5%, 99%, 99.5% and most preferably 100% identity
  • a ribosomal protein S10 (ribosomal protein S10) RpsJ having an amino acid sequence identical to that shown in SEQ ID NO. 152 has at least 94% identity and more preferably at least 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid rpsK coding for a 30S-ribosomal protein S11 (3OS ribosomal protein S11), having a nucleotide sequence which corresponds to that shown in SEQ ID NO. At least 93% identity, and more preferably at least 94%, 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity, indicated nucleotide sequence;
  • a 30S ribosomal protein S11 (3OS ribosomal protein S11) RpsK having an amino acid sequence corresponding to that shown in SEQ ID NO. 154 has at least 94% identity and, more preferably, at least 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity;
  • nucleic acid rpsL encoding a small subunit ribosomal protein S12P (SSU ribosomal protein S12P) having a nucleotide sequence corresponding to that shown in SEQ ID NO. At least 93% identity, and more preferably at least 94%, 95%, 96%, 97%, 98%, 99%, 99.5%, and most preferably 100% identity of the indicated nucleotide sequence;
  • a small subunit S12P (SSU ribosomal protein S12P) ribosomal protein RpsL having an amino acid sequence corresponding to that shown in SEQ ID NO. 156 has at least 94% identity and more preferably at least 95%, 96%, 97%, 98%, 99% and most preferably 100% identity
  • a nucleic acid rpsM encoding a 30S ribosomal protein S13 (3OS ribosomal protein S13), having a nucleotide sequence corresponding to that shown in SEQ ID NO. 157 has at least 94% identity and more preferably at least 95%, 96%, 97%, 97.5%, 98%, 98.5%, 99%, 99.5% and most preferably 100% identity;
  • a 30S ribosomal protein S13 (3OS ribosomal protein S13) RpsM, having an amino acid sequence identical to that shown in SEQ ID NO. 158 has at least 94% identity and more preferably at least 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid rpsN encoding a 30S ribosomal protein S14-1 (3OS ribosomal protein S14-1), having a nucleotide sequence corresponding to that shown in SEQ ID NO. At least 93% identity, and more preferably at least 94%, 95%, 96%, 97%, 98%, 98.5%, 99%, 99.5% and most preferably 100% identity;
  • a 30S ribosomal protein S14-1 (3OS ribosomal protein S14-1) RpsN having an amino acid sequence corresponding to that shown in SEQ ID NO. 160 has at least 94% identity and more preferably at least 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid rpsO coding for a ribosomal protein S15 (ribosomal protein S15), having a nucleotide sequence which corresponds to that shown in SEQ ID NO. At least 93% identity, and more preferably at least 94%, 95%, 96%, 97%, 98%, 98.5%, 99%, 99.5%, and most preferably 100% identity;
  • a ribosomal protein S15 (ribosomal protein S15) RpsO having an amino acid sequence corresponding to that shown in SEQ ID NO. 162 has at least 94% identity and more preferably at least 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid rpsP encoding a 30S ribosomal protein S16 (3OS ribosomal protein S16), having a nucleotide sequence corresponding to that shown in SEQ ID NO. At least 93% identity, and more preferably at least 94%, 95%, 96%, 97%, 98%, 98.5%, 99%, 99.5% and most preferably 100% identity;
  • a 30S ribosomal protein S16 (3OS ribosomal protein S16) RpsP having an amino acid sequence corresponding to that shown in SEQ ID NO. 164 indicated amino acid sequence at least 94% identity, and more preferably at least 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid rpsQ coding for a 30S-ribosomal protein S17 (3OS ribosomal protein S 17), having a nucleotide sequence which corresponds to that shown in SEQ ID NO. 165 nucleotide sequence has at least 93% identity and increasingly preferably at least 94%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • a 30S ribosomal protein S17 (3OS ribosomal protein S17) RpsQ, having an amino acid sequence identical to that shown in SEQ ID NO. 166 has at least 94% identity and more preferably at least 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • a nucleic acid rpsR encoding a ribosomal protein S18 (ribosomal protein S18) having a nucleotide sequence corresponding to that shown in SEQ ID NO. 167 has at least 94% identity and more preferably at least 95%, 96%, 97%, 98%, 98.5%, 99%, 99.5% and most preferably 100% identity;
  • a ribosomal protein S18 (ribosomal protein S18) RpsR having an amino acid sequence identical to that shown in SEQ ID NO. 168 has at least 94% identity and more preferably at least 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid rpsS coding for a 30S ribosomal protein S19 (3OS ribosomal protein S19), having a nucleotide sequence which corresponds to that shown in SEQ ID NO. At least 93% identity, and more preferably at least 94%, 95%, 96%, 97%, 98%, 98.5%, 99%, 99.5% and most preferably 100% identity;
  • a 30S ribosomal protein S19 (3OS ribosomal protein S 19) RpsS having an amino acid sequence corresponding to that shown in SEQ ID NO. 170 has at least 94% identity and more preferably at least 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid rpsT encoding a 30S ribosomal protein S20 (3OS ribosomal protein S20), having a nucleotide sequence corresponding to that shown in SEQ ID NO. At least 87% identity, and more preferably at least 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • a 30S ribosomal protein S20 (3OS ribosomal protein S20) RpsT having an amino acid sequence identical to that shown in SEQ ID NO. 172 has at least 94% identity and more preferably at least 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • a nucleic acid rpsU encoding a small subunit ribosomal protein S21P (SSU ribosomal protein S21P) having a nucleotide sequence corresponding to that shown in SEQ ID NO. At least 96% identity and more preferably at least 96.5%, 97%, 97.5%, 98%, 98.5%, 99%, 99.5%, and most preferably 100% identity;
  • a small subunit S21 P (SSU ribosomal protein S21 P) ribosomal protein RpsU, having an amino acid sequence corresponding to that shown in SEQ ID NO. 174 has at least 94% identity and, more preferably, at least 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity;
  • nucleic acid alaS coding for an alanyl-tRNA synthetase (alanyl-tRNA synthetase, E.C. 6.1.1.7), having a nucleotide sequence which corresponds to that shown in SEQ ID NO. At least 81% identity, and more preferably at least 82.5%, 85%, 87.5%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99%, and most preferably 100, of nucleotide sequence indicated % Identity;
  • AIaS having an amino acid sequence corresponding to that shown in SEQ ID NO. At least 84% identity, and more preferably at least 85%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity of the indicated amino acid sequence;
  • ArgS probable with an amino acid sequence corresponding to that shown in SEQ ID NO. 178 has at least 92% identity and more preferably at least 93%, 94%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • an asnS nucleic acid encoding an asparaginyl-tRNA synthetase (asparaginyl-tRNA synthetase) having a nucleotide sequence corresponding to that shown in SEQ ID NO. At least 86% identity, and more preferably at least 87.5%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99%, and most preferably
  • an asparaginyl-tRNA synthetase (asparaginyl-tRNA synthetase) AsnS having an amino acid sequence corresponding to that shown in SEQ ID NO. 180 indicated amino acid sequence at least 96% identity, and more preferably at least 97%, 98%, 99%, and most preferably 100% identity;
  • nucleic acid aspS coding for an aspartyl-tRNA synthetase (aspartyl-tRNA synthetase; E.C. 6.1.1.12), having a nucleotide sequence which corresponds to that shown in SEQ ID NO. At least 80% identity, and more preferably at least 82.5%, 85%, 87.5%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99%, and most preferably 100, nucleotide sequence indicated % Identity;
  • an aspartyl-tRNA synthetase (aspartyl-tRNA synthetase; E.C. 6.1.1.12) AspS, having an amino acid sequence identical to that shown in SEQ ID NO. 182 has at least 85% identity, and more preferably at least 87.5%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity;
  • nucleic acid cysS coding for a cysteinyl-tRNA synthetase having a nucleotide sequence which corresponds to that shown in SEQ ID NO. At least 80% identity, and more preferably at least 85%, 87.5%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • CysS having an amino acid sequence corresponding to that shown in SEQ ID NO. 184 has at least 87% identity and more preferably at least 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • a nucleic acid gltX encoding a glutamyl-tRNA synthetase (glutamyl-tRNA synthetase) having a nucleotide sequence corresponding to that shown in SEQ ID NO. 185 nucleotide sequence has at least 82% identity and increasingly preferably at least 85%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and particularly preferably 100% identity;
  • a glutamyl-tRNA synthetase (glutamyl-tRNA synthetase) GItX having an amino acid sequence identical to that shown in SEQ ID NO. 186 has at least 90% identity, and more preferably at least 92%, 94%, 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity;
  • nucleic acid glyQ coding for an alpha-chain of a glycyl-tRNA synthetase (glycyl-tRNA synthetase alpha chain, EC 6.1.1.14), having a nucleotide sequence which corresponds to that shown in SEQ ID NO.
  • an alpha-chain of a glycyl-tRNA synthetase (glycyl-tRNA synthetase alpha chain; EC 6.1.1.14)
  • GIyQ having an amino acid sequence which corresponds to the amino acid sequence shown in SEQ ID NO. 188 has at least 96% identity and more preferably at least 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid glyS coding for a beta-chain of a glycyl-tRNA synthetase (glycyl-tRNA synthetase beta chain, E.C. 6.1.1.14), having a nucleotide sequence which corresponds to that shown in SEQ ID NO. At least 74% identity, and more preferably at least 75%, 80%, 85%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • GIyS a beta-chain of a glycyl-tRNA synthetase (glycyl-tRNA synthetase beta chain; E.C. 6.1.1.14) GIyS, having an amino acid sequence identical to that shown in SEQ ID NO. At least 76% identity, and more preferably at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity of the indicated amino acid sequence;
  • nucleic acid hisS coding for a histidyl-tRNA synthetase having a nucleotide sequence which corresponds to that shown in SEQ ID NO. 191 has at least 81% identity, and more preferably at least 85%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity;
  • HisS having an amino acid sequence identical to that shown in SEQ ID NO. 192 having at least 85% identity and more preferably at least 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid HeS coding for an isoleucyl-tRNA synthetase (isoleucyl-tRNA synthetase, E.C. 6.1.1.5), having a nucleotide sequence which corresponds to that shown in SEQ ID NO. 193 nucleotide sequence has at least 80% identity and more preferably at least 85%, 90%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • NeS having an amino acid sequence corresponding to that shown in SEQ ID NO. 194 has at least 85% identity, and more preferably at least 87.5%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity;
  • leuS nucleic acid encoding a leucyl-tRNA synthetase (leucyl-tRNA synthetase; EC 6.1.1.4) having a nucleotide sequence corresponding to that shown in SEQ ID NO. 195 nucleotide sequence at least 84% identity and increasingly preferred at least 85%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • Leucyl tRNA synthetase leucyl tRNA synthetase; E.C. 6.1.1.4
  • LeuS having an amino acid sequence which corresponds to the amino acid sequence shown in SEQ ID NO. Has at least 90% identity, and more preferably at least 92.5%, 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity;
  • a lysS nucleic acid encoding a lysyl tRNA synthetase (lysyl-tRNA synthetase) having a nucleotide sequence corresponding to that shown in SEQ ID NO. At least 83% identity, and more preferably at least 85%, 90%, 95%, 96%, 96.5%, 97%, 97.5%, 98%, 98.5%, 99% 99.5%, of the nucleotide sequence indicated. and most preferably 100% identity;
  • lysyl-tRNA synthetase (lysyl-tRNA synthetase) LysS, having an amino acid sequence identical to that shown in SEQ ID NO. At least 92% identity and more preferably at least 94%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid metS coding for a methionyl-tRNA synthetase (methionyl-tRNA synthetase, E.C. 6.1.1.10), having a nucleotide sequence which corresponds to that shown in SEQ ID NO. At least 80% identity, and more preferably at least 85%, 90%, 95%, 96%, 97%, 98%, 98.5%, 99%, 99.5%, and most preferably 100% identity of the indicated nucleotide sequence;
  • methionyl-tRNA synthetase (methionyl-tRNA synthetase; E.C. 6.1.1.10) MetS having an amino acid sequence identical to that shown in SEQ ID NO. Having at least 89% identity, and more preferably at least 90%, 92.5%, 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity of the indicated amino acid sequence;
  • nucleic acid pheS coding for an alpha-chain of a phenylalanyl-tRNA synthetase (phenylalanyl-tRNA synthetase, alpha chain; E.C. 6.1.1.20), having a nucleotide sequence which corresponds to that shown in SEQ ID NO. 201 has at least 82% identity and more preferably at least 85%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • 202 has at least 94% identity and more preferably at least 95%, 96%, 97%, 98%, 99% and most preferably 100% identity; a nucleic acid pheT coding for a phenylalanyl-tRNA synthetase ⁇ -chain (phenylalanyl-tRNA synthetase, beta chain; EC 6.1.1.20), having a nucleotide sequence which corresponds to that shown in SEQ ID NO. At least 77% identity and more preferably at least 80%, 85%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • a beta-chain of phenylalanyl-tRNA synthetase (phenylalanyl-tRNA synthetase, beta chain; E.C. 6.1.1.20)
  • PheT having an amino acid sequence corresponding to that shown in SEQ ID NO. 204 has at least 82% identity, and more preferably at least 85%, 90%, 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity;
  • nucleic acid proS coding for a prolyl-tRNA synthetase (prolyl-tRNA synthetase, E.C. 6.1.1.15), having a nucleotide sequence which corresponds to that shown in SEQ ID NO. 205 nucleotide sequence at least 77% identity and increasingly preferably at least 80%, 85%, 90%, 95%, 96%, 97%, 97.5%, 98%, 98.5%, 99%, 99.5% and most preferably 100% identity;
  • prolyl-tRNA synthetase having an amino acid sequence identical to that shown in SEQ ID NO. 206 has at least 84% identity, and more preferably at least 87.5%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid serS coding for a seryl-tRNA synthetase (seryl-tRNA synthetase, E.C. 6.1.1.11), having a nucleotide sequence which corresponds to that shown in SEQ ID NO. At least 85% identity, and more preferably at least 90%, 95%, 97%, 97.5%, 98%, 98.5%, 99%, 99.5%, and most preferably 100% identity;
  • seryl-tRNA synthetase a seryl-tRNA synthetase (seryl-tRNA synthetase; E.C. 6.1.1.11)
  • SerS having an amino acid sequence corresponding to that shown in SEQ ID NO. At least 95% identity, and more preferably at least 97%, 97.5%, 98%, 98.5%, 99%, 99.5% and most preferably 100% identity;
  • a nucleic acid trpS coding for a tryptophanyl-tRNA synthetase (tryptophanyl-tRNA synthetase, E.C. 6.1.1.2), having a nucleotide sequence which corresponds to that shown in SEQ ID NO. At least 83% identity, and more preferably at least 85%, 87.5%, 90%, 92.5%, 95%, 97.5%, 98%, 99% and most preferably 100% identity;
  • TrpS having an amino acid sequence corresponding to that shown in SEQ ID NO. 210 indicated Amino acid sequence having at least 94% identity, and more preferably at least 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity;
  • a tyrS nucleic acid encoding a tyrosyl-tRNA synthetase (tyrosyl-tRNA synthetase) having a nucleotide sequence corresponding to that shown in SEQ ID NO. 211 has at least 83% identity and more preferably at least 85%, 90%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • a tyrosyl-tRNA synthetase (tyrosyl-tRNA synthetase) TyrS having an amino acid sequence identical to that shown in SEQ ID NO. 212 has at least 94% identity and more preferably at least 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • a nucleic acid valS coding for a valyl-tRNA synthetase (valyl-tRNA synthetase, E.C. 6.1.1.9), having a nucleotide sequence which corresponds to that shown in SEQ ID NO. 213 has at least 84% identity, and more preferably at least 85%, 90%, 92.5%, 94%, 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity;
  • valyl-tRNA synthetase having an amino acid sequence identical to that shown in SEQ ID NO. 214 has at least 94% identity and more preferably at least 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid gatA coding for a Gln / glutamyl-tRNA (Gln) amidotransferase (glutamyl-tRNA (Gln) amidotransferase; E.C. 6.3.5.-), having a nucleotide sequence which corresponds to that shown in SEQ ID NO. 215 has at least 86% identity, and more preferably at least 90%, 95%, 96%, 97%, 98%, 98.5%, 99%, 99.5%, and most preferably 100% identity;
  • GatA having an amino acid sequence corresponding to that shown in SEQ ID NO. Having at least 94% identity, and more preferably at least 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity;
  • a gatB nucleic acid encoding a B subunit of a glutamyl-tRNA (Gln) amidotransferase (glutamyl-tRNA (Gln) amidotransferase subunit B, EC 6.3.5.-) having a nucleotide sequence corresponding to that shown in SEQ ID NO , 217 has at least 89% identity and more preferably at least 90%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity; a B subunit of a glutamyl-tRNA (Gln) amidotransferase (glutamyl-tRNA (Gln) amidotransferase subunit B; EC 6.3.5.-) GatB having an amino acid sequence identical to that shown in SEQ ID NO. 218 has at least 94% identity and more preferably at least 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid gatC coding for a subunit C of a glutamyl-tRNA (Gln) amidotransferase (glutamyl-tRNA (Gln) amidotransferase subunit C, E.C. 6.3.5.-), having a nucleotide sequence which corresponds to that shown in SEQ ID NO. 219 has at least 83% identity and more preferably at least 85%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • GatC having an amino acid sequence identical to that shown in SEQ ID NO. 220 has at least 94% identity and more preferably at least 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid encoding a methionyl-tRNA formyltransferase (methionyl-tRNA formyltransferase, E.C. 2.1.2.9) having a nucleotide sequence corresponding to that shown in SEQ ID NO. At least 77% identity and, more preferably, at least 80%, 85%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity are given ;
  • Fmt having an amino acid sequence identical to that shown in SEQ ID NO. 222 has at least 94% identity and, more preferably, at least 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity;
  • nucleic acid frr coding for a ribosome recycling factor having a nucleotide sequence corresponding to that shown in SEQ ID NO. At least 82% identity, and more preferably at least 85%, 90%, 92%, 94%, 95%, 96%, 97%, 98%, 98.5%, 99%, 99.5%, and most preferably, of nucleotide sequence indicated 100% identity;
  • a Ribosome Recycling Factor Frr having an amino acid sequence corresponding to that shown in SEQ ID NO. 224 has at least 94% identity and more preferably at least 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid fusA coding for an elongation factor G of protein translation having a nucleotide sequence identical to that described in SEQ ID NO. 225 nucleotide sequence has at least 90% identity and more preferably at least 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • an elongation factor G of protein translation (protein translation elongation factor G; EF-G) FusA having an amino acid sequence corresponding to that shown in SEQ ID NO. At least 93% identity, and more preferably at least 94%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • a nucleic acid infA encoding a translation initiation factor IF-1 (translation initiation factor IF-1) having a nucleotide sequence corresponding to that shown in SEQ ID NO. At least 96% identity, and more preferably at least 96.5%, 97%, 97.5%, 98%, 98.5%, 99%, 99.5%, and most preferably 100% identity;
  • a translation initiation factor IF-1 (translation initiation factor IF-1) InfA having an amino acid sequence which corresponds to that shown in SEQ ID NO. 228 has at least 94% identity and, more preferably, at least 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity;
  • nucleic acid infB coding for a translation initiation factor IF-2 (translation initiation factor IF-2), having a nucleotide sequence corresponding to that shown in SEQ ID NO.
  • translation initiation factor IF-2 translation initiation factor IF-2
  • At least 84% identity and, more preferably, at least 90%, 92%, 94%, 95%, 96%, 97%, 98%, 98.5%, 99%, 99.5% and most preferably 100% identity are given having;
  • a translation initiation factor IF-2 (translation initiation factor IF-2) InfB, having an amino acid sequence identical to that shown in SEQ ID NO. 230 has at least 94% identity and more preferably at least 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid infC encoding a translation initiation factor IF-3, having a nucleotide sequence corresponding to that shown in SEQ ID NO. At least 73% identity and, more preferably, at least 75%, 80%, 85%, 90%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity are given ;
  • a translation initiation factor IF-3 translation initiation factor IF-3 (translation initiation factor IF-3) InfC having an amino acid sequence corresponding to that shown in SEQ ID NO. 232 has at least 94% identity and more preferably at least 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • a peptide chain release factor 1 PrfA having an amino acid sequence corresponding to that shown in SEQ ID NO. 234 has at least 90% identity and, more preferably, at least 92.5%, 94%, 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity;
  • a nucleic acid tsf coding for an elongation factor Ts (elongation factor Ts; EF-Ts), having a nucleotide sequence which corresponds to that shown in SEQ ID NO. 235 has at least 87% identity and more preferably at least 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • an elongation factor Ts (Elongation factor Ts; EF-Ts) Tsf having an amino acid sequence identical to that shown in SEQ ID NO. 236 has at least 94% identity and more preferably at least 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid tufA coding for a protein translation elongation factor Tu (Protein Translation Elongation Factor Tu; EF-TU) having a nucleotide sequence which corresponds to that shown in SEQ ID NO. 237 nucleotide sequence has at least 94% identity and more preferably at least 95%, 96%, 97%, 98%, 98.5%, 99%, 99.5% and most preferably 100% identity;
  • a protein translation elongation factor Tu (Protein Translation Elongation Factor Tu; EF-TU) TufA having an amino acid sequence identical to that shown in SEQ ID NO. 238 has at least 94% identity and more preferably at least 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • a nucleic acid spoVC encoding a peptidyl-tRNA-hydrolase (peptidyl-tRNA hydrolase) having a nucleotide sequence corresponding to that shown in SEQ ID NO. 239 has at least 81% identity, and more preferably at least 85%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity;
  • a peptidyl-tRNA-hydrolase (peptidyl-tRNA hydrolase) SpoVC having an amino acid sequence corresponding to that shown in SEQ ID NO. 240 has at least 94% identity and more preferably at least 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • a nucleic acid encoding a Class I heat shock protein (Class I heat shock protein; chaperonin) having a nucleotide sequence corresponding to that shown in SEQ ID NO. 241 has at least 91% identity, and more preferably at least 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity;
  • class I heat shock protein (Class I heat-shock protein; chaperonin) GroEL having an amino acid sequence corresponding to that shown in SEQ ID NO. 242 identity and, more preferably, at least 95%, 96%, 97%, 97.5%, 98%, 98.5%, 99%, 99.5% and most preferably 100% identity;
  • a nucleic acid groES encoding a Class I heat shock protein (Class I heat shock protein; chaperonin) having a nucleotide sequence corresponding to that shown in SEQ ID NO. 243 has at least 91% identity, and more preferably at least 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity;
  • a class I heat shock protein (Class I heat-shock protein; chaperonin) GroES having an amino acid sequence identical to that shown in SEQ ID NO. 244 has at least 98% identity and more preferably at least 98.5%, 99%, 99.5% and most preferably 100% identity;
  • nucleic acid map encoding a methionine aminopeptidase (methionine aminopeptidase, E.C., 3.4.11.18) having a nucleotide sequence corresponding to that shown in SEQ ID NO. 245 nucleotide sequence has at least 88% identity and more preferably at least 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • methionine aminopeptidase E.C., 3.4.11.18 map having an amino acid sequence identical to that shown in SEQ ID NO. 246 has at least 94% identity and more preferably at least 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • a nucleic acid ffh coding for a subunit FFH / SRP54 of a signal recognition particle (subunit FFH / SRP54) having a nucleotide sequence which corresponds to that shown in SEQ ID NO. 247 has at least 85% identity and more preferably at least 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • a subunit FFH / SRP54 of a signal recognition particle (subunit FFH / SRP54) Ffh having an amino acid sequence identical to that shown in SEQ ID NO. 248 has at least 94% identity and, more preferably, at least 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity;
  • nucleic acid ftsY coding for a signal recognition particle with a nucleotide sequence corresponding to that shown in SEQ ID NO. At least 78% identity and more preferably at least 80%, 85%, 87.5%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity are given having;
  • a Signal Recognition Particle FtsY having an amino acid sequence identical to that shown in SEQ ID NO. Having at least 82% identity, and more preferably at least 86%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity of the indicated amino acid sequence;
  • nucleic acid prsA coding for a precursor of a protein export protein precursor having a nucleotide sequence which corresponds to that shown in SEQ ID NO. At least 73% identity, and more preferably at least 75%, 80%, 85%, 87.5%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity;
  • PrsA having an amino acid sequence corresponding to that shown in SEQ ID NO. 252 has at least 94% identity and more preferably at least 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • a secE nucleic acid encoding a subunit of a preprotein translocase (subprotein) translocase having a nucleotide sequence corresponding to that shown in SEQ ID NO. At least 83% identity, and more preferably at least 85%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • Preprotein translocase subunit a subunit of a preproprotein translocase (Preprotein translocase subunit) SecE having an amino acid sequence which corresponds to the amino acid sequence shown in SEQ ID NO. At least 63% identity and more preferably at least 70%, 75%, 80%, 85%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% of the amino acid sequence indicated. Having identity;
  • nucleic acid secY coding for a subunit of a preprotein translocase preprotein translocase subunit
  • preprotein translocase subunit having a nucleotide sequence which corresponds to that shown in SEQ ID NO. 255 nucleotide sequence at least 81% identity and increasingly preferably at least 85%, 87.5%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • Preprotein translocase subunit a subunit of a preprotein translocase (Preprotein translocase subunit) SecY, having an amino acid sequence identical to that shown in SEQ ID NO. 256 amino acid sequence has at least 89% identity and more preferably at least 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid accA coding for an acetyl-CoA carboxylase having a nucleotide sequence which corresponds to that shown in SEQ ID NO. At least 80% identity, and more preferably at least 85%, 87.5%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • an acetyl-CoA carboxylase (acetyl-CoA carboxylase) AccA having an amino acid sequence identical to that shown in SEQ ID NO. 258 has at least 94% identity and more preferably at least 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid accB coding for a biotin carboxyl carrier protein of the acetyl-CoA carboxylase (biotin carboxyl carrier protein of acetyl-CoA carboxylase), having a nucleotide sequence which corresponds to that shown in SEQ ID NO. At least 76% identity, and more preferably at least 80%, 85%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • biotin-carboxyl carrier protein of the acetyl-CoA carboxylase biotin carboxyl carrier protein of acetyl-CoA carboxylase
  • AccB having an amino acid sequence which corresponds to the amino acid sequence shown in SEQ ID NO. 260 has at least 94% identity and more preferably at least 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid accC encoding a biotin carboxylase subunit of an acetyl-CoA carboxylase (acetyl-CoA carboxylase subunit; biotin carboxylase subunit; E.C. 6.4.1.2) having a nucleotide sequence corresponding to that shown in SEQ ID NO. At least 80% identity, and more preferably at least 85%, 87.5%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity;
  • biotin carboxylase subunit of an acetyl-CoA carboxylase acetyl-CoA carboxylase subunit; biotin carboxylase subunit; EC 6.4.1.2
  • AccC having an amino acid sequence identical to that shown in SEQ ID NO. 262 indicated amino acid sequence at least 94% identity, and more preferably at least 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid accD encoding a beta-subunit of an acetyl-CoA carboxylase (acetyl-CoA carboxylase, beta subunit) having a nucleotide sequence corresponding to that shown in SEQ ID NO. At least 74% identity, and more preferably at least 75%, 80%, 85%, 87.5%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity;
  • a beta subunit of an acetyl-CoA carboxylase (acetyl-CoA carboxylase, beta subunit) AccD having an amino acid sequence corresponding to that shown in SEQ ID NO. 264 has at least 94% identity and more preferably at least 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid acpA encoding an acyl carrier protein having a nucleotide sequence corresponding to that shown in SEQ ID NO. At least 93% identity, and more preferably at least 94%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • an acyl carrier protein AcpA having an amino acid sequence corresponding to the amino acid sequence shown in SEQ ID NO. 266 has at least 94% identity and more preferably at least 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity;
  • a nucleic acid acpS coding for a holo-acyl carrier protein synthase having a nucleotide sequence which corresponds to that shown in SEQ ID NO. At least 32% identity and more preferably at least 40%, 50%, 60%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% and most preferably 100 % Identity;
  • holo-acyl carrier protein synthase (holo-acyl carrier protein synthase)
  • AcpS having an amino acid sequence corresponding to the amino acid sequence shown in SEQ ID NO. 268 has at least 94% identity and more preferably at least 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid birA coding for a transcriptional repressor of the biotin operon (transcriptional repressor of the biotin operon), having a nucleotide sequence which corresponds to that shown in SEQ ID NO. 269 nucleotide sequence has at least 72% identity, and more preferably at least 75%, 80%, 85%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity
  • a transcriptional repressor of the biotin operon (transcriptional repressor of the biotin operon) BirA having an amino acid sequence corresponding to that shown in SEQ ID NO. 270 has at least 94% identity and more preferably at least 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • a nucleic acid fabD coding for a malonyl CoA-acyl carrier protein transacylase having a nucleotide sequence which corresponds to that shown in SEQ ID NO. At least 78% identity and more preferably at least 80%, 85%, 87.5%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity are given having;
  • a malonyl CoA-acyl carrier protein transacylase (malonyl CoA acyl carrier protein transacylase) FabD having an amino acid sequence corresponding to that shown in SEQ ID NO. 272 has at least 94% identity and more preferably at least 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid fabF encoding a 3-oxoacyl (acyl carrier protein) synthase (3-oxoacyl (acyl-carrier protein) synthase) having a nucleotide sequence corresponding to that shown in SEQ ID NO. At least 83% identity, and more preferably at least 85%, 87.5%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • a 3-oxoacyl (acyl carrier protein) synthase (3-oxoacyl (acyl carrier protein) synthase) FabF having an amino acid sequence identical to that shown in SEQ ID NO. 274 has at least 94% identity and, more preferably, at least 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity;
  • nucleic acid fabG encoding a beta-ketoacyl-acyl carrier protein reductase (beta-ketoacyl-acyl carrier protein reductase) having a nucleotide sequence corresponding to that shown in SEQ ID NO. At least 77% identity and more preferably at least 80%, 85%, 87.5%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity having;
  • a beta-ketoacyl-acyl carrier protein reductase (beta-ketoacyl-acyl carrier protein reductase) FabG having an amino acid sequence which corresponds to the amino acid sequence shown in SEQ ID NO. 276 has at least 94% identity and more preferably at least 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • a nucleic acid cdsA encoding a phosphatidate cytidylyl transferase (phosphatidate cytidylyltransferase; EC 2.7.7.41) having a nucleotide sequence corresponding to that shown in SEQ ID NO. 277 has at least 69% identity and more preferably at least 70%, 80%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • CdsA having an amino acid sequence corresponding to that shown in SEQ ID NO. 278 has at least 94% identity and more preferably at least 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • NAD H-dependent glyceryl-3-phosphate dehydrogenase
  • NAD H-dependent glycerol-3-phosphate dehydrogenase
  • GpsA having an amino acid sequence identical to that shown in SEQ ID NO. 280 has at least 81% identity, and more preferably at least 85%, 90%, 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity;
  • nucleic acid pgsA encoding a phosphatidyl-glycerophosphate synthase (phosphatidylglycerophosphate synthase) having a nucleotide sequence corresponding to that shown in SEQ ID NO. At least 79% identity, and more preferably at least 80%, 85%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • phosphatidyl-glycerophosphate synthase phosphatidylglycerophosphate synthase
  • PgsA having an amino acid sequence identical to that shown in SEQ ID NO. At least 88% identity, and more preferably at least 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • an i-acylglyceryl-3-phosphate (i-acylglycerol-3-phosphate) YhdO having an amino acid sequence corresponding to that shown in SEQ ID NO. 284 has at least 94% identity and, more preferably, at least 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity;
  • nucleic acid pIsX coding for a protein involved in fatty acid / phospholipid synthesis having a nucleotide sequence which corresponds to that shown in SEQ ID NO. At least 77% identity, and more preferably at least 80%, 85%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • a protein involved in fatty acid / phospholipid synthesis PIsX having an amino acid sequence corresponding to that shown in SEQ ID NO. 286 has at least 94% identity and, more preferably, at least 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity;
  • a nucleic acid gcaD encoding a UDP-N-acetyl-glucosamine pyrophosphorylase (UDP-N-acetylglucosamine pyrophosphorylase) having a nucleotide sequence corresponding to that shown in SEQ ID NO. At least 77% identity and more preferably at least 80%, 82.5%, 85%, 87.5%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and 287 nucleotide sequence most preferably 100% identity;
  • UDP-N-acetyl-glucosamine pyrophosphorylase UDP-N-acetyl-glucosamine pyrophosphorylase
  • GcaD having an amino acid sequence corresponding to that shown in SEQ ID NO. 288 has at least 94% identity and, more preferably, at least 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity;
  • a glmS nucleic acid encoding a glucosamine-fructose-6-phosphate-amino-transferase (glucosamine-fructose-6-phosphate aminotransferase) having a nucleotide sequence corresponding to that shown in SEQ ID NO. At least 84% identity, and more preferably at least 85%, 87.5%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • a glucosamine-fructose-e-phosphate-amino-transferase glucosamine-fructose-6-phosphate aminotransferase
  • GImS having an amino acid sequence corresponding to that shown in SEQ ID NO. 290 has at least 94% identity and more preferably at least 95%, 96%, 97%, 98%, 99% and most preferably 100% identity
  • a nucleic acid ybbT coding for a phospho-glucosamine mutase phosphoglucosamine mutase; EC 5.4.2.10 having a nucleotide sequence which corresponds to that shown in SEQ ID NO. At least 82% identity and more preferably at least 85%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • a phospho-glucosamine mutase (phosphoglucosamine mutase; E.C. 5.4.2.10) YbbT having an amino acid sequence identical to that shown in SEQ ID NO. 292 has at least 94% identity and, more preferably, at least 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity;
  • a yvyH nucleic acid encoding a putative UDP-N-acetylglucosamine 2-epimerase (putative UDP-N-acetylglucosamine 2-epimerase; E.C. 5.1.3.14) having a nucleotide sequence corresponding to that shown in SEQ ID NO. At least 79% identity and more preferably at least 80%, 85%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • YvyH having an amino acid sequence corresponding to that shown in SEQ ID NO. 294 has at least 89% identity and more preferably at least 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • nucleic acid asd coding for an aspartate semialdehyde dehydrogenase having a nucleotide sequence which corresponds to that shown in SEQ ID NO. At least 77% identity, and more preferably at least 80%, 85%, 87.5%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity having;
  • an aspartate semialdehyde dehydrogenase Asd having an amino acid sequence identical to that shown in SEQ ID NO. 296 has at least 89% identity, and more preferably at least 90%, 92.5%, 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity;
  • a nucleic acid dapA encoding a dihydrodipicolinate synthase (dihydrodipicolinate synthase) having a nucleotide sequence corresponding to that shown in SEQ ID NO. At least 77% identity, and more preferably at least 80%, 85%, 87.5%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity having;
  • a dihydrodipicolinate synthase (dihydrodipicolinate synthase) DapA having an amino acid sequence identical to that shown in SEQ ID NO. 298 has at least 84% identity and more preferably at least 85%, 87.5%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99%, and most preferably 100% identity;
  • a dapB nucleic acid encoding a dihydrodipicolinate reductase (dihydrodipicolinate reductase) having a nucleotide sequence corresponding to that shown in SEQ ID NO. At least 80% identity, and more preferably at least 85%, 90%, 92.5%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity;
  • dihydrodipicolinate reductase (dihydrodipicolinate reductase) DapB having an amino acid sequence identical to that shown in SEQ ID NO. Has at least 88% identity and more preferably at least 90%, 92%, 94%, 95%, 96%, 97%, 98%, 99% and most preferably 100% identity.
  • nucleic acids given in the sequence listing may also be used as DNA probes to detect the respective homologous genes in preparations of genomic DNA of other species.
  • the procedure for this is also known per se; as well as the isolation of the genes thus obtained, their cloning, their expression and recovery of the associated proteins.
  • it is intended to work such as those shown in Example 2 for B. licheniformis itself.
  • nucleic acids defined with reference to SEQ ID NO. 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149, 151, 153, 155, 157, 159, 161, 163, 165, 167, 169, 171, 173, 175, 177, 179, 181, 183, 185, 187
  • the gene products and / or nucleic acids from natural Species in particular to obtain microorganisms.
  • those which are capable of fermentation and which are actually used in large-scale fermentations are increasingly preferred with regard to the stated task.
  • These include in particular representatives of the genera Staphylococcus, Corynebacterium and Bacillus. These include, for example, S. carnosus and C. glutamicum, as well as B. subtilis, B. licheniformis, B. amyloliquefaciens, B. agaradherens, B. lentus, B. globigii and ⁇ . alkalophilus.
  • B. licheniformis DSM13 because from it the exact sequences listed in the Sequence Listing could be obtained.
  • nucleic acids coding for a gene product in each case those which are described for one of the previously described, with reference to SEQ ID NO. 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150, 152, 154, 156, 158, 160, 162, 164, 166, 168, 170, 172, 174, 176, 178, 180, 182,
  • the Proteins of the invention are also available for their positive use. In this respect, it is important to be able to produce these biotechnologically by methods known per se.
  • the respective associated nucleic acids are used for this purpose.
  • a further subject of the invention represent vectors which have one of the previously described, with reference to SEQ ID NO. 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149, 151, 153, 155, 157, 159, 161, 163, 165, 167, 169, 171, 173, 175, 177, 179, 181, 183, 185, 187, 189, 19
  • vectors are commercially available in large numbers and with a wide variety of variations, both for cloning and for expression. These include, for example, vectors derived from bacterial plasmids, bacteriophages or viruses, or predominantly synthetic vectors. Further, they will be after the type of cell types in which they establish themselves For example, a distinction is made between vectors for Gram-negative, for Gram-positive bacteria, for yeast or for higher eukaryotes. They form suitable starting points for example for molecular biological and biochemical investigations and for the expression of the relevant gene or associated protein. In particular, for the production of constructs for the deletion or enhancement of expression, they are practically indispensable, as is apparent from the relevant prior art.
  • the relevant genes are storable simultaneously or can be expressed under the control of the same promoter.
  • a vector containing simultaneously two or more intact copies of the genes of the invention can serve to rescue a deletion mutant which is simultaneously deleted in several of these genes (rescue). Targeted removal of this vector will then result in the simultaneous shutdown of these multiple genes, resulting in lethality in the case of essential genes.
  • vectors of the invention are cloning vectors.
  • cloning vectors are suitable in addition to the storage, the biological amplification or the selection of the gene of interest for its molecular biological characterization. At the same time, they are transportable and storable forms of the claimed nucleic acids and are also starting points for molecular biological techniques that are not bound to cells, such as PCR or in vitro mutagenesis methods.
  • vectors according to the invention are expression vectors.
  • expression vectors are the basis for realizing the corresponding nucleic acids in biological production systems and thus to produce the associated proteins, since they allow in vivo transcription and translation, that is, the synthesis of the relevant gene product.
  • expression vectors corresponding to the Expression carry necessary genetic elements, for example, the natural, originally located in front of this gene promoter or a promoter from another organism. These elements can be arranged for example in the form of a so-called expression cassette. Alternatively, individual or all regulatory elements can also be provided by the respective host cell. With particular preference, the expression vectors are matched to the selected expression system, in particular the host cell (see below), with regard to further properties, for example the optimal copy number.
  • a separate subject of the invention form cells which, after .Gentechnischer modification one of the previously designated, with reference to SEQ ID NO. 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149, 151, 153, 155, 157, 159, 161, 163, 165, 167, 169, 171, 173, 175, 177, 179, 181, 183, 185,
  • these cells contain the genetic information for the synthesis of a protein according to the invention.
  • suitable suitable host cells are those which can be cultivated relatively easily and / or yield high product yields. They allow, for example, the amplification of the corresponding genes, but also their mutagenesis or transcription and translation and ultimately the biotechnological production of the proteins in question.
  • This genetic information can either be extrachromosomally as a separate genetic element, ie be present in bacteria in plasmidaler localization or integrated into a chromosome.
  • the choice of a suitable system depends on issues such as the nature and duration of storage of the gene, or the organism or the type of mutagenesis or selection. Such implementation possibilities are known per se to the molecular biologist.
  • nucleic acid is part of a vector, in particular a vector according to the invention described above.
  • Preferred among these cells is in each case a host cell which is a bacterium.
  • bacteria are characterized by short generation times and low demands on the cultivation conditions. As a result, inexpensive methods can be established. In addition, bacteria have a wealth of experience in fermentation technology. For a specific production gram-negative or gram-positive bacteria may be suitable for a variety of reasons to be determined experimentally in individual cases, such as nutrient sources, product formation rate, time requirement, etc.
  • it is a gram-negative bacterium, in particular one of the genera Escherichia coli, Klebsiella, Pseudomonas or Xanthomonas, in particular strains of E. coli K12, E. coli B or Klebsiella planticola, and more particularly derivatives of the strains Escherichia coli BL21 (DE3), E. coli RV308, E. coli DH5 ⁇ , E. coli JM109, E. coli XI ⁇ or Klebsiella planticola (Rf).
  • Gram-negative bacteria such as E. coli, a variety of proteins are secreted into the periplasmic space. This can be advantageous for special applications.
  • the application WO 01/81597 A1 discloses a method according to which it is achieved that gram-negative bacteria also eject the expressed proteins.
  • the Gram-negative bacteria which are mentioned as preferred are generally light, that is to say commercially or accessible via public strain collections and, in conjunction with other components which are likewise available in large numbers, such as vectors, can be optimized for specific production conditions.
  • it is a Gram-positive bacterium, in particular one of the genera Bacillus, Staphylococcus or Corynebacterium, more particularly of the species Bacillus lentus, B. licheniformis, B. amyloliquefaciens, B.subtilis, B. globigii or B. alcalophilus, Staphylococcus carnosus or Corynebacterium glutamicum, and among these most preferably a derivative of B.licheniformis DSM 13.
  • Gram-positive bacteria have the Gram-negative compared to the fundamental difference to release secreted proteins immediately into the nutrient medium surrounding the cells, from which, if desired, can directly purify the expressed proteins of the invention.
  • they are related or identical to most of the organisms of origin for technically important enzymes and usually form even comparable enzymes, so they have a similar codon Usage and their protein synthesizer is naturally aligned accordingly.
  • Very particular preference is given to derivatives of B. licheniformis DSM 13 because they are also widely used in the art as biotechnological production strains and because, on the other hand, with the provisional applications exactly the genes and proteins according to the invention from B.licheniformis DSM 13 are available so that the realization of the present invention should be most successful in such strains.
  • Another embodiment of the present invention provides methods of making one or more of those described above with reference to SEQ ID NO. 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150, 152, 154, 156, 158, 160, 162, 164, 166, 168, 170, 172, 174, 176, 178, 180, 182, 184, 186, 188, 190, 192, 194,
  • an enzymatic detection of the enzyme activities concerned by suitable detection reactions is done, for example, so that the relevant for the reaction in question starting compound presented, incubated with a sample of the supernatant (containing the secreted enzymes) or a cell extract (if they are not secreted) and the product formed is detected by any appropriate measurement methods.
  • proteins shown in the present sequence listing can be synthesized by conventional methods and antibody can be generated against this; this also applies to proteins that are not recognizable by a specific enzymatic reaction. These proteins are then detectable, for example via corresponding Western blots.
  • the correspondingly preferred genetic information is provided in microbiologically utilizable form, that is to say for genetic engineering production methods.
  • Increasingly preferred is the provision of a particularly successfully utilizable by the host cell vector or of such cells themselves.
  • the relevant production methods are known in the art per se.
  • Embodiments of the present invention may also be cell-free expression systems in which protein biosynthesis is understood in vitro. All of the elements already described above can also be combined to form new methods for producing proteins according to the invention. It is conceivable for each protein according to the invention a variety of possible combinations of process steps, so that optimal procedures must be determined experimentally for each specific case.
  • nucleotide sequence has been adapted in one, preferably more and more preferably all codons to the codon Usage of the host strain.
  • a further subject of the invention is the use of an above with reference to SEQ ID NO. 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150, 152, 154, 156, 158, 160, 162, 164, 166, 168, 170, 172, 174, 176, 178, 180, 182, 184, 186, 188, 190, 192, 194, 196, 198
  • an example of such uses is the overexpression of factors of protein biosynthesis in cells used as expression systems.
  • the protein biosynthetic performance of these cells can be increased by overexpressing one or, preferably, several ribosomal proteins; those are above with reference to SEQ ID NO. 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132 and 134 (large subunit) or with reference to SEQ ID NO.
  • an essentially equivalent, preferably combinable effect should result from the overexpression of aminoacyl tRNA synthetases, as described above with reference to SEQ ID NO. 176, 178, 180, 182, 184, 186, 188, 190, 192, 194, 196, 198, 200, 202, 204, 206, 208, 210, 212, 214, 216, 218 or 220, and / or translation factors as described above with reference to SEQ ID NO.
  • alpha chain of the DNA-dependent RNA polymerase (SEQ ID NO: 48), beta chain of the DNA-dependent RNA polymerase (SEQ ID NO: 50), beta DNA-dependent RNA polymerase chain (SEQ ID NO: 52), Greater RNA polymerase sigma factor 43 (SEQ ID NO: 54) and / or the polyA polymerase (SEQ ID NO: 56).
  • a preferred such use is one which is an in vitro approach.
  • DNA preparations using a DNA helicase defined above with reference to SEQ ID NO. 6, a DNA ligase, defined above with reference to SEQ ID NO. 22, a DNA single-strand binding protein, defined above with reference to SEQ ID NO. 30, or a DNA topoisomerase, defined above with reference to SEQ ID NO. 44;
  • amino acid translation systems enriched for aminoacyl tRNA synthetases as described above with reference to SEQ ID NO. 176, 178, 180, 182, 184, 186, 188, 190, 192, 194, 196, 198, 200, 202, 204, 206, 208, 210, 212, 214, 216, 218 or 220; - / n-translation systems further enriched for translation factors as described above with reference to SEQ ID NO. 226, 228, 230, 232, 234, 236 or 238 are defined;
  • Another embodiment of the present invention is the use of a reference to SEQ ID NO. 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149, 151, 153, 155, 157, 159, 161, 163, 165, 167, 169, 171, 173, 175, 177, 179, 181, 183, 185, 187, 189, 191, 193, 195,
  • functional inactivation means any type of modification or mutation which prevents the function of the relevant protein from being inhibited.
  • a particular "use" of these factors or genes of this embodiment resides in the fact that they no longer naturally function in the cell in question.This is achieved according to this subject matter at the genetic level by shutting off the gene in question , This procedure requires a special safety precaution, because in all cases and according to the invention it is an essential gene whose absence is directly lethal for the cell in question.
  • an intact copy of the same gene must be introduced into the cell in question, preferably one own genetic element, that is a plasmid.
  • the essential systemic advantage of this selection method against selection via resistance genes to toxins, that is to say antibiotics, is that it is not necessary to permanently add a corresponding substance during the cultivation and that reversion to the wild type is less frequent. This is associated with the other, initially described advantages.
  • the essential systemic advantage of this selection method over a selection by auxotrophy is that it is not necessary to dispense with complex, that is low-priced, high-availability nutrient media during cultivation, because the respective deficiency can not be compensated by the intake of a simple nutrient. This is particularly clear when one realizes that, for example, with the (even-numbered) SEQ ID NO. 74 to 174 (and the respective homologous gene products) ribosomal proteins are referred to, without which in no case functional ribosomes can be assembled, which perform the protein biosynthesis.
  • the functional inactivation results in that the protein in question is not synthesized in full length, preferably not at all.
  • these uses are those in which the functional inactivation occurs during the fermentation of the microorganism. Because of the task, the fermentation of the microorganisms used for biotechnological production should be improved.
  • the activities or protein contents clearly attributable to the relevant, inactivated enzymes or proteins are reduced to less than 50%, preferably to less than 20%, very particularly preferably to less than 5% of the activity or concentration values occurring without the inactivation reduced.
  • This grading with regard to the degree of inactivation takes into account the different effectiveness of the various methods available for inactivation.
  • cells of a non-treated strain and a treated strain are fermented under otherwise identical conditions, and suitably, during fermentation, the enzyme activities concerned are determined suitably from cell extracts. Since the strains are otherwise identical, the differences in activity are attributable to the inactivations according to the invention. In this case, according to the invention, any corresponding reduction in activity is desired. Percentage comparable values are obtained by taking samples from both fermentations and determining the activities in question by methods known per se.
  • the respective value determinable in the sample according to the invention is less than 50%, 40%, 30%, 20%, 10%, 7.5, 5% and especially less than 1% when moving to the stationary growth phase. of the corresponding value of the reference bond.
  • an antibody-based detection reaction can be carried out analogously, as has already been described in principle above.
  • 2, 3 or more of said genes are functionally inactivated.
  • this serves to actually produce a lethal genotype.
  • RNA interference For some approaches, such as RNA interference, one can not always assume a 100% success.
  • this serves to establish several plasmids in the same cell simultaneously according to the principle described above. This is especially dissuaded when several transgenes are to be introduced next to each other, the DNA segments of interest do not have space next to each other on the same plasmid or should come into effect separately from each other.
  • the selection pressure is that each plasmid will cure one of the said genetic defects.
  • these functional inactivation uses are such uses, wherein for the inactivation each one used for a non-active protein nucleic acid with a point mutation.
  • nucleic acids can be generated by per se known methods for point mutagenesis. Such are, for example, in relevant handbooks such as those by Fritsch, Sambrook and Maniatis, "Molecular cloning: a laboratory manual", CoId Spring Harbor Laboratory Press, New York, 1989. In addition, there are now numerous commercial kits available, such as the QuickChange . ® kit from Stratagene, La JoIIa, USA Its principle is that, oligonucleotides are synthesized with each exchange (mismatch primer) and hybridized with the gene in single;.
  • a nucleic acid with a deletion or insertion mutation is used in each case for the functional inactivation, preferably comprising the border sequences of the region coding for the protein comprising at least 70 to 150 nucleic acid positions.
  • a match is in each case at least 70 to 150 contiguous nucleic acid positions, in each case in the two border sequences to the non-coincident one Part necessary, where it does not depend on the intermediate part. Accordingly, those embodiments are preferred which comprise only two flanking regions of at least these sizes.
  • nucleic acids having a total of two nucleic acid sections are used, which each comprise at least 70 to 150 nucleic acid positions and thus at least partially, preferably completely, flank the region coding for the protein.
  • the flanking regions can be determined starting from the known sequences by methods known per se, for example by means of outwardly directed PCR primers and a preparation of genomic DNA as a template (anchored PCR). Because only to allow the exchange of the two gene copies via homologous recombination, it does not necessarily need to be protein coding sections.
  • the primers required for this purpose can be designed on the basis of the nucleotide sequences given in the sequence listing also for other species of Gram-positive bacteria and hereof in particular for those of the genus Bacillus.
  • such at least partially non-coding regions for many of these genes can be obtained from related species, for example from B. subtilis database entries, for example the Subtilist database of the Institut Pasteur, Paris, France (http: //genolist.pasteur. fr / Subtilist / genome.cgi).
  • a nucleic acid which is identical or interferes with the 5'-terminal sequence, in particular the signal sequence of the associated gene or parts thereof, is used for the functional inactivation.
  • One embodiment of the functional inactivation uses described heretofore are those uses where, for curing the defect, a nucleic acid coding for an active protein is introduced on a vector into the host cell, preferably on a vector posing as a stable genetic element , particularly preferably as a plasmid in the cell established.
  • this is such a use, wherein the nucleic acid responsible for said curing contains, in addition to the protein-only coding portion, the naturally-linked promoter or a promoter-sufficient portion thereof.
  • Activation of a gene means that it produces enough protein under the appropriate conditions.
  • constitutive promoters that is, those which are virtually permanently active.
  • An optimal curing of the gene defect induced according to the invention should be equivalent in particular to the naturally prevailing situation if the gene in question is regulated in much the same way as in natural, chromosomal localization.
  • a further embodiment of the invention intended for this purpose is such a use, wherein on the same genetic element as that for the said curing of the defect a genetic information to be selected lies, preferably under the control of the same promoter.
  • a genetic information of particular interest to be selected is stably established in a cell line by being introduced on such a curing vector into a cell having the otherwise lethal gene defect, which is only cured by the presence of this vector.
  • both genetic elements are under the control of the same promoter, the effect is exploited that it is in particular in such constitutive promoters to virtually permanently active and often strong promoters.
  • Another advantage may be that no additional promoter needs to be provided for the transgene; As a result, the construction of the relevant vector is comparatively simple.
  • Preferred embodiments of the uses described herein are such uses, wherein the genetic information to be selected is that for a protein, for an enzyme and / or for information for the synthesis of a low molecular weight compound.
  • the present invention is also realized in the form of genetically modified microorganisms, to which the above applies accordingly.
  • the microorganisms in which the chromosomal copy is at least one of the above with reference to SEQ ID NO. 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149, 151, 153, 155, 157, 159, 161, 163, 165, 167, 169, 171, 173, 175, 177, 179, 181, 183, 185,
  • strains can be obtained in this way in which several otherwise lethal gene defects are cured via several simultaneously existing plasmids.
  • Each of these plasmids can carry transgenes of experimental or technical interest.
  • Microorganisms according to the invention are preferably those in which a nucleic acid coding for an active protein has been introduced into the host cell on a vector for curing the defect, preferably on a vector which is a stable genetic element, particularly preferably a plasmid in the cell has established. For this results in the described effect, according to which the genetic information necessary for curing remains stable over generations in the relevant cell lines.
  • microorganisms according to the invention are preferably those in which the nucleic acid responsible for said curing contains, in addition to the protein-coding portion, the naturally associated promoter or a promoter portion thereof.
  • microorganisms according to the invention are those in which a genetic information to be selected is located on the same genetic element as that for said curing of the defect, preferably under the control of the same promoter.
  • microorganisms according to the invention are preferably those where the genetic information to be selected is that for a protein, for an enzyme and / or for the synthesis of a low molecular weight compound.
  • Microorganisms according to the invention are preferably those which are bacteria.
  • those microorganisms are preferred which are gram-negative bacteria, in particular those of the genera Escherichia coli, Klebsiella, Pseudomonas or Xanthomonas, in particular strains of E. coli K12, E. coli B or Klebsiella planticola, and all especially derivatives of strains Escherichia coli BL21 (DE3), E. coli RV308, E. coli DH5 ⁇ , E. coli JM109, E. coli XL-1 or Klebsiella planticola (Rf).
  • those microorganisms which are Gram-positive bacteria are no less preferred, in particular those of the genera Bacillus, Staphylococcus or Corynebacterium, more particularly of the species Bacillus lentus, B. licheniformis, B. amyloliquefaciens, B. subtilis, B Globigii or B. alcalophilus, Staphylococcus carnosus or Corynebacterium glutamicum, and most particularly B. licheniformis DSM 13.
  • the invention is realized in particular in corresponding fermentation process according to the invention.
  • the method is wherein the low molecular weight compound is a natural product, a nutritional supplement, or a pharmaceutically-relevant compound.
  • amino acids, vitamins or oligopeptides are produced which find particular use as food supplements.
  • Pharmaceutically relevant compounds may be precursors or intermediates Medications or even act on these themselves. In all these cases one also speaks of biotransformation, according to which the metabolic properties of the microorganisms are exploited in order to replace the otherwise complex chemical synthesis completely or at least in individual steps.
  • the corresponding method is where the protein thus formed is a peptide hormone.
  • peptide hormones such as interleukins or insulin are obtained on a large scale by fermentations, often by eukaryotic host cells.
  • the protein formed in this way is an enzyme, in particular one from the group of ⁇ -amylases, proteases, cellulases, lipases, oxidoreductases, peroxidases, laccases, oxidases and hemicellulases.
  • Industrial enzymes produced by such processes find use, for example, in the food industry.
  • ⁇ -amylases are used to prevent the staling of bread or to clarify fruit juices.
  • Proteases are used to digest proteins. All of these enzymes are described for use in detergents and cleaning agents, in particular the subtilisin proteases already produced naturally by Gram-positive bacteria occupy a prominent place. Especially in the textile and leather industries, they serve the processing of natural resources. Furthermore, all these enzymes can be used in turn in the sense of biotransformation as catalysts for chemical reactions.
  • Another embodiment of the present invention is the use of a reference to SEQ ID NO. 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149, 151, 153, 155, 157, 159, 161, 163, 165, 167, 169, 171, 173, 175, 177, 179, 181, 183, 185, 187, 189, 191, 193, 195,
  • promoters associated with the nucleic acids according to the invention are not given in the sequence listing, but can easily be obtained by methods known per se, in particular by making use of the protein-coding sequences disclosed therein. These sequences can be used to obtain the associated promoters, in particular from B. licheniformis DSM12 and, in principle, all species related thereto, the prospects of success, as described above for the protein-coding regions, being the higher, the closer the organisms in question are to B.licheniformis.
  • promoters obtained thereby are also available for the preparation of further constructs, in particular of expression vectors. These include, in particular, those with which gene products according to the invention can be prepared in large quantities as described above, for example, in order to allow them to be used in vitro.
  • said nucleic acid is hybridized with a preparation of genomic DNA.
  • This genomic DNA is derived, for example, from the host to which the above-described inactivation according to the invention is to be carried out. It is prepared in a manner known per se and investigated in a Southem hybridization experiment with the nucleic acid according to the invention used as a probe. From the fragment thus identified, the 5'-located portion can be isolated by also known methods, advantageously in the context of the associated gene, so that the promoter gene fragment thus obtained can be introduced into a corresponding caging vector.
  • An alternative use for this purpose is to form primers based on said nucleic acid, via which flanking DNA sections from a preparation of a genomic DNA are identified.
  • DSM 13 German Collection of Microorganisms and Cell Cultures GmbH (DSMZ) 1 Mascheroder Weg 1 b, 38124 Braunschweig (http://www.dsmz.de).
  • S. licheniformis is dissolved in a 500 ml shake flask in 100 ml Horikoshi medium pH 9 (0.1% K 2 HPO 4 , 0.5% yeast extract, 1% peptone, 0.02% MgSO 4 , 0.3% Na 2 CO 3 ) for 72 h at 37 ° C and 200 rpm cultivated.
  • the cells are separated from the supernatant by centrifugation. The supernatant contains the secreted proteins.
  • the genomic DNA was prepared by standard methods, mechanically fractionated and separated by electrophoresis in a 0.8% agarose gel.
  • the 2 to 2.5 kb fragments were eluted from the agarose gel, dephosphorylated and ligated as blunt ended fragments into the Smal restriction site of the vector pTZ19R-Cm.
  • This is a chloramphenicol-resistance-conferring derivative of the commercially available from the company Fermentas (St. Leon-Red) plasmid pTZ19R. This gave a library of the smaller fragments.
  • the genomic fragments obtained by partial restriction with the enzyme Sau 3A1 were ligated into the Supercos-1 vector system ("cosmid vector kit") of the company Stratagene, La JoIIa, USA, whereby a gene bank of the predominantly larger fragments was obtained.
  • the bacteria E. coli DH5 ⁇ (D. Hannahan (1983): “Studies on Transformation on Escherichia coli", J. Mol. Microbiol., Vol. 166, pp. 557-580) which were obtainable by transformation with the relevant gene banks, were used for the respective recombinant plasmids isolated and sequenced.
  • the dye terminator chemistry was used, performed by the automatic sequencers Mega-BACE 1000/4000 (Amersham Bioscence, Piscataway, USA) and ABI Prism 377 (Applied Biosystems, Foster City, USA).
  • sequences indicated in the sequence listing of the present application SEQ ID NO. 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149, 151, 153, 155, 157, 159, 161, 163, 165, 167, 169, 171, 173, 175, 177, 179, 181, 183, 185, 187, 189, 191, 191, 19
  • GenBank National Center for Biotechnology Information, NCBI, National Institutes of Health, Bethesda, MD, USA, http://www.ncbi.nlm.nih.gov was searched by the databases. and subtilist of the Institute Pasteur, Paris, France
  • Table 1 Near-like genes or proteins to the genes and proteins identified in Example 2.

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Abstract

La présente invention concerne 150 nouveaux gènes essentiels et leurs produits génétiques de Bacillus licheniformis, et des gènes et protéines analogues appropriés. Ces facteurs prennent en charge des processus vitaux respectivement élémentaires in vivo, par exemple la réplication (par ex. ADN-polymérase, hélicase ou gyrase), la transcription (par ex. ARN-polymérase), la biosynthèse protéique (protéines ribosomales, aminocyl-ARNt-synthétases, facteurs d'initiation et d'élongation), la sécrétion de protéines (par ex. translocase) ou le métabolisme énergétique. Un manquement de ces gènes est directement létal pour les cellules concernées, et ne peut pas être compensé par d'éventuels composés provenant du milieu nutritif. Les gènes associés peuvent ainsi être utilisés comme marqueurs de sélection. Il est possible d'établir des procédés de production biotechnologiques améliorés par des micro-organismes, dans la mesure où une sélection par l'intermédiaire de ces gènes peut s'effectuer sans antibiotique, et les composés de milieux nutritifs courants ne permettent pas de compensation.
PCT/EP2005/008683 2004-08-19 2005-08-10 Nouveaux genes essentiels de bacillus licheniformis et procede de production biotechnologiques ameliores, bases sur ceux-ci Ceased WO2006018205A2 (fr)

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2006112738A1 (fr) * 2005-04-19 2006-10-26 Auckland Uniservices Limited Nouveaux peptides et méthodes pour le traitement de troubles inflammatoires
WO2008066931A3 (fr) * 2006-11-29 2008-07-31 Novozymes Inc Chromosome de bacillus licheniformis
JP2023534879A (ja) * 2020-07-24 2023-08-14 上海交通大学 Ｎ－アセチルグルコサミン産生菌株並びにその構築方法及び使用

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ES2188190T5 (es) 1998-07-21 2007-11-16 Danisco A/S Producto alimentario.
DK1387616T3 (da) 2001-05-18 2007-09-24 Danisco Fremgangsmåde til fremstilling af en dej med et enzym
US20050196766A1 (en) 2003-12-24 2005-09-08 Soe Jorn B. Proteins
US7955814B2 (en) 2003-01-17 2011-06-07 Danisco A/S Method
MX2009008021A (es) 2007-01-25 2009-08-07 Danisco Produccion de una aciltransferasa de lipido de celulas de bacillus licheniformis transformadas.

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ES2079348T3 (es) * 1986-08-05 1996-01-16 Transgene Sa Procedimiento de estabilizacion de un plasmido contenido en una cepa bacteriana y cepa obtenida.
EP0972838B1 (fr) * 1998-07-15 2004-09-15 Roche Diagnostics GmbH Système de sélection par complémentation d' une auxotrophie en absence d'antibiotique chez E. coli
DE10309557A1 (de) * 2003-03-04 2004-09-23 Henkel Kgaa Ein Translokationsenzym als Selektionsmarker

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2006112738A1 (fr) * 2005-04-19 2006-10-26 Auckland Uniservices Limited Nouveaux peptides et méthodes pour le traitement de troubles inflammatoires
WO2008066931A3 (fr) * 2006-11-29 2008-07-31 Novozymes Inc Chromosome de bacillus licheniformis
JP2023534879A (ja) * 2020-07-24 2023-08-14 上海交通大学 Ｎ－アセチルグルコサミン産生菌株並びにその構築方法及び使用
JP7742399B2 (ja) 2020-07-24 2025-09-19 上海交通大学 Ｎ－アセチルグルコサミン産生菌株並びにその構築方法及び使用

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