CA2322719A1 - Genetic sequences conferring pathogen resistance in plants and uses therefor - Google Patents

Abstract

The present invention provides isolated genetic sequences derived from barley and maize that confer, or otherwise facilitate or enhance, resistance in plants to plant pathogens, in particular, against Puccinia sorghi and barley stem rust, and genetic constructs comprising said genetic sequences. The present invention further provides for plants into which the subject genetic sequences have been introduced, generating enhanced resistance qualities to plant fungal pathogens and for methods of producing same.

Description

GENETIC SEQUENCES CONFERRING PATHOGEN RESISTANCE
IN PLANTS AND USES THEREFOR
FIELD OF THE INVENTION
The present invention relates generally to isolated genetic sequences derived from plants and more particularly to isolated genetic sequences derived from plants which confer, or otherwise facilitate or enhance, resistance in plants to plant fungal pathogens, such as fungi, rusts, nematodes, viruses and bacterial pathogens.
The present invention further provides for plants into which the subject genetic sequences have been introduced, generating enhanced resistance qualities to plant fungal pathogens. The present invention is particularly useful in the development of plants which have improved resistance to plant fungal pathogens, in particular cereal crop plants which have improved resistance to rusts.
GENERAL
Bibliographic details of the publications referred to in this spec~cation are collected at the end of the description.
As used herein the term "derived from" shall be taken to indicate that a specified integer may be obtained from a particular specified source or species, albeit not necessarily directly from that specified source or species.
Throughout this specifiication, unless the context requires otherwise, the word "comprise", or variations such as "comprises" or "comprising", will be understood to imply the inclusion of a stated step or element or integer or group of steps or elements or integers but not the exclusion of any other step or element or integer or group of elements or integers.
Those skilled in the art will appreciate that the invention described herein is susceptible to variations and modifications other than those specifically described. It is to be understood that the invention includes all such variations and modifications. The invention also includes all of the steps, features, compositions and compounds referred to or indicated in this specification, individually or collectively, and any and all combinations or any two or more of said steps or WO 99/4511$ PCT/AU99100130 features.
The present invention is not to be limited in scope by the specific embodiments described herein, which are intended for the purposes of exemplification only.
Functionally-equivalent products, compositions and methods are clearly within the scope of the invention, as described herein.
Sequence identity numbers (SEQ 1D NOS.) containing nucleotide and amino acid sequence information included in this specification are collected after the Abstract and have been prepared using the programme Patentln Version 2Ø Each nucleotide or amino acid sequence is identified in the sequence listing by the numeric indicator <210> followed by the sequence identifier (e.g. <210>1, <210>2, etc). The length, type of sequence (DNA, protein (PRT), etc) and source organism for each nucleotide or amino acid sequence are indicated by information provided in the numeric indicator fields <211 >, <212> and <213>, respectively. Nucleotide and amino acid sequences referred to in the specification are defined by the information provided in numeric indicator field <400> followed by the sequence identifier (eg. <400>1, <400>2, etc).
The designation of nucleotide residues referred to herein are those recommended by the IUPAC-lUB Biochemical Nomenclature Commission, wherein A represents Adenine, C
represents Cytosine, G represents Guanine, T represents thymine, Y represents a pyrimidine residue, R represents a purine residue, M represents Adenine or Cytosine, K
represents Guanine or Thymine, S represents Guanine or Cytosine, W represents Adenine or Thymine, H represents a nucleotide other than Guanine, B represents a nucleotide other than Adenine, V represents a nucleotide other than Thymine, D represents a nucleotide other than Cytosine and N represents any nucleotide residue.
The designation of amino acid residues referred to herein, as recommended by the IUPAC-IUB
Biochemical Nomenclature Commission, are listed in Table 1.

Amino Acid Three-letter code One-letter code Alanine Ala A

Arginine Arg R

Asparagine Asn N

Aspartic acid Asp D

Cysteine Cys C

10Glutamine Gln Q

Glutamic acid Glu E

Glycine Gly G

Histidine His H

Isoleucine Ile I

15Leucine Leu L

Lysine Lys K

Methionine Met M

Phenylalanine Phe F

Proline Pro P

20Serine Ser S

Threonine Thr T

Tryptophan Trp . W

Tyrosine Tyr Y

Valine Val V

25AspartatelAsparagine Baa B

GlutamatelGlutamine Zaa Z

Any amino acid Xaa X

_Q_ BACKGROUND TO THE INVENTION
Improvements in recombinant DNA technology have produced dramatic changes to the agricultural industry, in particular the approaches taken to improve crop productivity.
A major concern is the effect of plant pests, such as plant fungal pathogens, on productivity. Plant pathogens such as fungi, rusts, nematodes, viruses and bacteria invade a wide range of food, fibre and ornamental plants, causing damage to different plant tissues reducing productivity.
Plant fungal pathogens, in particular rust fungi, represent an especially significant problem amongst broadacre crops such as legume and cereal grains.
Biotechnology offers considerable scope for addressing this problem, by introducing recombinant genes into plants that either kill or disable a fungal pathogen, or restrict a fungal pathogen to a limited zone of infection, thereby preventing significant deterioration of an economically-important crop.
An interaction between a rust pathogen and a plant host may be classed as either "resistant" or "susceptible" depending on how the fungal infection proceeds.
In a resistant interaction, infection by a fungal pathogen produces a "plant hypersensitive response" (Marineau et al., 1987; Dixon and Lamb, 1990} resulting in cell death to limit spread of the fungus. During the hypersensitive response, the expression of several infection-related genes, for example genes encoding phytoalexins, antimicrobial agents and pathogenesis-related (PR) proteins, is switched on. In contrast, a susceptible interaction involves no hypersensitive cell death and the infection alters host cell gene expression in such a way as to provide gene products that are essential for the biotrophic growth of an obligate plant pathogen. Additionally, host:pathogen interactions may be partially susceptible, in which case there may be a delay of several days in the appearance of PR proteins and the level of their expression is reduced compared to fully hypersensitive responses. It is also possible that both resistant and susceptible interactions may operate to varying degrees in any given pathogenic infection.

WO 99/4511$ PCT/AU99/00130 Although several studies have concentrated on characterising host:pathogen interactions during fungal infection of plants at the genetic level, in particular in a resistant interaction between the host plant and fungal pathogen, the identification of host-derived genes capable of conferring pathogen-resistance on plants has not been a straightforward procedure. In particular, although fungal pathogens are a major infections agent of monocotyledonous broadacre crop plants, the generation time and genome organisation of these plants has rendered the isolation of such sequences difficult, even in fight of well-characterised genetic systems. Moreover, those skilled in the art would be aware of the difficulties associated with the extrapolation of mere genetic linkage data to physical distances between loci in the complex genome of a monocotyledonous plant.
SUMMARY OF THE INVENTION
In accordance with the present invention, genetic sequences conferring resistance to 1S a plant pathogen, such as a plant fungal, nematode, viral or bacterial pathogen, have been cloned from Zea mays and Hordeum spp. The cloning of these sequences permits the generation of transgenic plants with de novo, improved or otherwise enhanced pathogen resistance, in particular resistance against fungal pathogens, such as rusts, and more particularly, rusts of the genera Puccinia or related rusts capable of infecting cereal crop plants.
The nucleotide sequences exempl~ed herein provide the means by which variant gene sequences may be isolated which confer resistance to a wide range of plant pathogens such as viruses, fungi, bacteria and nematodes and more particularly wheat streak mosaic virus, barley stem rust, Pyricularia ssp. and Xanthomonas ssp.
The present invention also permits the screening through genetic or immunological means, similar pathogen resistance genes in other plants for use in developing or enhancing pathogen resistance in commercially and economically important species.
Accordingly, one aspect of the present invention provides an isolated nucleic acid molecule comprising an Rp1-D nucleotide sequence or Rpg1 nucleotide sequence or a variant thereof which encodes a protein or derivative thereof, which confers, enhances, or otherwise facilitates resistance to a pathogen in a plant.
In another embodiment, the present invention provides an isolated nucleic acid molecule derived from a plant and comprising an Rp1-D nucleotide sequence or an Rpg1 nucleotide sequence or a variant thereof which:
(i) encodes or is complementary to a sequence encoding a polypeptide of plant origin which confers, enhances, or otherwise facilitates pathogen resistance in a plant; and (ii) has at least about 60% nucleotide sequence identity to any one or more of the nucleotide sequences set forth in SEQ ID NOS: <400>1 or <400>3 or <400>5 or <400>7, or SEQ ID Nos:<400>64 to <400>70, or a part thereof.
In yet another embodiment, the present invention provides an isolated nucleic acid molecule derived from a plant and comprising an Rp1-D nucleotide sequence or an Rpg1 nucleotide sequence or a variant thereof which:
(i) encodes or is complementary to a sequence encoding a polypeptide of plant origin which confers, enhances, or otherwise facilitates pathogen resistance in a plant; and (ii) hybridises under at least low stringency conditions to the Rp1-D
nucleotide sequence set forth in SEQ ID NOS: <400>1 or <400>3 or <400>5 or <400>7 or SEQ ID Nos:<400>64 to <400>70 or to a complementary strand thereof.
in yet another embodiment, the invention provides an isolated nucleic acid molecule which is substantially the same as any one or more of the Rp1-D nucleotide sequences set forth in SEQ ID NOS: <400>1 or <400>3 or the Rpg1 nucleotide sequences set forth in SEQ ID NOS:<400>5 or <400>7, or the hrpl sequences set forth in SEQ
ID
Nos:<400>64 to <400>70 or which is at least about 60% identical thereto.

Another aspect of the invention provides a genetic construct comprising an Rp1-D
nucleotide sequence or an Rpg1 nucleotide sequence or a variant thereof derived from a plant and encoding a protein or derivative thereof, which confers, enhances, or otherwise facilitates resistance to a pathogen in a plant or a compiementary nucleotide sequence thereto. According to one embodiment, the Rp1-D nucleotide sequence or Rpg1 nucleotide sequence is operably linked to a promoter sequence, thereby regulating expression of said nucleotide sequence in a eukaryotic cell, for example a plant cell, or a prokaryotic cell.
The invention extends to the recombinant Rp1-D polypeptide product or Rpg1 polypeptide product of said genetic construct.
The present invention also provides an oligonucleotide molecule of at least 10 nucleotides in length capable of hybridising under low stringency conditions to part of the Rp1-D nucleotide sequence set forth in SEQ ID NOS: <400>1 or <400>3 or the Rpg1 nucleotide sequences set forth in SEQ ID NOS:<400>5 or <400>7, or the hrp1 nucleotide sequences set forth in SEQ ID Nos:<400>fi4 to <400>70 or to a complementary nucleotide sequence thereto.
The Rp1-D nucleotide sequence andlor hrpl nucleotide sequence andlor Rpg1 nucleotide sequence andlor oligonucleotides comprising same or hybridising thereto are useful in the isolation of pathogen resistance or pathogen resistance-like genetic sequences from other plants, including those genetic sequences which are involved in resistance against both fungal pathogens and non-fungal pathogens, using hybridisation andlor PCR-based approaches.
Accordingly, there is provided a method of identifying a pathogen resistance genetic sequence or pathogen resistance-like genetic sequence which method comprises contacting genomic DNA, or mRNA, or cDNA, or parts, or fragments thereof, or a source thereof with an Rp1-D nucleotide sequence or with an Rpg1 nucleotide sequence or with an hrpl nucleotide sequence or a variant thereof which encodes a -$_ polypeptide which confers, enhances or otherwise facilitates pathogen resistance, or a part thereof, or a complementary nucleotide sequence thereto, and then detecting said hybridisation.
There is also provided a method of identifying a pathogen resistance genetic sequence or a pathogen resistance-like genetic sequence in a plant cell, which method comprises contacting genomic DNA, mRNA, or cDNA from said plant with one or more oligonucleotide molecules derived from an Rp9-D nucleotide sequence or an hrp1 nucleotide sequence or an Rpg9 nucleotide sequence or a variant thereof which encodes a polypeptide which confers, enhances or otherwise facilitates pathogen resistance, or a part thereof, or a complementary nucleotide sequence thereto, for a period of time and under conditions sufficient to form a double-stranded nucleic acid molecule and amplifying copies of the said genetic sequence in a polymerase chain reaction.
In another aspect, this invention also provides an isolated Rp1-D polypeptide or an Rpg1 polypeptide or a functional mutant, derivative part, fragment, or analogue of said polypeptide which confers, enhances, or otherwise facilitates resistance to a pathogen in a plant cell.
The present invention extends to a synthetic peptide comprising at least 10 contiguous amino acids of the Rp1-D amino acid sequence set forth in SEQ ID NO: <400>2 or SEQ ID NO: <400>4 or the Rpg1 amino acid sequence set forth in SEQ ID NO:
<400>6 or SEQ ID NO: <400>8 or having at least about 60% identity to all or a part thereof.
The polypeptide and synthetic peptides of the present invention may be used to generate specific immuno-interactive molecules. Accordingly, the present invention also contemplates an antibody that binds to an Rp1-D polypeptide or Rpg1 polypeptide which confers, enhances or otherwise facilitates resistance to a pathogen in a plant or an antigenic or immunogenic epitope thereof, wherein said polypeptide or epitope further comprises an amino acid sequence which is substantially the same as the amino acid sequence set forth in SEQ ID NO: <400>2 or SEQ ID NO: <400>4 or SEQ
ID NO: <400>6 or SEQ ID NO: <400>8 or is at least about 60% identical to all or a part thereof.
In yet another aspect of the present invention, there is provided a method of identifying a pathogen resistance gene product or pathogen resistance-like gene product in a plant cell, which method comprises contacting an antibody which is speciftc for an Rp1-D polypeptide or an Rpg1 polypeptide or an antigenic fragment or epitope thereof with an antigen from said plant for a period of time and under conditions sufficient to form an antibody-antigen complex and measuring the amount of said antibody-antigen complex formed.
Notwithstanding that the genetic sequences described herein are derived from plants by virtue of their fungal-resistance properties, the genetic sequences of the present invention are also useful for the generation of plants with generally-enhanced pathogen resistance characteristics, in particular resistance against a fungal, nematodes, viral or bacterial pathogen. Accordingly, there is also provided a plant carrying a non-endogenous Rp1-D nucleotide sequence or a non-endogenous Rpg1 nucleotide sequence or a variant thereof which when expressed in said plant confers, enhances, or otherwise facilitates pathogen resistance in said plant. The present invention extends to the progeny derived from said plant.
BRIEF DESCRIPTION OF THE DRAWINGS
Figure 1 is a graphical representation showing the Rp1 region of maize chromosome 10S showing the linkage relationships of genes for resistance against rusts (Rp), the Rp1-D gene and several RFLP or molecular markers.
Figure 2 is a copy of a photographic representation of Southern blot hybridisation of Ncol and Bg111 (panel B) restriction digests of DNA isolated from maize lines carrying different alleles of the Rp locus including Rp1-D. The Rp alleles present in each lane of panel A are as follows: Lane 1, rp-R168; Lane 2, Rp1 A; Lane 3, Rp1-B; Lane 4, blank; Lane 5, Rp1-C; Lane 6, Rp1-D; Lane 7, Rp1-F; Lane 8, Rp1-J, Lane 9, Rp1-M;
Lane 10, Rp1-I; Lane 11, Rp1-K; Lane 12, Rp1-C-K; Lane 13, Rp1-G; Lane 14, Rp1-G-l, Lane 15, Rp1-G-5; Lane 16, Rp1-G-F J; Lane 17, RpS; Lane 18, Rp1-Td; Lane 19, Rp5-C; Lane 20, Rp5-M; Lane 21, RpG-M; Lane 22, Rp1-D-13-2; Lane 23, Rp1-D13;
Lane 24, rp-W22; Lane 25, rp-a,sh2. The Rp1 alleles present in each lane of panel B
are as follows: Lane 1, rp-R168; Lane 2, Rp1 A; Lane 3, Rp1-B; Lane 4, Rp1-C;
Lane 5, Rp1-D; Lane 6, Rp1-F; Lane 7, Rp1-J, Lane 8, Rp1-M; Lane 9, Rp1-I; Lane 10, Rp1-K; Lane 11, Rp1-C-K; Lane 12, Rp1-G; Lane 13, Rp1-G-l, Lane 14, Rp1-G-5; Lane 15, Rp1-G-F J; Lane 16, RpS; Lane 17, Rp1-Td; Lane 18, Rp5-C; Lane 19, Rp5-M; Lane 20, RpG-M; Lane 21, Rp1-D-13-2; Lane 22, Rp1-D13; Lane 23, rp-I~V"12; Lane 24, rp-a,sh2. The filters were probed with the Rp1-D probe.
Figure 3 is a copy of a photographic representation of a Southern blot hybridisation of barley DNA isolated from individuals of a segregating family, digested with Drat and probed with the RpD-1 probe to show cross hybridisation and linkage with the Rpg1 rust-resistance locus in barley specifying resistance to barley rust, P.
graminis. Rpg1 = rust resistance; rpgl = rust susceptible. Arrows indicate the position of DNA bands which co-segregate with rust resistance.
Figure 4A is a representation of an amino acid sequence alignment of the maize Rp1-D and barley Rpg1-2 and barley Rpg1-13 polypeptides.
Figure 4B is a diagrammatic representation of the Rpg1-2 and Rp1-D
polypeptides, illustrating the similarity in amino acid sequences thereof. Figure 4B-I shows the relative location of kinase-1 a (p-loop) and kinase-2a and leucine-rich repeat (LRR) regions of these polypeptides. The overall percentages identity and similarity are indicated at the bottom of the Figure. Figure 4B- II shows the Kyte-Dolittle hydropathy plots of the Rpg1-2 polypeptide (top) and Rp1-D polypeptide (lower).
Figure 5 is a representation of a nucleotide sequence alignment between the coding regions of the maize Rp1-D gene and the barley Rpg1-2 and barley Rpg1-13 alleles.
Figure 6 is a copy of a photographic representation of a Southern blot hybridization of lUcol (left) or Bg111 (right) digested DNA from various grass species detected using the probe from the Rp1-D gene. The Figure shows cross hybridisation of the Rp1-D
gene sequence with sorghum (Sorghum vulgare; lane 1); pearl millet (Panicum mileraceum;
lane 2); maize (Zea mays; lane 3); sugar cane (Saccharum o~cinale; lane 4);
barley (Hordeum vulgare cv Morex; lane 5); wheat (Trificum aestivum cv Katyil; lane 6); oats (Avena safiva cv Marbo; lane 7); Triticum tauschii (lane 8); and rice (Oryza sativa cv Fuji; lane 9).
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS
One aspect of the present invention comprises an isolated nucleic acid molecule comprising an Rp1-D nucleotide sequence or an Rpg1 nucleotide sequence or a variant thereof which encodes a polypeptide which confers, enhances or otherwise facilitates resistance to a pathogen in a plant or a complementary nucleotide sequence thereto.
As used herein, the term "Rp1-D nucleotide sequence" shall be taken to refer to a nucleotide sequence that is the same as the maize Rp1-D gene set forth herein, or a fragment comprising at least about 20-30 contiguous nucleotide residues thereof or a complementary nucleotide sequence thereto.
As used herein, the term "Rpg1 nucleotide sequence" shall be taken to refer to a nucleotide sequence that is the same as the barley Rpg1-2 andlor barley Rpg1-alleles set forth herein, or a fragment comprising at least about 20-30 contiguous nucleotide residues thereof or a complementary nucleotide sequence thereto.
Hereinafter the term "pathogen resistance gene" or "pathogen resistance-like gene", or similar term shall be used to define a nucleic acid molecule which upon expression confers, enhances, or otherwise facilitates resistance of a cell andlor organism to one or more plant pathogens. The term "pathogen resistance gene" further defines a nucleic acid molecule which upon expression confers, enhances, or otherwise facilitates resistance to one or more plant pathogens selected from the list comprising fungal pathogens such as rusts, viral pathogens, bacterial pathogens and parasites such as nematodes. Particularly preferred pathogen resistance genes are those which are capable of conferring resistance against fungal or viral pathogens wherein said gene is a variant of the maize Rp1-D nucleotide sequence or a variant of the barley Rpg1 nucleotide sequences exemplified herein.
As used herein, the term "variant of an Rp1-D nucleotide sequence" or "Rp1-D
variant"
or similar term shall be taken to refer to any pathogen resistance gene derived from a plant wherein said gene comprises a nucleotide sequence which is at least about fi0%
identical to the Zea mays Rp1-D gene exemplified herein including the hrpl nucleotide sequences set forth as SEQ ID Nos:<400>64 to <400>70 exempl~ed herein; or a fragment thereof comprising at least about 30 nucleotides in length; or a pathogen resistance gene or fragment that hybridises to the Zea mays Rp1-D nucleotide sequence gene exemplified herein, or to a complementary sequence thereto, under at least low stringency hybridisation conditions. An Rp1-D variant may include an allelic variant of the specific Rp1-D gene sequence exemplified herein, in which case said allelic variant exhibits similar biological activity (i.e. against a similar or related pathogen). Alternatively, an Rp1-D variant may be derived from Zea mays or another plant species and exhibit different pathogen resistance characteristics to the Zea mays Rp1-D gene exemplified herein, for example a functionally-distinct Rp1 allele or other related Rp allele.
Similarly, the term "variant of an Rpg1 nucleotide sequence" or "Rpg1 variant"
or similar term shall be taken to refer to any pathogen resistance gene derived from a plant wherein said gene comprises a nucleotide sequence which is at least about 60%
identical to the barley Rpg1-2 or barley Rpg1-13 alleles exemplified herein;
or a fragment thereof comprising at least about 30 nucleotides in length; or a pathogen resistance gene or fragment that hybridises to the barley Rpg1-12 or barley Rpg1-13 alleles exemplified herein, or to a complementary sequence thereto, under at least low stringency hybridisation conditions. An Rpg1 variant may include an allelic variant of the specific Rpg1-2 andlor Rpg1-13 allelic gene sequences exemplified herein, in which case said allelic variant exhibits similar biological activity (i.e.
against a similar or related pathogen). Alternatively, an Rpg1 variant may be derived from Horcleum spp. or another plant species and exhibit different pathogen resistance characteristics to the Hordeum spp. Rpg1-2 and Rpg1-13 alleles exemplified herein, for example a functionally-distinct Rpg1 allele or other related Rpg1 allele.
The term "Rp1-D variant polypeptide" or similar shall be taken to refer to the polypeptide product of an Rp1-D variant gene as hereinbefore defined, in particular a polypeptide produced of the hrp1 nucleotide sequence set forth in any one or more of SEQ ID Nos:<400>64 to <400>70, or an immunological or functional homologue, analogue or derivative thereof.
The term "Rpg1 variant polypeptide" or similar shall be taken to refer to the polypeptide product of an Rpg1 variant gene as hereinbefore defined or an immunological or functional homologue, analogue or derivative thereof.
Preferably, Rp1-D and Rpg1 variants are isolated from broadacre crop plants and more preferably from monocotyledonous broadacre crop plants such as cereals and grasses, for example Zea mays, pearl millet, sugar cane, wheat, barley, rye, rice, oats, triticale, sorghum and ryegrass.
In particular, the present inventors have isolated nucleic acid molecules comprising one or more Rp1-D andlor hrpl alleles and/or Rpg1 alleles which confer resistance to the rusts Puccinia sorghi (Rp1-D) and barley stem rust (Rpg1). The maize Rp1-D
allele has been isolated from transposon-tagged fines of Zea mays. The isolated Rp1-D
allele exemplified herein has been used as a hybridisation probe, to isolate the maize hrpl nucleotide sequences and barley Rpg1 nucleotide sequences exemplified herein.
Accordingly, the maize hrpl-d1, hrpl-d2, hrpl-d3, hrpl-d4, hrp1-d5 and hrpl-d6 and hrp1-cin4 nucleotide sequences and the barley Rpg1-2 and Rpg1-13 nucleotide sequences exemplified herein fall within the scope of the term "Rp1-d variant"
as defined herein. Similarly, the maize Rp1-D nucleotide sequence and maize hrp1-d1 to hrpl-d6 and hrpl-cin4 sequences exemplified herein is an "Rpg1 variant" as defined herein.
The inventors have also demonstrated the utility of the subject Rp1-D allele for the identification andlor characterisation of Rp1-D variants derived from other plant species, including allelic and non-allelic variants of the Zea mat's Rp1-D
nucleotide sequence in maize and other species.
Accordingly, particularly preferred Rp1-D variants contemplated herein include nucleotide sequences of the barley Rpg1 gene family and other alleles of the Rp1 gene family derived from a broadacre crop plant selected from the list comprising barley, pearl millet, maize, sugar case, wheat, oats, Triticum tauschii and rice.
In an even more particularly preferred embodiment, the pathogen-resistance gene of the present invention further comprises a sequence of nucleotides which is at least about 60% identical to at least about 30 contiguous nucleotides of the Zea mat's Rp1-D
nucleotide sequence set forth in SEQ ID NOS: <400>1 or <400>3 or the Zea mat's hrpl nucleotide sequences set forth in SEQ ID NOS:<400>64 to <400>70 or a complementary sequence thereto, or the Hordeum spp. Rpg1 nucleotide sequences set forth in SEQ ID NOS: <400>5 or <400>7 or a complementary sequence thereto.
For the purposes of nomenclature, the sequences shown in SEQ ID NOS: <400>1 and <400>3 relate to a Rp1-D resistance allele derived from Zea mat's which controls resistance to the rust pathogen Puccinia sorghi in a resistant interaction.
More particularly, SEQ ID NO: <400>1 describes a genomic clone isolated from Zea mat's, containing the Rp1-D allele 13. The nucleotide sequence set forth in SEQ ID
NO:
<400>3 is an Rp1-D cDNA clone which was isolated from Zea mat's.

WO 99/45118 . PCTIAU99/00130 The amino acid sequence encoded by the genomic Rp1-D gene is set forth in SEQ
ID
NO: <400>2. The amino acid sequence of the complete Rp1-D polypeptide encoded by the Rp1-D cDNA sequence is presented in SEQ ID NO: <400>4.
The nucleotide sequences shown in SEQ ID NOS: <400>5 and <400>7 relate to the Rpg1-2 and Rpg1-13 resistance alleles derived from Hondeum spp. which controls resistance to barley stem rust in a resistant interaction. More particularly, SEQ ID NO:
<400>5 describes a genomic clone isolated from barley, containing an approximately 8.2 kb EcoRl-to-San fragment of the Rpg1-2 allele. The nucleotide sequence set forth in SEQ ID NO: <400>7 corresponds to the barley Rpg1-13 allele.
The amino acid sequence encoded by the genomic Rpg1-2 gene is set forth in SEQ
ID NO: <400>6. The amino acid sequence of the Rpg1-13 pofypeptide encoded by the Rpg1-13 allele is presented in SEQ ID NO: <400>8.
The present invention clearly extends to any one or more of the above-mentioned isolated nucleic acids or functional fragments, homologues, analogue or derivatives thereof, when integrated into a plant genome and to propagated plants containing one or more of said nucleic acid molecules, fragments, homologues, analogues or derivatives.
The Rp1-D variant or Rpg1 variant of the invention may confer resistance to a diverse range of fungal pathogens including stem rusts, leaf rust, yellow rust and crown rust, or to one or more nematode, viral or bacterial pathogens, amongst others. In a particularly preferred embodiment, the Rp1-D variant or Rpg1 variant of the present invention is a rust-resistance gene or a fragment or derivative thereof. More preferably, the Rp1-D variant or Rpg1 variant of the present invention is capable of conferring resistance against Puccinia andlor against barley stem rust in a plant.
Reference herein to a "gene" is to be taken in its broadest context and includes:
(i) a classical genomic gene consisting of a coding region optionally together with transcriptional andlor translational regulatory sequences and a coding region with or without non-translated sequences (i.e. introns, 5'- and 3'-untranslated sequences); or (ii) mRNA or cDNA corresponding to the coding regions (i.e. exons) and optionally 5'- and 3'- untranslated sequences of the gene.
The term "gene" is also used to describe a synthetic or fusion molecule, or derivative which encodes, or is complementary to a molecule which encodes, all or part of a functional product. A functional product is one which confers, enhances or otherwise facilitates resistance of a cell to a fungal pathogen.
Genetic analysis indicates that specific interactions may occur between resistance genes and gene products of the pathogen. Although not intending to limit the present invention to any one theory or mode of action, it is proposed that the genetic sequences of the present invention may control host range via specific recognition of the gene products of the pathogen pest, in a "gene-for-gene" interaction that is understood by one normally skilled in the art. Accordingly, the genetic sequences are useful in increasing the range of resistance of plants to distinct pathogen pests, by providing de novo the required pathogen resistance gene, or being introduced together with the corresponding pathogen gene or genes, on, for example, a single genetic cassette. Accordingly, these aspects of the invention are covered by the expression "conferring, improving, or otherwise enhancing pathogen resistance" or other similar expression.
The present invention clearly extends to any Rp9-D variant or Rpg1 variant which comprises a pathogen resistance gene and any functional gene, mutant, derivative, part, fragment, homologue or analogue thereof or non-functional molecule which is at least useful as, for example, a genetic probe, or primer sequence in the enzymatic or chemical synthesis of said gene, or in the generation of immunologically interactive recombinant molecules.

In a particularly preferred embodiment, the Rp1-D nucleotide sequences andlor the Rpg1 nucleotide sequences of the present invention (or like genetic sequences) are employed to identify and isolate similar genes, or pathogen resistance-like genes from other plants. The present invention extends to the use of said genetic sequence, or a part thereof to detect polymorphisms of a pathogen resistance genetic sequence or pathogen resistance-like genetic sequence.
Allelic and other variants of the Rp1-D nucleotide sequences andlor hrpl nucleotide sequences andlor Rpg1 nucleotide sequences may be detected andlor isolated using the Rp1-D andlor Rpg1 nucleotide sequences exemplified herein by any means known to those skilled in the art. In particular, the nucleotide sequences disclosed herein provide the means for isolation andlor detection of related nucleotide sequences, for example other Rp1 alleles and Rpg1 alleles, using standard nucleic acid hybridisation or amplification approaches, without undue experimentation.
Those skilled in the art will be aware that a functional assignment of an Rp1-D variant andlor an Rpg1 variant identified by hybridisation andlor amplification approaches may further require genetic linkage or cosegregation data to demonstrate that said allele is closely associated with a particular resistance phenotype. Those skilled in the art will be readily capable of performing such embodiments without undue experimentation.
Accordingly, in a further aspect of the invention, there is provided an oligonucleotide molecule of at least about 10 nucleotides in length and more preferably at least about 25-30 nucleotides in length capable of hybridising under low stringency conditions to part of the nucleotide sequence, or to a complement of any one or more of the nucleotide sequences set forth in SEQ ID NOS: <400>1 and/or <400>3 and/or <400>5 and/or <400>7 and/or <400>64 to <400>70.
A further aspect of the present invention contemplates an isolated nucleic acid molecule which encodes a protein that confers or otherwise facilitates pathogen resistance in a plant and which is capable of hybridising under at least low stringency WO 99!45118 PCT/AU99/00130 conditions to the nucleic acid molecule set forth in any one or more of SEQ ID
NOS:
<400>1 and/or <400>3 andlor <400>5 and/or <400>7 andlor any one of SEQ ID
NOS:<400>64 to <400>70 or to a derivative, homologue or analogue thereof.
Preferably, said nucleic acid molecule is an Rp1-D variant or an Rpg1 variant.
For the purposes of defining the level of stringency, a low stringency is defined herein as being a hybridisation and/or a wash carried out in 6xSSC buffer, 0.1 %
(wlv) SDS
at 28°C. Generally, the stringency is increased by reducing the concentration of SSC
buffer, andlor increasing the concentration of SDS andlor increasing the temperature of the hybridisation andlor wash. Conditions for hybridisations and washes are well understood by one normally skilled in the art. For the purposes of clarification of parameters affecting hybridisation between nucleic acid molecules, reference can conveniently be made to pages 2.10.8 to 2.10.16. of Ausubel et al. (1987), which is herein incorporated by reference.
Preferably, variants of the Rp1-D andlor Rpg1 alleles exemplified herein are isolated by hybridisation under medium or more preferably, under high stringency conditions, to a probe which comprises at least about 30 contiguous nucleotides derived from SEQ
ID NOS: <400>1 andlor <400>3 and/or <400>5 and/or <400>7 andlor any one of SEQ
ID NOS:<400>64 to <400>70 or a complement thereof.
The Rp1-D andlor Rpg1 variant gene sequences thus isolated may be modified by standard recombinant techniques. Generally, a pathogen resistance gene may be subjected to mutagenesis to produce single or multiple nucleotide substitutions, deletions andlor additions. Nucleotide insertional derivatives of the pathogen resistance gene of the present invention include 5' and 3' terminal fusions as well as intra-sequence insertions of single or multiple nucleotides. insertional nucleotide sequence variants are those in which one or more nucleotides are introduced into a predetermined site in the nucleotide sequence although random insertion is also possible with suitable screening of the resulting product. Deletional variants are characterised by the removal of one or more nucleotides from the sequence.

Substitutional nucleotide variants are those in which at least one nucleotide in the sequence has been removed and a different nucleotide inserted in its place.
Such a substitution may be "silent" in that the substitution does not change the amino acid defined by the codon. Alternatively, substituents are designed to alter one amino acid for another similar acting amino acid, or amino acid of like charge, polarity, or hydrophobicity.
Another aspect of the present invention is directed to a nucleic acid molecule which comprises a sequence of nucleotides corresponding or complementary to the nucleotide sequence set forth in any one of SEQ ID NOS: <400>1 or <400>3 or <400>5 or <400>7 andlor SEQ ID NOS:<400>64 to <400>70 or a homologue, analogue or derivative thereof having at least about 60% identity thereto, wherein said nucleic acid molecule encodes a protein which confers, enhances, or otherwise facilitates resistance to a pathogen in a plant.
Preferably, the percentage similaritty to any one of said sequences set forth is at least about 70%. Even more preferably, the percentage similarity is at least 80-90%, including at least 91 % or 93% or 95%.
In determining whether or not two nucleotide sequences fall within these percentage limits, those skilled in the art will be aware that it is necessary to conduct a side-by-side comparison or multiple alignment of sequences. In such comparisons or alignments, differences may arise in the positioning of non-identical residues, depending upon the algorithm used to perform the alignment. In the present context, reference to a percentage identity between two or more nucleotide sequences shall be taken to refer to the number of identical residues between said sequences as determined using any standard algorithm known to those skilled in the art. For example, nucleotide sequences may be aligned and their identity calculated using the BESTF1T
programme or other appropriate programme of the Computer Genetics Group, Inc., University Research Park, Madison, Wisconsin, United States of America (Devereaux ef al, 1984). Alternatively or in addition, wherein two or more nucleotide sequences are being compared, the ClustalW programme of Thompson et a! (1994) is used, as is the case for data presented herein as Figure 5.
For the present purpose, "homologues" of a nucleotide sequence shall be taken to refer to an isolated nucleic acid molecule which is substantially the same as the nucleic acid molecule of the present invention or its complementary nucleotide sequence, notwithstanding the occurrence within said sequence, of one or more nucleotide substitutions, insertions, deletions, or rearrangements.
"Analogues" of a nucleotide sequence set forth herein shall be taken to refer to an isolated nucleic acid molecule which is substantially the same as a nucleic acid molecule of the present invention or its complementary nucleotide sequence, notwithstanding the occurrence of any non-nucleotide constituents not normally present in said isolated nucleic acid molecule, for example carbohydrates, radiochemicals including radionucleotides, reporter molecules such as, but not limited to DIG, alkaline phosphatase or horseradish peroxidase, amongst others.
"Derivatives" of a nucleotide sequence set forth herein shall be taken to refer to any isolated nucleic acid molecule which contains significant sequence identity to said sequence or a part thereof. Generally, the nucleotide sequence of the present invention may be subjected to mutagenesis to produce single or multiple nucleotide substitutions, deletions andlor insertions. Nucleotide insertional derivatives of the nucleotide sequence of the present invention include 5' and 3' terminal fusions as well as intra-sequence insertions of single or multiple nucleotides or nucleotide analogues.
Insertional nucleotide sequence variants are those in which one or more nucleotides or nucleotide analogues are introduced into a predetermined site in the nucleotide sequence of said sequence, although random insertion is also possible with suitable screening of the resulting product being performed. Deletional variants are characterised by the removal of one or more nucleotides from the nucleotide sequence. Substitutional nucleotide variants are those in which at least one nucleotide in the sequence has been removed and a different nucleotide or nucleotide analogue inserted in its place.
Particularly preferred homologues, analogues or derivatives of the nucleotide sequences of the present invention include any one or more of the isolated nucleic acid molecules selected from the following:
(i) an isolated nucleic acid molecule which comprises a nucleotide sequence which is at least about 60% identical to any one of SEQ ID NOS:
<400>1 andlor <400>3 and/or <400>5 andlor <400>7 and/or SEQ ID
NOS:<400>64 to <400>70 or a complementary sequence thereto;
(ii) an isolated nucleic acid molecule which comprises a nucleotide sequence which is at least about 60% identical to at least about 30 contiguous nucleotides of SEQ ID NOS: <400>1 andlor <400>3 and/or <400>5 andlor <400>7 or any one of SEQ ID NOS:<400>64 to <400>70 or a complementary sequence thereto;
(iii) an isolated nucleic acid molecule which is capable of hybridising under at least low stringency conditions to at least 10 contiguous nucleotides, preferably at least about 25-30 contiguous nucleotides of SEQ ID NOS: <400>1 andlor <400>3 andlor <400>5 andlor <400>7 or any one of SEQ ID
NOS:<400>64 to <400>70 or a complementary sequence thereto;
(iv) an isolated nucleic acid molecule which comprises a sequence of nucleotides which encodes or is complementary to a sequence of nucleotides which encodes an Rp1-D variant polypeptide;
(v) an isolated nucleic acid molecule which comprises a sequence of nucleotides which encodes or is complementary to a sequence of nucleotides which encodes an Rpg1 variant polypeptide (vi) an isolated nucleic acid molecule which comprises a nucleotide sequence or is complementary to a nucleotide sequence which encodes a pathogen-resistance polypeptide which is at least about 60% identical at the amino acid sequence level to the amino acid sequence set forth in SEQ ID
NOS: <400>2 andlor <400>4 andlor <400>6 andlor <400>8; and (vii) an isolated nucleic acid molecule which comprises a nucleotide WO 99/45118 ~ PCT/AU99/00130 sequence or is complementary to a nucleotide sequence which encodes a pathogen-resistance polypeptide which at least comprises about 10 contiguous amino acids of the sequence set forth in SEQ ID NOS: <400>2 and/or <400>4 andlor <400>6 and/or <400>8.
A further aspect of the invention contemplates a method for identifying a pathogen resistance genetic sequence or pathogen resistance-like genetic sequence, said method comprising contacting genomic DNA, or mRNA, or cDNA, or parts, or fragments thereof, or a source thereof, with a hybridisation effective amount of a probe comprising an Rp1-D nucleotide sequence or an hrpl nucleotide sequence or an Rpg1 nucleotide sequence or a related pathogen resistance gene or a homologue, analogue or derivative thereof for a time and under conditions sufficient for hybridisation to occur and then detecting said hybridisation.
The pathogen resistance genetic sequence or like sequence may be in a recombinant form, in a virus particle, bacteriophage particle, yeast cell, animal cell, or a plant cell.
Preferably, said genetic sequence originates from a broadacre crop plant, in particular a monocotyledonous plant such as maize, barley, rye, oats, wheat, sorghum, triticale, ryegrass or rice and/or wild varieties andlor hybrids or derivatives and/or ancestral progenitors of same. In addition, the identified pathogen resistance genetic sequence or the probe may be bound to a support matrix, for example nylon, nitrocellulose, polyacrylamide, agarose, amongst others.
Preferably, the probe comprises the nucleotide sequence of the Zea mays Rp1-D
variant exemplified herein or the barley Rpg1 variant allele exemplified herein or a related sequence such as, but not limited to, an Rp1-D nucleotide sequence or an hrpl nucleotide sequence or an Rpg1 nucleotide sequence derived from another plant species, amongst others.
In a most preferred embodiment, the probe comprises the sequence of nucleotides set forth in any one of SEQ ID NOS: <400>1 or <400>3 or <400>5 or <400>7 or any one WO 99/45118 ~ PCT/AU99/00130 of SEQ 1D NOS:<400>64 to <400>70 or a homologue, derivative or analogue thereof or complementary sequence thereto.
Preferably, the probe is labelled with a reporter molecule capable of giving an identifiable signal (e.g. a radioisotope such as 32P or 35S or a biotinylated molecule).
An alternative method contemplated in the present invention involves hybridising a nucleic acid primer molecule of at feast 10 nucleotides in length derived from an Rp9-D
nucleotide sequence or an hrp9 nucleotide sequence or Rpg1 nucleotide sequence or a pathogen-resistance gene which is related thereto or a complementary sequence thereto to a nucleic acid "template molecule". Specific nucleic acid molecule copies of the template molecule are amplified enzymatically in a polymerase chain reaction, a technique that is well known to one skilled in the art and described for example by McPherson et al. (1991).
Preferably, the nucleic acid primer molecule or molecule effective in hybridisation is contained in an aqueous mixture of other nucleic acid primer molecules. More preferably, the nucleic acid primer molecule is in a substantially pure form.
In a particularly preferred embodiment, the nucleic acid primer molecule is derived from Zea mays, or other monocotyledonous plant such as barley or wheat. In a more preferred embodiment, the nucleic acid primer molecule comprises a nucleotide sequence of at least about 25-30 nucleotides in length derived from, orcontained within the nucleotide sequences of the sequences set forth in any one or more of SEQ ID NOS: <400>1 or <400>3 or <400>5 or <400>7 or SEQ ID NOS:<400>64 to <400>70 or a homologue, analogue or derivative thereof.
In a particularly preferred embodiment, the primer and/or probe described according to the preceding embodiments of the present invention comprise a nucleotide sequence set forth in any one of SEQ ID NOS:<400>34 to <400>63.
The nucleic acid template molecule may be in a recombinant form, in a virus particle, bacteriophage particle, yeast cell, animal cell, or a plant cell. Preferably, the related genetic sequence is derived from a monocotyledonous plant such as maize, wheat, barley, triticale, rye, oats, rice or ryegrass andlor wild varieties and/or hybrids or derivatives andlor ancestral progenitors of same.
A further aspect of the present invention is directed to a genetic construct which at least comprises an isolated Rp1-D nucleotide sequence or an isolated hrpl nucleotide sequence or an isolated Rpg1 nucleotide sequence or a homologue, analogue or derivative thereof.
Preferably, the Rp1-D nucleotide sequence shares at least about 60% identity to SEQ
ID NOS: <400>1 or <400>3 or is a functional derivative, part fragment, homologue, or analogue thereof.
Preferably, the Rpg1 nucleotide sequence shares at least about 60% identity to SEQ
ID NOS: <400>5 or <400>7 or is a functional derivative, part fragment, homologue, or analogue thereof.
Preferably, the hrp1 nucleotide sequence shares at least about 60% identity to any one of SEQ ID NOS:<400>64 to <400>70 or is a functional derivative, part, fragment, homologue or analogue thereof.
More preferably, the genetic construct comprises the entire open reading flame of an Rp?-D variant gene sequence or the hrpl gene sequence or the Rp1-D variant gene sequence or a fragment thereof which is at least useful as a probe to isolate related sequences.
Alternatively, the subject genetic construct may comprise a region of the open reading frame of an Rp1-D variant or hrp1 variant or Rp1-D variant, such as a region which encodes at least about 25 to 30 contiguous amino acids thereof, which is useful for the expression of a recombinant epitope of a pathogen-resistance polypeptide.

Accordingly, this embodiment of the present invention clearly extends to genetic constructs for the expression of fusion polypeptides comprising epitopes derived from different pathogen-resistance polypeptides, in particular, fusion polypeptides derived from the Rp1-D and/or Rpg1 variants described herein. Such fusion polypeptides may confer novel resistance phenotypes on transgenic plants in which they are expressed, such as being resistant to different fungal pathogens to the resistance conferred by the base polypeptides from which they are derived.
Those skilled in the art will be aware that recombinant epitopes of a pathogen-resistance polypeptide may also be useful as immunogens for the preparation of antibody molecules, such as polyclonal antibodies, monoclonal antibodies and fragments and immunoglobulin fractions thereof.
Additionally, recombinant Rp1-D andlor Rpg1 epitopes and/or hrp1 epitopes and Rp1-D and/or Rpg1 variant nucleotide sequences andlor hrp1 variant nucleotide sequences encoding same are useful in screening for the presence andlor expression of pathogen-resistance alleles in plants. The recombinant epitopes and nucleotide sequences may be selected such that they are specific for any particular Rp1-D
andlor Rpg1 variant allele and/or hrpl variant allele or alternatively, comprise amino acid sequences common to several different Rp1-D and/or Rpg1 andlor hrp1 variant poiypeptides.
For example, the present inventors have shown that the Rp1-D allele set forth in SEQ
ID NOS: <400>1 and <400>3 shares approximately 58% sequence identity with the corresponding region of the barley Rpg1-2 allele (SEQ ID NO:<400>5) and approximately 60% sequence identity with the corresponding region of the barley Rpg1-13 allele {SEQ ID NO:<400>7), whilst the barley Rpg1-2 and Rpg1-13 alleles are 74% identical overall, wherein higher sequence homology is present in gene regions which encode at least the kinase-1 a or p-loop motif core (amino acids 218 to 226 of SEQ ID NO:<400>2 or SEQ ID NO: <400>4), kinase-2a motif core {amino acids 296 to 302 of SEQ ID NO:<400>2 or SEQ ID NO: <400>4), CFL motif (amino acids 447 to 453 of SEQ ID NO:<400>2 or SEQ ID NO: <400>4) and WVAEG motif (amino acids 469 to 473 of SEQ ID NO:<400>2 or SEQ ID NO: <400>4) of the nucleotide binding sitelleucine-rich repeat (NBS-LRR) proteins encoded by this particular class of disease-resistance genes (i.e. the Rp1-D gene family).
These highly conserved motifs are particularly useful for the generation of irnmunologically interactive molecules which are capable of binding to an Rp1-D variant polypeptide or Rpg1 variant polypeptide. In an alternative embodiment, Rp1-D
andlor Rpg1 nucleotide sequences encoding these highly conserved motifs may be placed operably in connection with a suitable promoter sequence to facilitate the recombinant production of peptides comprising these motifs, optionally to facilitate subsequent antibody production thereto. Those skilled in the art will also be aware that peptides comprising these highly conserved motifs may be produced by synthetic means.
The present invention clearly extends to genetic constructs designed to assist expression of an Rp1-D variant polypeptide or Rpg1 variant polypeptide or hrp1 variant polypeptide which is capable of conferring, enhancing or facilitating pathogen resistance in a cell or to assist the expression of an immunological epitope or fragment thereof.
Generally, wherein expression of the Rp1-D variant polypeptide or hrp1 variant polypeptide or Rpg1 variant polypeptide or epitope or fragment thereof is desired, the genetic construct comprises in addition to the subject nucleic acid molecule, a promoter and optional other regulatory sequences that modulate, regulate or direct expression of the nucleic acid molecule encoding said polypeptide. The promoter may be the Rp1-D gene promoter, the Rpg1 gene promoter or a promoter from another genetic source. Preferably, however, the promoter is capable of expression in a plant cell, either constitutively or in response to infection by a fungal, nematode, viral or bacterial pathogen.
The Rp1-D variant nucleic acid molecule may be genomic DNA or cDNA and may correspond in sequence to the nucleotide sequence set forth in SEQ ID NOS:
<400>1 or <400>3 or <400>5 or <400>7 or <400>64 to <400>70 or a homologue, analogue or derivative thereof.
Reference herein to a "promoter" is to be taken in its broadest context and includes the transcriptional regulatory sequences of a classical eukaryotic genomic gene, including the TATA box which is required for accurate transcription initiation, with or without a CCAAT box sequence and additional regulatory elements (i.e. upstream activating sequences, enhancers and silencers) which alter gene expression in response to developmental andlor external stimuli, or in a tissue-specific manner. In the context of the present invention, the term "promoter" also includes the transcriptional regulatory sequences of a classical prokaryotic gene, in which case it may include a -35 box sequence and/or a -10 box transcriptional regulatory sequences.
In the present context, the term "promoter" is also used to describe a synthetic or fusion molecule, or derivative which confers, activates or enhances expression of said sense molecule in a cell Preferred promoters may contain additional copies of one or more specific regulatory elements, to further enhance expression andlor to alter the spatial expression andlor temporal expression of said sense molecule.
Placing a genetic sequence under the regulatory control of a promoter sequence means positioning said molecule such that expression is controlled by the promoter sequence. A promoter is usually, but not necessarily, positioned upstream or 5' of a nucleic acid molecule which it regulates. Furthermore, the regulatory elements comprising a promoter are usually positioned within 2 kb of the start site of transcription. In the construction of heterologous promoterlstructural gene combinations it is generally preferred to position the promoter at a distance from the gene transcription start site that is approximately the same as the distance between that promoter and the gene it controls in its natural setting, i.e., the gene from which the promoter is derived. As is known in the art, some variation in this distance can be accommodated without loss of promoter function. Similarly, the preferred positioning of a regulatory sequence element with respect to a heterologous gene to be placed under its control is defined by the positioning of the element in its natural setting, i.e., the genes from which it is derived. Again, as is known in the art, some variation in this distance can also occur.
Examples of preferred promoters suitable for use in genetic constructs of the present invention include promoters derived from the genes of viruses, yeasts, moulds, bacteria, insects, birds, mammals and plants which are capable of functioning in isolated plant cells and more particularly, monocotyledonous plant cells or whole organisms regenerated therefrom. The promoter may regulate the expression of the pathogen-resistance polypeptide constitutively, or differentially with respect to the tissue in which expression occurs or, with respect to the developmental stage at which expression occurs, or in response to external stimuli such as physiological stresses, pathogens, or metal ions, amongst others.
Examples of promoters include the CaMV 35S promoter, NOS promoter, octopine synthase (OCS) promoter, Arabidopsis thaliana SSU gene promoter, napin seed-specific promoter, P~ promoter, BK5-T imm promoter, lac promoter, tac promoter, phage lambda l~~ or l~ promoters, CMV promoter {U.S. Patent No. 5,168,062), T7 promoter, IacUV5 promoter, SV40 early promoter (U.S. Patent No. 5,118,627), late promoter {U.S. Patent No. 5,118,627), adenovirus promoter, baculovirus P10 or polyhedrin promoter (U.S. Patent NOS. 5,243,041, 5,242,687, 5,266,317, 4,745,051 and 5,169,784), and the like. In addition to the specific promoters identified herein, cellular promoters for so-called housekeeping genes are useful.
Preferred promoters according to this embodiment are those promoters which are capable of functioning in yeast, mould or plant cells. More preferably, promoters suitable for use according to this embodiment are capable of functioning in cells derived from monocotyledonous plants.
In a more preferred embodiment, the promoter may be derived from a genomic clone WO 99/45118 ~ PCTlAU99100130 encoding a Rp1-D variant polypeptide, preferably derived from the genomic gene set forth in SEQ ID NO: <400>1, more preferably from nucleotides 1 to about 1200 of SEQ
ID NO: <400>1 or a homologue, analogue or derivative thereof which is capable of conferring expression on a structural gene in a plant cell.
In an alternative preferred embodiment, the promoter may be derived from a genomic clone encoding an Rpg1 variant polypeptide, preferably derived from the Rpg1-2 allele set forth in SE4 ID NO: <400>5, more preferably from nucleotides 1 to about 3765 of SEQ ID NO: <400>5, or nucleotides from about position 1000 to about 3765 of SEQ
ID NO: <400>5, or a homologue, analogue or derivative thereof which is capable of conferring expression on a structural gene in a plant cell.
In an alternative preferred embodiment, the promoter may be derived the Rpg1-allele set forth in SEQ ID NO: <400>7, more preferably from nucleotides 1 to about 715 of SEQ ID NO: <400>7 or a homologue, analogue or derivative thereof which is capable of conferring expression on a structural gene in a plant cell.
In a more preferred embodiment, the promoter may be derived from a highly-expressed plant-expressible gene with a view to increasing expression of the pathogen resistance gene to which it is operably connected in the genetic construct.
The genetic construct of the invention may further comprise a tem~inator sequence and be introduced into a suitable host cell where it is capable of being expressed to produce a recombinant polypeptide gene product.
The term "terminator" refers to a DNA sequence at the end of a transcriptional unit which signals termination of transcription. Terminators are 3'-non-translated DNA
sequences containing a polyadenylation signal, which facilitates the addition of polyadenylate sequences to the 3'-end of a primary transcript. Terminators active in cells derived from viruses, yeasts, moulds, bacteria, insects, birds, mammals and plants are known and described in the literature. They may be isolated from bacteria, WO 99/45118 ~ PC'f/AU99/00130 fungi, viruses, animals andlor plants.
Examples of terminators particularly suitable for use in the genetic constructs of the present invention include the nopaline synthase (NOS) gene terminator of Agrobacterium tumefaciens, the terminator of the Cauliflower mosaic virus (CaM~ 35S
gene, the zein gene terminator from Zea mays, the Rubisco small subunit (SSU) gene terminator sequences, subclover stunt virus (SCS~ gene sequence terminators, any rho-independent E. coli terminator, amongst others.
Those skilled in the art will be aware of additional promoter sequences and terminator sequences which may be suitable for use in performing the invention. Such sequences may readily be used without any undue experimentation.
The genetic construct may further comprise a selectable marker gene or genes that are functional in a cell into which said genetic construct is introduced.
As used herein, the term "selectable marker gene" includes any gene which confers a phenotype on a cell in which it is expressed to facilitate the identification andlor selection of cells which are transfected or transformed with a genetic construct of the invention or a derivative thereof.
Suitable selectable marker genes contemplated herein include the ampicillin resistance (Amp), tetracycline resistance gene (Tt; }, bacterial kanamycin resistance gene (Kan~, phosphinothricin resistance gene, neomycin phosphotransferase gene (nptll), hygromycin resistance gene, (3-glucuronidase (GUS) gene, chloramphenicol acetyltransferase (CAT) gene and luciferase gene, amongst others.
Yet another aspect of the present invention provides for the expression of the subject pathogen-resistance genetic sequence in a suitable host (e.g. a prokaryote or eukaryote) cell, tissue, organ or organism to produce full length or non-full length recombinant pathogen resistance gene products. Preferably, the pathogen resistance WO 99/45118 ~ PCT/AU99I00130 gene product has an amino acid sequence that is identical to, or contained within the amino acid sequence set forth in any one of SEQ ID NOS: <400>2 or <400>4 or <400>6 or <400>8, or is at least about 60% identical to at least about 5 contiguous amino acids thereof.
In determining whether or not two amino acid sequences fall within these percentage limits, those skilled in the art will be aware that it is necessary to conduct a side-by-side comparison or mukiple alignment of sequences. In such comparisons or alignments, differences will arise in the positioning of non-identical residues, depending upon the algorithm used to perform the alignment. In the present context, reference to a percentage identity or similarity between two or more amino acid sequences shall be taken to refer to the number of identical and similar residues respectively, between said sequences as determined using any standard algorithm known to those skilled in the art. For example, amino acid sequence identities or similarities may be calculated using the GAP programme and/or aligned using the PILEUP programme of the Computer Genetics Group, Inc., University Research Park, Madison, Wisconsin, United States of America (Devereaux et al, 1984). The GAP programme utilizes the algorithm of Needleman and Wunsch (1970) to maximise the number of identicaUsimilar residues and to minimise the number and/or length of sequence gaps in the alignment.
Alternatively or in addition, wherein more than two amino acid sequences are being compared, the ClustalW programme of Thompson et al (1994) is used.
The present invention therefore provides a recombinant polypeptide which comprises an amino acid sequence which confers, enhances, or otherwise facilitates resistance to a pathogen in a plant cell, or a functional mutant, homologue, derivative, part, fragment, or analogue of said polypeptide.
In the present context, "homologues" of a poiypeptide refer to those polypeptides, enzymes or proteins which have similar properties as a polypeptide of the present invention, for example pathogen-resistance properties in relation to the infection of plants by a fungal pathogen, viral pathogen, nematode pathogen or bacterial pathogen;

amongst others, notwithstanding any amino acid substitutions, additions or deletions thereto.
Furthermore, amino acids may be replaced by other amino acids having similar properties, for example hydrophobicity, hydrophilicity, hydrophobic moment, antigenicity, propensity to form or break a-helical structures or ~-sheet structures or other conformational structures.
The present invention clearly extends to such amino acid variants, provided that such molecules still function as pathogen-resistance products in conferring, stimulating or otherwise enhancing resistance against a fungal, nematode, viral or bacterial pathogen or alternatively, comprise one or more B cell or T-cell linear or conformational epitopes capable of eliciting the production of antibodies which bind to said pathogen-resistance product or a fragment thereof.
Furthermore, a homologue may be isolated or derived from the same or another plant species as the Rp1-D or Rpg1 variant polypeptides exempl~ed herein. Preferred sources of homologues of the Zea mays Rp1-D variant set forth in SEQ ID NO:
<400>2 or SEQ ID NO: <400>4, or the barley Rpg1 variant poiypeptides set forth in SEQ
ID
NO: <400>6 or SEQ ID NO: <400>8, include any monocotyledonous plant species and more preferably pearl millet, maize, sugar cane, Trificum tauschii, rice, wheat, rye, oats, sorghum, triticale, ryegrass and barley, amongst others.
Furthermore, the amino acids of a homologous polypeptide may be replaced by other amino acids having similar properties, for example hydrophobicity, hydrophilicity, hydrophobic moment, charge or antigenicity, and so on.
"Analogues" encompass pathogen resistance polypeptides notwithstanding the occurrence of any non-naturally occurring amino acid analogues therein, such as D-stereoisomers and synthetic amino acid analogues.

The term "derivative" in relation to a pathogen resistance polypeptide, in particular an Rp1-D or Rpg1 variant polypeptide shall be taken to refer hereinafter to mutants, parts or fragments of a functional molecule. Derivatives include modified peptides in which ligands are attached to one or more of the amino acid residues contained therein, such as carbohydrates, enzymes, proteins, polypeptides or reporter molecules such as radionuclides or fluorescent compounds. Glycosylated, fluorescent, acylated or alkylated forms of the subject peptides are particularly contemplated by the present invention. Additionally, derivatives of a pathogen resistance polypeptide may comprise fragments or parts of an amino acid sequence disclosed herein and are within the scope of the invention, as are fusion polypeptides derived from two or more distinct Rp1-D and/or Rpg1 variant poiypeptides. Such a fusion poiypeptide may provide novel resistance characteristics compared to either Rp1-D and/or Rpg1 variant polypeptides from which it is derived.
Procedures for derivatizing peptides are well-known in the art.
Substitutions encompass amino acid alterations in which an amino acid is replaced with a different naturally-occurring or a non-conventional amino acid residue.
Such substitutions may be classified as "conservative", in which case an amino acid residue contained in a polypeptide is replaced with another naturally-occurring amino acid of similar character, for example Gly~-~Ala, Val~-~Ilet-~Leu, Asp~--~Glu, Lys~-~Arg, Asn~-~Gln or Phe~-~Trp~-~Tyr.
Substitutions contemplated herein may also be "non-conservative", in which an amino acid residue which is present in a polypeptide is substituted with an amino acid having different properties, such as a naturally-occurring amino acid from a different group (eg.
substituted a charged or hydrophobic amino acid with alanine), or alternatively, in which a naturally-occurring amino acid is substituted with a non-conventional amino acid.
Amino acid substitutions are typically of single residues, but may be of multiple residues, either clustered or dispersed.
Deletions and insertions may be made to the N-terminus, the C-terminus or be internal deletions or insertions.
The present invention clearly extends to a synthetic peptide fragment of a pathogen resistance gene product, the resistance gene product set forth in any one or more of SEQ ID NOS: <400>2 or <400>4 or <400>6 or <400>8 which may be useful in diagnostic applications or in the generation of antibody molecules.
According to this aspect, the present invention particularly provides a recombinant polypeptide product which is encoded by the Zea mays Rp1-D rust resistance gene or the barley Rpg9 rust resistance gene. This is done, however, with the understanding that the subject invention extends to a range of resistance gene products for rusts and other pathogens of monocotyledonous plants which are encompassed by the terms "Rp1-D variant polypeptide" and/or "Rpg1 variant polypeptide" andJor which satisfy the requirements of a homologue, analogue or derivative of the specific rust-resistance polypeptides exemplified herein.
In fact, the present invention extends to any recombinant poiypeptide product of a pathogen resistance gene characterised by said product having at least about 60%
similarly to at least about 5 contiguous amino acid residues of SEQ ID NO:
<400>2 or SEQ ID NO: <400>4 or SEQ ID NO: <400>6 or SEQ tD NO: <400>8, more preferably at least about 7 contiguous amino acids, even more preferably at least about 9 to 13 contiguous amino acids or at least about 15 to 1fi contiguous amino acids or at least about 19 to 20 contiguous amino acids of said amino acid sequences.
Preferably, the recombinant polypeptide of the invention further comprises one or more amino acid sequences selected from:
(a) CFLYCSL (SEQ ID NO: <400>9; equivalent to amino acids 447 to 453 of SEQ ID NO: <400>4); and more particularly, LQRCFLYCSLFPKGH (SEQ ID

NO: <400>10; equivalent to amino acids 444 to 458 of SEQ ID NO: <400>4) or a homologue, analogue or derivative thereof; and (b) WXAEG (SEQ ID NO: <400>11; equivalent to amino acids 469 to 473 of SEQ ID NO: <400>4}; and more particularly, ELVHLW(VIM)AEG (SEQ ID NO:
S <400>12; equivalent to amino acids 464 to 473 of SEQ ID NO: <400>4) or a homologue, analogue or derivative thereof.
More preferably, the recombinant polypeptide of the present invention further comprises an amino acid sequence motif characterised as a p-Loop, or kinase-1 a motif and having the sequence VGXGGXGKS (SEQ ID NO: <400>13; equivalent to amino acid residues 218 to 226 of SEQ ID NO: <400>4); and more particularly, YSGLAIVGXGGXGKSXLAQ (SEQ 1D NO: <400>14; equivalent to amino acid residues 212 to 230 of SEQ ID NO: <400>4) or a homologue, analogue or derivative thereof.
Even more preferably, the pathogen resistance gene product further contains a kinase-2 motif, having the sequence LLVLDDV (SEQ lD NO: <400>15; equivalent to amino acids 296 to 302 of SEQ ID NO: <400>4); and more particularIy~CFLLVLDDVWFE
(SEQ ID NO: <400>16; equivalent to amino acids 294 to 305 of SEQ ID NO:
<400>4), or a homologue, analogue or derivative thereof.
In a more particularly preferred embodiment, the recombinant polypeptide of the invention comprises one or more amino acid sequences selected from the list comprising:
(a) KYSGLAIVG {SEQ ID NO: <400>17);
(b) YSGLAIVGL (SEQ ID NO: <400>18);

(c} LGGMGKS (SEQ 1D NO: <400>19);

(d) GGMGKS (SEQ ID NO: <400>20);

(e) GGMGKST (SEQ ID NO: <400>21);

(f) GGMGKSTLA4 (SEQ ID NO: <400>22};

(g) GKSTLAQ (SEQ ID NO: <400>23);

(h) STLAQ (SEQ ID NO: <400>24);
(i) TLAQY (SEQ ID NO: <400>25);
{j) QKFLLVLDDVWFE (SEQ ID NO: <400>26);
(k) KFLLVLDDVWFEK (SEQ ID NO: <400>27);
(I) RLQRCFLYCSLFPKGH (SEQ ID NO: <400>28);
(m) LQRCFLYCSLFPKGHR (SEQ ID NO: <400>29);
(n) ELVHLWVAEG (SEQ ID NO: <400>30);
(o) WVAEG (SEQ ID NO: <400>31);
(p) NELVHLWVAEG (SEQ ID NO: <400>32); and (q) ELVHLWVAEGF (SEQ ID NO: <400>33) or a homologue, analogue or derivative thereof.
The present invention extends to a recombinant gene product that contains the above amino acid sequence motifs in any relative combination or frequency.
Preferably the recombinant gene product is capable of conferring, enhancing or facilitating pathogen resistance in a cell.
The recombinant pathogen resistance gene product, pathogen resistance-like gene product, or functional derivative thereof, may be used to produce immunoiogically 'interactive molecules, such 'as antibodies, or functional derivatives thereof, the only requirement being that the recombinant products ace irnmunologically interactive with antibodies to all or part of said gene product.
According to this aspect, there is provided an antibody which is capable of binding to a Rp1-D variant polypeptide wherein said polypeptide comprises substantially the same as the amino acid sequence set forth in SEQ ID NO: <400>2 andlor SEQ ID
NO:
<400>4 and/or SEQ ID NO: <400>6 and/or SEQ ID NO: <400>8 or is at least about 60% similar to all or at least about 5 contiguous amino acids thereof or an immunologically or functionally equivalent conformational epitope thereof.
Antibodies to a recombinant pathogen resistance gene product are particularly useful in the screening of plants for the presence of said gene product.
Accordingly, antibodies which are capable of binding to the primary amino acid sequence of the pathogen-resistance polypeptide described herein (i.e. a linear epitope) andlor to an epitope thereof which comprises the secondary, tertiary or quaternary structure of said polypeptide (i.e. a conformational epitope) are useful for such appiications. Those skilled in the art will be aware that an antibody preparation which is capable of recognising a specific polypeptide may comprise a population of molecules which recognise collectively linear and conformational epitopes.
Antibodies may be monoclonal or polyclonal and may be selected from naturally occurring antibodies to a pathogen resistance gene product or may be specifically raised to a recombinant or synthetic pathogen resistance gene product.
The pathogen resistance gene product may first need to be associated with a carrier molecule if it is insufficiently immunogenic without such an association.
Alternatively, fragments of antibodies may be used such as Fab fragments.
Furthermore, the present invention extends to recombinant and synthetic antibodies and to antibody hybrids. A "synthetic antibody" is considered herein to include fragments and hybrids of antibodies. The antibodies andlor the recombinant pathogen resistance gene products of the present invention are particularly useful for the immunological screening of pathogen resistance gene products in various plants, in monitoring expression of pathogen resistance genetic sequences in transgenic plants and as a proprietary tagging system.
In one embodiment, specific antibodies are used to screen for pathogen resistance gene products or pathogen resistance-like gene products in plants. Techniques for the assays contemplated herein are known in the art and include, for example, sandwich assays and ELISA.

It is within the scope of this invention to include any second antibodies (monoclonal, polyclonal or fragments of antibodies) directed to the first mentioned antibodies discussed above. Both the first and second antibodies may be used in detection assays or a first antibody may be used with a commercially available anti-s immunoglobulin antibody. An antibody as contemplated herein includes any antibody specific to any region of a recombinant pathogen resistance gene product.
Both polycional and monoclonal antibodies are obtainable by immunisation with a recombinant pathogen resistance gene product and either type is utilisable for immunoassays. The methods of obtaining both types of sera are well known in the art.
Polyclonal sera are less preferred but are relatively easily prepared by injection of a suitable laboratory animal with an effective amount of recombinant pathogen resistance gene product, or antigenic or immunointeractive parts thereof, collecting serum from the animal and isolating specific sera by any of the known immunoadsorbent techniques. Although antibodies produced by this method are utilisable in virtually any type of immunoassay, they are generally less favoured because of the potential heterogeneity of the product.
The use of monoclonal antibodies in an immunoassay is particularly preferred because of the ability to produce them in large quantities and the homogeneity of the product.
The preparation of hybridoma cell lines for monoclonal antibody production derived by fusing an immortal cell line and lymphocytes sensitised against the immunogenic preparation can be done by techniques which are well known to those who are skilled in the art (see, for example, Douillard and Hoffman, 1981; Kohler and Milstein, 1975).
The presence of a pathogen resistance gene product or pathogen resistance-like gene product in a plant or more commonly a plant extract may be accomplished in a number of ways such as by Western blotting and ELISA procedures. A wide range of immunoassay techniques are available as can be seen by reference to US Patent NOS. 4,016,043, 4, 424,279 and 4,018,fi53. These, of course, include both single-site and two-site or "sandwich" assays of the non-competitive types, as well as in the traditional competitive binding assays. These assays also include direct binding of a labelled antibody to a target.
Sandwich assays are among the most useful and commonly used assays and are favoured for use in the present invention. A number of variations of the sandwich assay technique exist, and all are intended to be encompassed by the present invention. Briefly, in a typical forward assay, an unlabelled antibody is immobilised on a solid substrate and the sample to be tested brought into contact with the bound molecule. After a suitable period of incubation, for a period of time and under conditions sufficient to allow formation of an antibody-antigen complex, a second antibody specific to the antigen, labelled with a reporter molecule capable of producing a detectable signal is then added and incubated, allowing time sufFcient for the formation of another complex of antibody-antigen-labelled antibody. Any unreacted material is washed away, and the presence of the antigen is determined by observation of a signal produced by the reporter molecule.
In this case, the first antibody is raised to a recombinant pathogen resistance gene product and the antigen is a pathogen resistance gene product in a plant.
The results may either be qualitative, by simple observation of the visible signal, or may be quantitated by comparing with a control sample containing known amounts of hapten. Variations on the forward assay include a simultaneous assay, in which both sample and labelled antibody are added simultaneously to the bound antibody.
These techniques are well known to those skilled in the art, including any minor variations as will be readily apparent. In accordance with the present invention the sample is one which might contain pathogen resistance gene product and include crude or purified plant extract such as extracts of (eaves, roots and stems.
In the typical forward sandwich assay, a first antibody raised against a recombinant pathogen resistance gene product is either covalently or passively bound to a solid surface. The solid surface is typically glass or a polymer, the most commonly used polymers being cellulose, polyacrylamide, nylon, polystyrene, polyvinyl chloride or polypropylene. The solid supports may be in the form of tubes, beads, discs of microplates, or any other surface suitable for conducting an immunoassay. The binding processes are well-known in the art and generally consist of cross-linking, covalent binding or physically adsorption, the polymer-antibody complex is washed in preparation for the test sample. An aliquot of the sample to be tested is then added to the solid phase complex and incubated for a period of time sufficient (e.g.

minutes) and under suitable conditions (e.g. 25°C) to allow binding of any antigen present in the sample to the antibody. Following the incubation period, the reaction locus is washed and dried and incubated with a second antibody specific for a portion of the first antibody. The second antibody is linked to a reporter molecule which is used to indicate the binding of the second antibody to the hapten.
An alternative method involves immobilising the target molecules in the biological sample and then exposing the immobilised target to specific antibody which may or may not be labelled with a reporter molecule. Depending on the amount of target and the strength of the reporter molecule signal, a bound target may be detected by direct labelling with the antibody. Alternatively, a second labelied antibody, specific to the first antibody is exposed to the target-first antibody complex to form a target-first antibody-second antibody tertiary complex. The complex is detected by the signal emitted by the reporter molecule.
By "reporter molecule" as used in the present specification, is meant a molecule which, by its chemical nature, provides an analytically identifiable signal which allows the detection of antigen-bound antibody. Detection may be either qualitative or quantitative. The most commonly used reporter molecules in this type of assay are either enzymes, fluorophores or radionuclide containing molecules (i.e.
radioisotopes) and chemiluminescent molecules.
In the case of an enzyme immunoassay, an enzyme is conjugated to the second antibody, generally by means of glutaraldehyde or periodate. As will be readily recognised, however, a wide variety of different conjugation techniques exist, which are readily available to the skilled artisan. Commonly used enzymes include horseradish peroxidase, glucose oxidase, beta-galactosidase and alkaline phosphatase, amongst others. The substrates to be used with the speck enzymes are generally chosen for the production, upon hydrolysis by the corresponding enzyme, of a detectable colour change. Examples of suitable enzymes include alkaline phosphatase and peroxidase.
It is also possible to employ fluorogenic substrates which yield a fluorescent product rather than the chromogenic substrates noted above. In all cases, the enzyme-labelled antibody is added to the first antibody-hapten complex, allowed to bind, and then the excess reagent is washed away. A solution containing the appropriate substrate is then added to the complex of antibody-antigen-antibody. The substrate will react with the enzyme linked to the second antibody, giving a qualitative visual signal, which may be further quantitated, usually spectrophotometrically, to give an indication of the amount of hapten which was present in the sample. The term "reporter molecule"
also extends to use of cell agglutination or inhibition of agglutination such as red blood cells on latex beads, and the like.
Alternately, fluorescent compounds, such as fluorescein and rhodamine, may be chemically coupled to antibodies without altering their binding capacity. When w activated by illumination with light of a particular wavelength, the fluorochrome-labelled antibody adsorbs the light energy, inducing a state to excitability in the molecule, followed by emission of the light at a characteristic colour visually detectable with a light microscope. As in enzyme immunoassays (EIA), the fluorescent labelled antibody is allowed to bind to the first antibody-hapten complex. After washing off the unbound reagent, the remaining tertiary complex is then exposed to the light of the appropriate wavelength the fluorescence observed indicates the presence of the hapten of interest.
Immunofluorescene and EIA techniques are both very well established in the art and are particularly preferred for the present method. However, other reporter molecules, such as radioisotope, chemiluminescent or bioluminescent molecules, may also be employed.

It will be readily apparent to the skilled technician how to vary the above assays and all such variations are encompassed by the present invention.
In an alternative embodiment, the hybridisation and PCR techniques described supra may be utilised with any necessary modifications to determine the presence of a pathogen-resistance gene in a plant or to quantitate the level of expression of said gene at the RNA level.
Those skilled in the art will be aware that the genetic construct described supra may be used to "transfect" a cell, in which case it is introduced into said cell without integration into the cell's genome. Alternatively, a genetic construct may be used to "transform" a cell, in which case it is stably integrated into the genome of said cell.
Accordingly, the isolated nucleotide sequence of the present invention, or homologue, analogue or derivative thereof, may be introduced into a cell using any known method for the transfection or transformation of said cell, thereby conferring enhanced pathogen-resistance properties thereon. Wherein a cell is transformed by the genetic construct of the invention, a whole organism may be regenerated from a single transformed cell, using any method known to those skilled in the art.
Accordingly, a further aspect of the invention extends to a plant such as a crop plant carrying a non-endogenous Rp9-D variant or Rpg1 variant which encodes a poiypeptide which confers, enhances, or otherwise facilitates pathogen resistance in said plant. Preferably, the plant is a monocot plant. More preferably the transgenic plant is one or more of the following: wheat, maize, barley, rye, oats, pearl millet, rice, sorghum, sugar cane, Tiiticum tauschii or ryegrass, amongst others. Other species are not excluded.
In fact, the non-endogenous Rp9-D variant genetic sequence, Rpg1 variant genetic sequence or transgene may originate from any plant species. Preferably, said genetic sequence is identical to any one or more of the nucleotide sequences set forth in SEQ

ID NOS: <400>1 or <400>3 or <400>5 or <400>7, or <400>64 to <400>70 or a functional derivative, fragment, part, complement, homologue, or analogue thereof.
Furthermore, wherein said genetic sequence or transgene is a cDNA molecule such as set forth in SEQ ID NO: <400>3 or other nucleic acid molecule which lacks a functional promoter, it may be placed operably under control of promoter sequence.
The expression of the transgene may be constitutive or inducible by an external stimulus such as physiological stress, or by addition of a chemical compound, or the expression may be developmentally-regulated, or expressed in a tissue- or cell-specific pattern. Furthermore the transgene may be inserted into or fused to a particular endogenous genetic sequence. Methods for placing a structural gene operably under the control of a promoter sequence are welt-known to those skilled in the art.
A genetic construct designed to express a recombinant Rp1-D variant polypeptide or variant Rpg1 polypeptide may be introduced into plant tissue, thereby producing a "transgenic plant", by various techniques known to those skilled in the art.
The technique used for a given plant species or specific type of plant tissue depends on the known successful techniques. Means for introducing recombinant DNA into plant tissue include, but are not limited to, direct DNA uptake into protoplasts (Krens et al, 1982; Paszkowski et al, 1984), PEG-mediated uptake to protoplasts (Armstrong et al, 1990) microparticle bombardment electroporation (Fromm ef al., 1985), microinjection of DNA (Crossway et al., 1986), microparticle bombardment of tissue explants or cells (Christou et al, 1988; Sanford, 1988) or T-DNA-mediated transfer from Agrobacterium to the plant tissue. Methods for the Agrobacterium-mediated transformation of plants will be well-known to those skilled in the art. In particular, methods for the Agrobacferium-mediated transformation of rice (Oryza satlva) tissue have been disclosed by Heie et aL. Representative T-DNA vector systems are described in the following references: An et aG(1985); Herrera-Estrella et al. (1983a,b);
Herrera-Estrella ef al. (1985).
For microparticle bombardment of cells, a microparticle is propelled into a plant cell, _4q._ in particular a plant cell not amenable to Agrobacterium mediated transformation, to produce a transformed cell. Wherein the cell is a plant cell, a whole plant may be regenerated from the transformed plant cell. Alternatively, other non-animal cells derived from multicellular species may be regenerated into whole organisms by means known to those skilled in the art. Any suitable ballistic cell transformation methodology and apparatus can be used in practising the present invention. Exemplary apparatus and procedures are disclosed by Stomp et al. (U.S. Patent No. 5,122,4fifi) and Sanford and Wolf (U.S. Patent No. 4,945,050). When using ballistic transfom~ation procedures, the genetic construct may incorporate a plasmid capable of replicating in the cell to be transformed.
Examples of microparticles suitable for use in such systems include 1 to 5 ~cm gold spheres. The DNA construct may be deposited on the microparticle by any suitable technique, such as by precipitation.
Plant species may be transformed with the genetic construct of the present invention by the DNA-mediated transformation of plant cell protoplasts and subsequent regeneration of the plant from the transformed protoplasts in accordance with procedures well known in the art.
Any plant tissue capable of subsequent clonal propagation, whether by organogenesis or embryogenesis, may be transformed with a vector of the present invention.
The particular tissue chosen will vary depending on the clonal propagation systems available for, and best suited to, the particular species being transformed.
Exemplary tissue targets include leaf disks, pollen, embryos, cotyledons, hypocotyls, megagametophytes, callus tissue, existing meristematic tissue (e.g., apical meristem, axillary buds, and root meristems), and induced meristem tissue (e.g., cotyledon meristem and hypocotyl meristem).
The term "organogenesis", as used herein, means a process by which shoots and roots are developed sequentially from meristematic centres.

The term "embryogenesis", as used herein, means a process by which shoots and roots develop together in a concerted fashion (not sequentially), whether from somatic cells or gametes.
Plants of the present invention may take a variety of forms. The plants may be chimeras of transformed cells and non-transformed cells; the plants may be clonal transformants (e.g., all cells transformed to contain the expression cassette); the plants may comprise grafts of transformed and untransformed tissues (e.g., a transformed root stock grafted to an untransformed scion in citrus species). The transformed plants may be propagated by a variety of means, such as by clonal propagation or classical breeding techniques. For example, a first generation (or T1 ) transformed plants may be selfed to give homozygous second generation (or T2) transformed plants, and the T2 plants further propagated through classical breeding techniques.
The genetic construct may further incorporate a dominant selectable marker, such as npfll, hygromycin-resistance gene, a phosphinothricin-resistance gene or ampiciliin-resistance gene, amongst others, associated with the transforming DNA to assist in cell selection and breeding.
Once introduced into the plant tissue, the expression of the introduced gene may be assayed in a transient expression system, or it may be determined after selection for stable integration within the plant genome. Techniques are known for the in vitro culture of plant tissue, and in a number of cases, for regeneration into whole plants.
Procedures for transferring the introduced genetic construct from the originally transformed plant into commercially useful cultivars are known to those skilled in the art.
In the context of the present invention, it is preferred that the transgenic plants thus generated exhibit enhanced pathogen-resistance properties compared to otherwise isogenic non-transformed plants, in particular against fungal pathogens and viral pathogens. For example, Rp1-D andlor Rpg? and/or hrpl variant genes which confer or enhance rust-resistance can be introduced into cereals to produce novel lines with improved resistance to rusts, as an alternative or adjunct to conventional plant breeding approaches. In particular, Rp1-D andlor other Rp alleles may be used to confer resistance against rusts such as Puccinia ssp. in maize, sweet corn, wheat and sorghum, amongst others. Similarly, the barley Rpg1 variant alleles exemplified herein may be used to confer resistance against barley stem rust, amongst others.
Whilst rice does not have a rust pathogen, it does contain Rpl-D variant genes which may be useful in conferring resistance against rice pathogens such as the rice blast Pyricularia oryzae or the rice blight Xanfhomonas oryzae, amongst others. Similarly, the Rp1-D
variant of wheat may be used to confer resistance against wheat streak mosaic virus, amongst others in cereals.
The present invention extends to the progeny and clonal derivatives of said transgenic plant.
The present invention is further described in the following Examples. The embodiments exemplified hereinafter are in no way to be taken as limiting the subject invention.

Strategy for isolating Rp1-D variant genes The Rp1-D rust resistance gene was isolated from maize by transposon tagging using two independent maize transposon systems (Mu and AclDs). The two methods resulted in the isolation of the same identical DNA sequence: the Rp1-D gene, which belongs to the NBS-LRR class of plant resistance gene.
The Rp1-D rust resistance gene was isolated from maize after tagging with the maize transposon Mu. A DNA fragment, identified with a Mu probe, is absent in the rust resistant parent and present in the susceptible transposon-tagged mutant. This new fragment co-segregates with the mutant Rp1-D allele. DNA flanking the Mu insertion in the novel fragment was cloned and sequenced. The putative translation product of the flanking DNA sequence was shown to encode a leucine-rich repeat nucleotide binding site polypeptide (SEQ 1D NO:<400>2 or SEQ ID NO: <400>4).
In a further approach, the Rp1-D rust resistance gene was isolated from maize after tagging with the maize transposon Ds. A mutant allele was identified and cloned using a DNA probe encoding a resistance gene analogue sequence derived by PCR using pools of degenerate primers designed to encode conserved amino acid sequence motifs present in NBS-LRR resistance genes (Table 2). The Ds element is absent in the wild-type allele, appears in the mutant allele and is absent in rust resistant revertants of the Ds allele.
The Ds containing allele has the identical sequence of the gene tagged by the Mu transposon. Both strategies used to isolate the Zea mays Rp1-D allele set forth in SEQ ID NO: <400>1 thus relied upon the transposon mutagenesis of Zea mat's and required detailed genetic data to facilitate linkage analysis demonstrating co-segregation of the resistance phenotype or susceptible phenotype with the wild type allele or mutant allele, respectively. In this regard, absent any linkage data mere amplification approaches alone would have presented considerable difficulties in assigning a function to the ampl~ed NBS-LRR encoding nucleotide sequences obtained, particularly in consideration of the diversity of such sequences present in plant genomes. Notwithstanding that this is the case, those skilled in the art will recognise that once the function of the nucleotide sequence of the Zea mat's Rp1-D
allele set forth in SEQ ID NOS: <400>1 and <400>3 was established by the present inventors, said sequence provided a means for isolating andlor detecting functionally-equivalent sequences in Zea mat's or other plants.

Isolation of Rp1-D sequences using the Mutator transposable element system Families which were heterozygous for two different Rp1 alleles (Rp1-DlRp1-A
and Rp1-DlRp1-B) with multiple, active Mutator transposable elements were constructed and out crossed to Rp1-J homozygotes. The resulting families were screened with the common rust (P.sorghr) biotype IN2, which is virulent on lines carrying Rp1-J, but avirulent on lines carrying Rp1-D, Rp1-A and Rp1-B. After testing 150,000 maize seedlings with biotype IN2, 55 susceptible individuals were identified. DNA
from the susceptible individuals were analysed with the RFLP probes umc285 and bn13.04 which detect RFLP loci which flank the locus. This permitted the distinction between susceptible individuals which arose from cross over events from those which did not, the tatter being candidates for transposon insertion mutants. Four of the susceptible individuals had flanking RFLP marker alleles of the Rp1-D parent, indicating they were derived from mutations of the Rp1-D allele and were not derived from crossing-over.
Each of the four mutant individuals was crossed two or three times to maize lines which carried detectable rp1 alleles but no Mutator elements to reduce the number of Mutator elements in the line. DNAs of families derived from each of the four mutants, segregating for the mutant allele, were then analysed by gel blot analysis with several Mutator DNA probes to determine if any of the lines carried Mutator elements which mapped to the rp1 locus. No mutator elements which mapped to the rp1 locus were identified in three of the four families.
In the family segregating for the fourth mutation, a Mutator probe hybridized to a Hindlll fragment of approximately 5 Kb which cosegregated with the mutant allele in 90 progeny. After self fertilizing an individual carrying the mutant allele, an individual which was homozygous for the mutant allele was identified, DNA was purified and a library of 4-6 kb Hindlll fragments was made in a Lambda cloning vector. The 5 Kb Hindlll fragment which mapped to rp1 was selected using a Mutator probe and the 5 Kb Hindlll fragment was sequenced. The clone was found to carry a Mu2 element inserted into approximately 3 Kb of DNA with sequence homology to known resistance genes.
VlJhen used as a probe in gel-blot analysis, the sequences flanking the Mu2 element detect a gene family which maps to the rp1 locus.

OLIGONUCLEOTIDE PRIMERS USED TO AMPLIFY Rp1 ALLELES
Primer Primer sequence (5' to 3') SEG1 ID NO:

p-IoopAA* AAG AAT TCG GNG TNG GNA AAA CAA C <400>34 p-IoopAT* AAG AAT TCG GNG TNG GNA AAA CTA C <400>35 p-IoopAC* AAG AAT TCG GNG TNG GNA AAA CCA C <400>36 p-IoopAG' AAG AAT TCG GNG TNG GNA AAA CGA C <400>37 p-IoopGA* AAG AAT TCG GNG TNG GNA AGA CAA C <400>38 p-IoopGT* AAG AAT TCG GNG TNG GNA AGA CTA C <400>39 p-IoopGC* AAG AAT TCG GNG TNG GNA AGA CCA C <400>40 p-IoopGG* AAG AAT TCG GNG TNG GNA AGA CGA C <400>41 kinase-2D CTA CTG NTN CTN GAC GAC GT <400>42 kinase-2E CTA CTG NTN CTN GAC GAT GT <400>43 kinase-2F CTA CTG NTN CTN GAT GAC GT <400>44 kinase-2G CTA CTG NTN CTN GAT GAT GT <400>45 GLPL1* AAC TCG AGA GNG CNA GNG GNA GGC C <400>46 GLPL2* AAC TCG AGA GNG CNA GNG GNA GAC C <400>47 GLPL3* AAC TCG AGA GNG CNA GNG GNA GTC C <400>48 GLPL4* AAC TCG AGA GNG CNA GNG GNA GCC C <400>49 GLPLS* AAC TCG AGA ANG CCA ANG GCA ATC C <400>50 GLPL6* AAC TCG AGA ANG CCA ANG GCA AAC C <400>51 CFA1 CA(AlG} (TIA)AI GC(GIA) AA(GIA) CA(CIT)<400>52 TGT TT

CFA2 CA(AIG) (TIA)AI GC(GIA) AA(GIA) CA(C!T)<400>53 TGC TT

CFA3 ATA GA(AIG) CA(AIG) (T/A)AI GC(G/A) <400>54 AAA CA

CFA4 ATA GA(AIG) CA(A!G) (TIA)AI GC(GIA) <400>55 AAG CA

WMA1 A(TIC)(A/G) AAN CCN TNA GCC ATC CA <400>56 VUMA2 A(TIC)(AIG) AAN CCN TNT GCC ATC CA <400>57 WMA3 A(T/C)(AIG) AAN CCN TNC GCC ATC CA <400>58 WMA4 A(TIC)(AIG} AAN CCN TNG GCC ATC CA <400>59 MHD1 I CGA CAG TCN ATC ATG CAT I <400>60 MHD2 I CGA CAG TCN ATC GTG CAT I <400>61 MHD3 I CGA CAG TCN GTC ATG CAT I <400>62 I MHD4 I CGA CAG TCN GTC GTG CAT ~ <400>63 ' Primers are the same as those described by R. Michelmore (University of California, Davis) except foi the 5'ends. Primer names accompanied by an asterisk to indicate these differences.

Protocol for the isolation of Rp1-D sequences using the Ds transposable element system Plants that were homozygous for the Rp1-D13 allele and heterozygous for the Pw Ac containing gene were crossed by plants containing the Rp5 rust resistance gene which is linked and about 1-2 cm distal on the short arm of chromosome 10 (Figure 1). A
number of susceptible mutants were recovered from 10,000 progeny screened.
These were selfed and used to isolate homozygous mutants of the Rp1-D13 allele.
Several of these were tested for reversion to resistance in the presence of Ac activity. The susceptible mutant rp1-D13-2 reverted to resistance in 212800 test cross progeny providing genetic evidence for tagging with a Ds transposon. The PIC20 DNA
clone, containing a resistance gene analogue sequence, reveals a small gene family that in Southern analysis co-segregates with the rp1 locus (Figures 2A and 2B). When used for analysis of the rp1-13-2 mutant and resistant revenants, the PCI20 probe demonstrated the presence of an approximately 400 by insertion in rp1-D-13-2 which was absent in the resistant revertants (Figure 2A, compare lanes 22 and 23).
Cloning and sequencing of the band with the altered size confirmed the presence of the Ds insertion and the flanking DNA sequence was shown to be identical to the sequences isolated independently by the Mu transposon tagging experiment.

Rp1-D DNA Probes can detect and clone resistance genes in other cereals A DNA probe isolated from the nucleotide binding site region of the rp1 gene detected RFLPs between barley varieties Steptoe and Morex (Figure 3). Morex contains the WO 99/45118 PCTlAU99/00130 Rpg1 rust resistance gene and is resistant to barley stem rust, while Steptoe lacks this gene and is susceptible. A mapping family derived from doubled haploids of a Steptoe X Morex F1 plant (Ref) which has been scored for Rpg1 resistance was used to map the rp1 RFLPs. One RFLP co-segregated for Rpg1 in 150 progeny (Figure 3). Two other unlinked loci were also detected with the Rp1 probe which may correspond to other cereal disease resistance loci. Two alleles of the gene family at the Rpg1 locus were cloned and sequenced, designated Rpg1-2 (SEQ ID NO:<400>5) and Rpg1-13 (SEQ ID NO:<400>7). ClustalW analyses indicate significant homologies between the Rp1-D, Rpg1-2 and Rpg1-13 alleles, at both the nucleotide level and the amino acid level (Figures 4A and 5). The Rpg1-2 allele is 58% identical to Rp1-D at the protein level, whilst the Rpg1-13 allele is 60% identical to Rp1-D (Figure 4A; Figure 5). The Rpg1-2 and Rpg1-13 proteins are 74% identical. Accordingly, the term "at least about 60% identical" shall be understood to include 57% or 58% or 59% or 60%
identity.

Identification of Rp1-D variants in other plant species Genomic DNA was isolated from seedlings of sorghum, Panicum ssp., maize, sugar cane, barley, hexaploid wheat, oats, Triticum tauschii and rice, digested with either Ncol or Bglli and subjected to Southern hybridisation using a maize Rp1-D
nucleotide sequence as a probe. As shown in Figure 6, hybridising bands were present in all species tested, suggesting that Rp1-D variant alleles are present in a wide range of different monocotyledonous plants.

Coding regions of Rp1-D homologous genes Rp1-D homologous genes were cloned from Zea mays and the coding regions determined. The nucleotide sequences of these coding regions are shown in SEQ
ID
NOS:<400> 64 to <400> 70. The homologous genes are referred to herein as hrp1-d1 to hrp1-d6 (SEQ ID NOS:<400>64 to <400>69, respectively and hrp1-cin4 (SEQ
ID
NO:<400>70).

REFERENCES
1. An et al. (1985) EMBO J 4:277-284.
2. Armstrong, et al.Planf Cell Reports 9: 335-339, 1990.
3. Ausubel, et al (1992}, Current Protocols in Molecular Biology, Greene~ley, New York.
4. Christou, P., et al. Plant Physiol 87: 671-674, 1988.
5. Crossway et aL, Mol. Gen. Genet. 202:179-185, 1986.
6. Devereux, J., et al. {1984). Nucl. Acids Res. x:387-395.

7. Dixon, R., and Lamb, C (1990) Ann. Rev. Plant Physiol. Plant Mol. Biol.
41:339-367.

8. DouiUard and Hoffman (1981) !n: Compendium of Immunology Vol II (ed.
Schwarz).

9. Fromm ef al. Proc. Nafl. Acad. Sci. (USA) 82:5824-5828, 1985.

10. Herrera-Estella et al., Nature 303: 209-213, 1983a.

11. Herrera-Estella et aL,EMBO J. 2: 987-995, 1983b.

12. Herrera-Estella et al. In: Plant Genetic Engineering, Cambridge Universit)r Press, N.Y., pp 63-93, 1985.

13. Kohler and Milstein (1975) Nature 256: 495-499.

14. Krens, F.A., et al., Nature 296: 72-74, 1982.

15. Marineau, C., Matton, D.P. and Brisson, N. (1987) Plant Mol. Biol. 9: 335-342.

16. McPherson, M.J., Quirke, P. and Taylor, G.R. {1991} PCR A Practical Approach. IRL Press, Oxford University Press, Oxford, United Kingdom.

17. Needleman and Wunsch (1970) J. Mot. Biol. x$:443-453.

18. Paszkowski et al., EMBO J. 3:2717-2722, 1984 19. Sambrook et al (1989), Molecular Cloning: A Laboratory Manual, Cold Springs Harbor Laboratory Press 20. Sanford, J.C., et al., Particulate Science and Technology 5: 27-37, 1987.

21. Thompson,et al., (1994} Nucl. Acids Res. 22:4673-4680.

SEQUENCE LISTING
<110> COMMONWEALTH SCIENTIFIC AND INDUSTRIAL RESEARCH ORGANISATION
GRAINS RESEARCH AND DEVELOPMENT CORPORATION
KANSAS STATE UNIVERSITY RESEARCH FOUNDATION
<120> GENETIC SEQUENCES CONFERRING PATHOGEN RESISTANCE IN
PLANTS AND USES THEREFOR
<130> p:\oper\mro\rpl-d. pct <140>
<141>
<150> US 60/077109 <151> 1998-03-06 <160> 70 <170> PatentIn Ver. 2.0 <210> 1 <211> 5948 <212> DNA
<213> Zea mays <220>
<221> CDS
<222> i1200)..i5075) <400> 1 gtcgaccacc aacagacaat attttccctt tccacccact aactcttttc ttaaatcttt 60 ttatgatatc attataatct tcattatact aagctttctg tgatgcattg gaatacccag 120 atatttgaaa ggataatgcc ctttattaca taaaaaaatt ctgagtattc caattctcgt 180 tctttagtag catcataaca gaaaagttgg ctcttatgaa aatttatttt gagaccggaa 240 agatatacaa acgcacaaag tagtaagcca aaactgcatt caaaggaaac gtggaggctt 300 gtttccacac agccactatt acacggaaca caactgtaat catccagtgc cttgtttttc 360 ctcatcagaa ggttcctagt attgagtcta tctctaggat tctagaaaag tggaaccaaa 420 taggaccgaa agcgcagagg aactgctatg ccttaaggac tggatgcaaa ttggaaatga 480 aagagatagt tttttttagg attttggtct gtactggaag ctctcaattt ttatggccag 540 i WO 99/45118 PG"T/AU99/00130 atgaattgtt ggtccaactc atggggatct cgtcttctac tttcatatca agctcagctc 600 ctcttgcaac gtacaaccat gtctaaaaaa aataggggag gactccaagg aaagctccaa 660 aagctttctt tgtctgtgtt tattcactcg aactgcataa ttcagagcag tcccaatcga 720 tacactagct cccaacaata attcagagca gtcctaatcc atagactagt ctactccgct 780 tgttcaccgc ttgttcacct ttctctttcc atgctgcttc ctttcccagc gagcagtaga 840 ccctcagcag tctacggcac cttcggtctc ttctacataa aaaacaaatc atctatcacc 900 agcaatcagc aacaccatcc ggtattcctc cgttccctgc ccgacggsgc tgettccctc 960 cttgctgtgc tgtgcttgtt ccagattcta ctccatacgc ccttgattta ttttacttcg 10x0 tgtaagatct ccaatctcca cttttttcgc ttgctgtgtt agctactatt tcccctttgt 1080 gttgctggat ctggcgagaa agaaaaataa ttggtttgtg cttcattttt tcattcagat 1140 tcattaattt tatgtgtctg cagagaaaaa agaaaaaaaa aagcttctta ctgaatttc 1199 atg gcc gac ttg gcg ctc gcc ggc tta agg tgg gca gca tcg ccg att is47 Met Ala Asp Leu Ala Leu Ala Gly Leu Arg Trp Ala Ala Ser Pro Ile gtc aac gag ctt ctt act aaa get tca get tac ctc agt gtg gac atg 1295 Val Asn Glu Leu Leu Thr Lys Ala Ser Ala Tyr Leu Ser Val Asp Mat gtg cgt gag atc caa cga cts gaa gcc act gtc ctg cca cag ttc gag 1343 Val Arg Glu Ile Gln Arg Leu Glu Ala Thr Val Leu Pro Gln Phe Glu ctg gtg att caa gcg gcc cag aag agc ccc cac agg ggc ata ctg gag 1391 Leu Val Ile Gln Ala Ala G1n Lys Ser Pro His Arg Gly Ile Leu Glu gca tgg ctc cgg cgt ctc aaa gaa gcc tac tat gat gcc gag gac ttg 1439 Ala Trp Leu Arg Arg Leu Lye Glu Ala Tyr Tyr Asp Ala Glu Asp Leu 65 ?0 75 BO
ttg gac gag cat gag tac aat gtc ctt gag ggc aag gcc aag agc gaa 1487 Leu Asp Glu His Glu Tyr Asn Val Leu Glu Gly Lys Ala Lys Ser Glu aaa agt ctc ctg ctg gga gag cat gga agc tcc tcc act gca act act 1535 Lys Ser Leu Leu Leu Gly Glu Hia Gly Ser Ser Ser Thr Ala Thr Thr gtc atg aag cet ttt cat get get atg agc agg gca cgg aac ttg etc 1583 Val Met Lys Pro Phe His Ala Ala Met Ser Arg Ala Arg Asa Leu Leu 115 120 1~5 cct caa aac aga agg cta att agc aag atg aac gag ctc aaa gca atc 1631 Pro Gln Asn Arg Arg Leu Ile Ser Lys Met Asn Glu Lau Lys Ala Ile ctg aca gaa gcc caa caa ctt cga gat ctt ctt ggt ttg cca cat ggc 1679 Leu Thr Glu Ala Gln Gln Leu Arg Asp Leu Leu Gly Lau Pro His Gly aat acc gtc gag tgg cca get gca gca cct acc agt gtt ccc aca acc 1727 Asn Thr Val Glu Trp Pro Ala Ala Ala Pro Thr Ser Val Pro Thr Thr aca tca ctt ccc act tcc aag gtt ttt ggt cgc gac agg gat cgt gat 1775 Thr Ser Leu Pro Thr Bar Lys Val Phe Gly Arg Asp Arg Asp Arg Asp cgt ata gta gat ttt ctt ctc ggc aag aca aca act get gag gca agc 1823 Arg Ile Val Asp Phe Lsu Leu Gly Lys Thr Thr Thr Ala Glu Ala Ser tea get aag tac teg ggt ttg gcc att gtt gga ttg gga gga atg ggg 1871 Ser Ala Lys Tyr Ser Gly Leu Ala Its Val Gly Leu Gly Gly Met Gly 210 215 Za0 aag tcc acc tta gca cag tat gtc tat aat gac aaa agg ata gaa gaa 1919 Lys Ser Thr Leu Ala Gln Tyr Val Tyr Asa Asp Lys Arg Ile Glu Glu Za5 230 a35 240 tgc ttt gat atc agg atg tgg gtg tgc atc tca cgc saa ctt gat gtg 1967 Cys Phe Aap Ile Arg Met Trp Val Cys Ile Ser Arg Lye Leu Asp Val cat cgt cac aca agg gag att ata gag tct gca aaa asg gga gag tgc 2015 His Arg His Thr Arg Glu Ile Ile Glu Ser A1a Lys Lys Gly Glu Cys cca cgt gtt gat aat ctc gat act ctc cag tgc aaa tta cgt gat ata 2063 Pro Arg Val Asp Asn Leu Asp Thr Leu Gln Cys Lys Leu Arg Asp Ile cta caa gag tca cag aea ttc ctg ctt gtc ttg gat gat gtt tgg ttt Gill Leu Gln Glu Ser Gln Lys Phe Leu Leu Val Leu Asp Asp Val Trp Bhe a90 295 300 gaa aaa tct cat aat gag aca gag tgg gag tta ttc ctt get cca tta 2159 Glu Lys Ssr His Asn Glu Thr Glu Trp Glu Leu Fha Leu Ala Pro Leu gtc tct aaa cag tca ggg agc aaa gtt ttg gtg act tct cga agt saa ZZ07 Val Sex Lys Gln Ssr Gly Ser Lys Val Leu Val Thr Ser Arg Ser Lys aca ctt cct gcc get att tgt tgt gaa caa gaa cat gtc att cat ttg 2x55 Thr Leu Pro Ala Ala Ile Cys Cys Glu Gln Glu His Val Ile His Leu aaa aac atg gat gat act gag ttt ttg get ctt ttt aaa cac cat get 2303 Lys Asn Met Asp Asp Thr Glu Phe Leu Ala Leu Phe Lys His His Ala ttc tct gga gca gas atc aaa gac caa gta tta cgc acg aag ctg gaa x351 Phe Ser G1y Ala Glu Ile Lys Asp Gln Val Leu Arg Thr Lys Leu Glu gac act gca gtg gag stt get aaa agg ctt gga caa tgt cct ttg gca 2399 Asp Thr Ala Val Glu Ile Ala Lys Arg Leu Gly Gln Cars Pro Leu Ala gca aaa gtt ctg ggt tct cga ttg tgc agg aaa aag gat att get gaa 2447 Ala Lys Val Lau Gly Ser Arg Lau Cys Arg Lys Lys Asp Ile Ala Glu tgg aaa get get cta aag att gga gat tta agt gat ccc ttc aca tct 2495 Trp Lys Ala Ala Leu Lys Ile Gly Asp Leu Ser Asp pro Phe Thr Ser ctg ttg tgg agt tac gag aag tta gat cca cgt ctg cag agg tgc ttc 2543 Leu Leu Trp Ser Tyr Glu Lys Leu Asp 8ro Arg Leu Gln Arg Cys She ttg tat tgc agc ttg ttt cca aaa ggt cat aga tat gaa tct aat gag 2591 Lau Tyr Cys Ser Leu Phe Pro Lys Gly His Arg Tyr Glu Ser Asn Glu ttg gtt cac ctt tgg gtg gca gaa gga ttt gtt ggt tca tgc aat ttg 2639 Leu Val His Leu Trp Val Ala Glu Gly Phe Val Gly Ser Cys Asn Leu agt agg aga acg tta gaa gag gtt ggg atg gat tac ttc aat gat atg 2687 Ser Arg Arg Thr Lsu Glu Glu Val Gly flat Aep Tyr Phs Asn Asp list gtc tct gta tct ttc ttc caa ttg gtt ttt cat atc tat tgt gat tcg 2735 Val Ser Val 8er Pha Phs Cln Leu Val Phe His Ile Tyr Cys Asp Ser tac tat gtc atg cat gat atc ctt cat gat ttt gca gag tca ctc tct 2783 Tyr Tyr Val Met His Asp Ile Lsu His Asp Phe Ala Glu Ser Leu Ser 515 5a0 525 aga gaa gac tgc ttt aga tta gaa gat gat aat gtg aca gaa ata cca x831 Arg Glu Asp Cys Phs Arg Leu Glu Asp Asp Asn Val Thr Glu Its Pro tgc act gtt cga cat cta tct att cat gtt cat agt atg caa aag cat 2879 Cye Thr Val Arg His Leu Ser Ile Hie Val His Ssr Met Gln Lys His aag caa att atc tgc aag cta cat cat tta cgc act att atc tgc atc 2937 Lys Gln Zle Ile Cya Lys Leu His His Leu Arg Thr Ile Ile Cys Its gat ccg cta atg gat ggc cca agt gat att ttt gat ggc atg cta cgg 2975 Asp Pro Lsu Met Asp Gly Pro Ser Asp Ile Phe Asp Gly Met Lsu Arg aac caa aga aaa ctg cgt gta ttg tct ctg tca ttt tac aac agc aaa 3023 Asn Gln Arg Lys Leu Arg Val Leu Ser Leu Ser Phs Tyr Asn Ser Lys aat ttg cca gas tct att ggt gag ctg aag cac ctc cgg tat ttg aac 3071 Asn Lsu Pro Glu Ser Its Gly Glu Lau Lys His Lau Arg Tyr Leu Asn ctc atc agg acg tta gtt tct gaa ttg cct aga tca tta tgt act ctc 3119 Leu Ile Arg Thr Lsu Val Ser Glu Leu Pro Arg Ser Leu Cys Thr Lsu 6a5 630 635 640 tac cac tta caa tta ctt tgg tta aac cac atg gtg gag aat ttg cct 3167 Tyr His Leu Gln Leu Leu Trp Lsu Asn His Met Val Glu Asn Leu Pro gac aaa cta tgc aat tta aga aag cta cga cat cta gga gcg tac gtg 3215 Asp Lys Leu Cys Asn Leu Arg Lye Leu Arg His Lsu Gly Ala Tyr Val aat gat ttc gcg att gaa aag act att tgc caa att ctg aat ata ggt 3263 Asn Asp Phe Ala Ile Glu Lye Pro Ile Cys Gln Ile Leu Asn Ila Gly aag tta acg tcg cta caa cac att tat gtc ttt tct gta caa aag aag 3311 Lys Leu Thr Ser Leu Gln His Ile Tyr Val Phe Ser Val Gln Lys Lys caa ggc tat gag ttg cga cag ttg aag gac ttg aat gag ctt ggt ggc 3359 Gln Gly Tyr Glu Leu Arg Gln Leu Lys Asp Leu Asn Glu Leu Gly Gly agt tta aaa gtg aaa aat ctt gag aat gtc att gga aag gat gaa gcc 3407 Ser Leu Lys Val Lys Asn Leu Glu Asn Val Ile Gly Lys Asp Glu Ala gta gag tcg aag cta tat ctg aaa agt cgc ctt aaa gag ttg gca ctt 3455 Val Glu Ser Lys Leu Tyr Leu Lys Ser Arg Leu Lys Glu Leu Ala Leu gag tgg agt tcc gag aat gga atg gat gca atg gat att cta gaa ggt 3503 Glu Trg Ser Ser Glu Asa Gly Met Asp Ala Met Asp Ile Leu Glu Gly ctg aga cca cea ccc caa ctg agt aag ctc aca atc gaa ggt tac aga 3551 Leu Arg Pro Pro Pro Gln Leu Ser Lys Lsu Thr Ile Glu Gly Tyr Arg tct gat aca tat cct ggg tgg tta cta gag cga tcc tat ttt gag aat 3599 Ser Asp Thr Tyr Pro Gly Trp Leu Leu Glu Arg Ser Tyr Bhe Glu Asa ttg gaa agt ttt cag ctt agt aat tgc agt ttg cta gaa ggc cta cca 3647 Leu Glu Ser Phe Gln Lau Ser Asn Cys Ser Leu Leu Glu Gly Leu Pro cca gat aca gag ctc ctt cgg aat tgc tct agg ttg cgt sta aac ttt 3695 Pro Asp Thr Glu Leu Leu Arg Asn Cys Ser Arg Leu Arg Ile Asn Phe gtt cca aat ttg aag gaa cta tct aat ctt cca gca ggc ctt aca gat 3743 Val Pro Asn Leu Lys Glu Leu Ser Asn Leu Pro Ala Gly Leu Thr Asp tta tca att ggt tgg tgc cca ctg ctt atg ttt atc acc aac aat gag 3791 Leu Ser Ile Gly Trp Gars Pro Lau Leu Met Phe Ila Thr Asn Asn Glu cta gga cag cat gac ttg agg gaa aat ata ata atg aag gca gcc gac 3839 Lsu Gly Gln His Asp Leu Arg Glu Asn Ile Ile Met Lys Ala Ala Asp ctg gca tct aaa ctt gca ttg atg tgg gag gtg gat tca gga aaa gaa 3887 Leu Ala Ser Lys Leu Ala Leu Met Trp Glu Val Asp Sar Gly Lye Glu gtt agg aga gta ctg ttt gaa gac tat gta tct ctg att cgg ttg atg 3935 Val Arg Arg Val Leu Phe Glu Asp Tyr Val Ser Leu Ile Arg Leu Met aca ttg atg atg gat gat gat ata tca aag cat ctt caa att att gga 3983 Thr Leu Mat Met Asp Asp Asp Ile Ser Lys His Leu Gln Ile Ile Gly agt gtt ctg gtt ccg gag gaa aga gaa gat aaa gaa aac atc atc aag 4031 Ser Val Leu Val Pro Glu Glu Arg Glu Asp Lys Glu Asn Ile Its Lys gca tgg ctc ttt tgc cat gag cag agg ata aga ttc att tat gga agg 4079 Ala Trp Leu Phe Cys His Glu Gln Arg Ile Arg Phe Ile Tyr Gly Arg gcc atg gag atg cca ttg gtt cta ccg tca gga ctc tgt gaa ctt tct 4127 Ala Met Glu Met Pro Leu Val Leu Pro Ser Gly Leu Cys Glu Leu Ser ett tct tca tgc agt att aca gat gas get tta get att tgc ctt ggt 4175 Leu Ser Ser Cys Ser Ile Thr Asp Glu Ala Leu Ala Ile Cys Leu Gly ggc etc act tca ctg aga act tta caa ttg aaa tat aat atg gca tta 4223 Gly Leu Thr Ser Leu Arg Thr Leu Gln Lau Lys Tyr Asn Met Ala Leu act aca ctt cca tca gaa aag gtg ttt gag cat ttg aca aag ctt gat 4271 Thr Thr Leu Pro 8er Glu Lys Val Phe Glu His Leu Thr Lys Leu Asp agg ttg gtt gta agt ggt tgt ttg tgt ctc aaa tca ctg ggg ggc tta 4319 Arg Leu Val val Ser Gly Cys Leu Cys Leu Lys Ser Leu Gly Gly Leu egt get get eca tct ett tcc tgt ttt aac tgt tgg gat tgt ect tct 4367 Arg Ala Ala Pro Ser Leu 9er Gars Phe Asn Cys Trp Asp Cys Pro Ser tta gag cta gca cgg gga gca gaa cta atg ccg ttg aac ctt get agc 4415 Leu Glu Leu Ala Arg Gly Als Glu Leu Met Pro Leu Asa Leu Ala Ser aat ctc agc atc ctt ggc tgc att ctt gca get gat tcg ttc att aat 4463 Asn Leu Ser Ile Leu Gly Cys Ile Leu Ala Ala Aap Ser Phe Ile Asn ggc ttg eca cat ctg aaa cat ctt tcc att gat gtc tgc aga tgc tcc 4511 Gly Leu Pro His Leu Lys His Leu Ser Ile Asp Val Cys Arg Cye Ser cca tcc tta tcg att ggc cac ctg acc tcc ctt gas tca tta tgt cta 4559 Pro Ser Leu Ser Ile Gly His Leu Thr Ser Leu Glu Ser Lau Cya Leu aat ggt ctc cct gat ctt tgc ttt gtt gaa ggc ttg tct tcc ctg cac 4607 Asn Gly Leu Pro Asp Leu Cys Phe Val Glu Gly Leu Ser Ser Leu His ctt aag cgc cta agt tta gta gat gtt gca aac ctc act gcc aag tgc 4655 Leu Lys Arg Leu Ser Leu Val Asp Val Ala Asn Leu Thr Ala Lys Cye atc tca ccg ttt cgt gtc cag gaa tcg ctc acg gtt agt agc tct gta 4703 Ile Ser Pro Phe Arg Val Gln Glu Ser Leu Thr Val Sar Ser Sar Val ttg ctc aac cac atg cta atg get gaa ggg ttt aca gcc cca cca aat 4751 Leu Leu Aan Hia Met Leu Met Ala Glu Gly Phe Thr Ala Pro Pro Asn ctt act ctt tta gat tgc aag gag ccg tca gtt tca ttt gaa gaa cct 4799 Leu Thr Leu Leu Asp Gyre Lys Glu Pro Ser Val Ser Phe Glu Glu Pro gca aat ctc tca tcc gtc aag cac ctg cac ttt tca tgt tgc gaa aca 4847 Ala Aen Leu Ser Ser Val LyB His Leu His Phe Ser Cys Cys Glu Thr 1205 lZlO 1215 gag tcc ctg cct aga aat cta aaa tct gtc tca agt ctg gag agt ctt 4895 Glu Ser Leu Pro Arg Asn Leu Lya Ser Val Ser Ser Leu Glu Ser Leu 12x0 12x5 1330 tct ata gaa cga tgc ccc aac ata gca tct tta cca gat ctg ccg tcc 4943 Ser Ile Glu Arg Cars Pro A~n Ile Als Ser Leu Pro Asp Leu Pro Ser tcc ctc cag cgc ata act ata ttg aat tgc ccc gtc ttg atg aag aat 4991 8er Leu aln Arg Its Thr Ile Leu Asn Cys Pro Val Leu bet Lys Asn tgc caw gaa cct gat gga gaa agc tgg cca wag att tcg cac gtt cgt 5039 Cys aln alu Pro Asp aly Gllu Ser Trp Pro Lys Ile Ser His Val Arg 1265 1x70 1275 1280 tgg wag agc ttt cca cca aaa tcg atc tgg ctt cct tagagttgcc 5085 Trp Lys 8er Phe Pro Pro Lys Ser Ile Trp Lsu Pro 1x85 1290 actttgaaat aaatgagaag gtacaggttc tactaattca ttttttccag cacaatttat 5145 gagtttctca atatttaaaa catttcatgt tctaaacagg caccttgacg tcacccctct 5205 tctcttgaag ctccagagtt caggctcaag tcagaagcca atccgtcgtt aatgctgtcg 5265 tccccgcggt tccctgtttt tgccgcttgt attgctccgc tacnnnngtt gctatatcat 53x5 tcattccttg gttgtgcaca attgccaata tgtatttctc tgacagaatg aagtgataac 5385 tgtggctagg gcttttgttt tcatgtgcac aattgctata tcattcatgt ggccaattgg 5445 attgattaca agtgtgcttg ctctattacc agttaaaagg tgattgcttg ttctttgtca 5505 ccaattggat tgattacaag tgtgcttgct ggtgtaagag atgaaaaggc cttgtatttt 5565 agtgcatcaa aactgaaggt ttttggtggt atgctccatt ctgttcaata tcctaaaccc 56x5 tacacacaaa atgtaaaacc ttacgcgctg gtttgcagca tctataacca aagcgtatca 5685 ttcattgatt gcacggcgct tctgcagcac ctatctgttg ntgctgctga tcaaccgctg 5745 tgatgttcaa caacttgtcc gggcgggcgt caaagctatt ctcactgagc aggctaccat 5805 aaacatcgac atccatctgn tggtgcaatg ctgattcaga agaagattgg aggagattaa 5865 atcacttctt aattcaaatt taacttaaaa tataasttsa atctctctaa tcccttcttg 5925 gattgnccta acccgacaac aaa 5948 <210> 2 <Z11> 1292 <Z12> PRT
<213> Zea sways <400> a Met Ala Asp Leu Ala Leu Ala Gly Leu Arg Trp Ala Ala Sar Pro Ile Val Asn Glu Leu Leu Thr Lys Ala Ser Ala Tyr Leu Ser Val Asp Mat ZO a5 30 Val Arg Glu Ile Gln Arg Leu Glu Ala Thr Val Leu Pro Gln Phe Glu Leu Val Ile Gln Ala Ala Gln Lys Ser Pro His Arg Gly Ile Leu Glu Ala Trp Leu Arg Arg Leu Lys Glu Ala Tyr Tyr Asp Ala Glu Asp Leu Leu Asp Glu His Glu Tyr Asn Val Lau Glu Gly Lys Ala Lys Ser Glu Lya Ser Leu Leu Leu Gly Glu His Gly Ser Ser Sar Thr Ala Thr Thr Val Met Lys Pro Phe His Ala Ala Met Ser Arg Ala Arg Aen Leu Leu 115 la0 1Z5 Pro Gln Asa Arg Arg Lau Ile Ser Lys Met Asn Glu Lau Lys Ala Ile Leu Thr Glu Ala Gln Gln Leu Arg Asp Leu Leu Gly Leu Pro His Gly Asn Thr Val Glu Trp Pro Ala Ala Ala Pro Thr Ser Val Pro Thr Thr Thr Ser Leu Pro Thr Ser Lys Val Phe Gly Arg Asp Arg Asp Arg Asp Arg Ile Val Asp Phe Leu Leu Gly Lys Thr Thr Thr Ala Glu Ala Ser Ser Ala Lys Tyr Ser Gly Lau Ala Ile Val Gly Leu Gly Gly Mat Gly Lys Ser Thr Leu Ala Gln Tyr Val Tyr Asn Asp Lys Arg Ile Glu Glu Cps Phe Asp Ile Arg Mat Trp Val Cys Ile Ser Arg Lys Leu Asp Val 245 a50 a55 His Arg His Thr Arg Glu I1~ Ile Glu Ser Ala Lys Lys Gly Glu Cys a6o ass a7o Pro Arg Val Asp Aen Leu Asp Thr Leu.Gln Gars Lys Leu Arg Asp Ile a75 a80 a85 Leu Gln Glu Ser Gla Lys Phe Lau Leu Val Lau Asp Asp Val Trp She a90 a95 300 Glu Lys Ser His Aen Glu Thr Glu Trp Glu Lau Pha Leu Ala Pro Leu 305 310 315 3a0 Val Ser Lys Gla Ser Gly Ser Lys Val Leu Val Thr Ser Arg Ser Lys 3a5 330 335 Thr Leu Pro Ala Ala Ile Cys Cys Glu Gln Glu His Val Ile His Leu Lys Aen Met Asp Asp Thr Glu Phe Leu Ala Leu Ph~ Lye His His Ala Phe Ser Gly Ala Glu Ile Lys Aep Gln Val Lau Arg Thr Lys Leu Glu Asp Thr Ala Val Glu Ila Ala Lys Arg Leu Gly Gln Cys Pro Leu Ala Ala Lys Val Leu Gly Ser Arg Lau ~s Arg Lys Lys Asp Ile Ala Glu Trp Lys Ala Ala Leu Lys Ile Gly Asp Leu Sar Asp Pro Phe Thr Ser 4a0 4a5 430 Leu Leu Trp Ser Tyr Glu Lys Leu Asp Pro Arg Leu Gla Arg Cys Phe Leu Tyr Cys Ser Lau Phe Pro Lys Gly His Arg Tyr Glu Ser Asa Glu Lau Val Hie Leu Trp Val Ala Glu Gly Phs Val Gly Sar Cys Asn Lou Ser Arg Arg Thr Leu Glu Glu Val Gly Met Asp Tyr Phe Asn Asp ltat Val 8er Val Ser Phe Phe Gla Leu Val Phe His Ile Tyr Cys Asp Sar Tyr Tyr Val Mat His Asp Ile Leu His Asp Phe Ala Glu Ser Leu Ser 515 520 5a5 Arg Glu Asp Cys Phe Arg Leu G1u Asp Asp Asn Val Thr Glu Ile Pro ors Thr Val Arg His Leu Ser Ile His Val Hie Ser Met Gln Lys His Lys Gln Ile Ile Cys Lys Leu His His Leu Arg Thr Ile Ile Cys Ile Asp Pro Leu Mat Asp Gly Pro Ser Asp Ile Phe Asp Gly Met Leu Arg Asn Gln Arg Lys Leu Arg Val Leu Ser Leu Ser Phe Tyr Asn Ser Lys Aen Leu Pro Glu Ser Ile Gly Glu Leu Lys His Leu Arg Tyr Leu Asn Lau Ile Arg Thr Leu Val Ser Glu Leu Pro Arg Ser Leu Cys Thr Leu Tyr His Leu Gln Leu Leu Trp Leu Asn His Mot Val Glu Asn Leu Pro Asp Lys Leu Gars Asn Lau Arg Lys Leu Arg His Leu Gly Ala Tyr Val Asn Asp Pha Ala Ila Glu Lys Pro Ile Cys Gln Ile Leu Asn Ile Gly Lys Leu Thr Sar Leu Gln His Ile Tyr Val Phe Ser Val Gln Lys Lys Gln Gly Tyr Glu Leu Arg Gln Leu Lys Asp Leu Asn Glu Leu Gly Gly 705 710 715 7a0 Ser Leu Lys Val Lys Asn Lsu Glu Asn Val Ile Gly Lys Asp Glu Ala Val Glu Ser Lys Leu Tyr Leu Lys Ser Arg Leu Lys Glu Leu Ala Leu Glu Trp Ser Ser Glu Asn Gly Met Asp Ala Met Asp Ile Leu Glu Gly la WO 99/45118 PCT/AU99l00130 Leu Arg Pro Pro Pro Gln Lau Ser Lys Lau Thr Ila Glu Gly Tyr Arg Ser Asp Thr Tyr Pro Gly Trp Lau Lau Glu Arg Ser Tyr Phe Glu Asn Leu Glu Ser Phe G1n Lau Ser Asn Cys Sar Leu Leu Glu Gly Lau Pro Pro Asp Thr Glu Leu Leu Arg Asn Cys Ser Arg Leu Arg Ila Asn phe Val Pro Asn Leu Lys Glu Leu Ser Asa Leu Pro Ala Gly Leu Thr Asp Leu Sar Ile Gly Trp Cys Pro Leu Leu Mat Phe Ile Thr Asn Aen Glu Lau Gly Gla His Asp Leu Arg Glu Asn Ila Ila Met Lys Ala Ala Asp Leu Ala Sar Lys Lau Ala Leu Met Trp Glu Val Asp Sar Gly Lye Glu Val Arg Arg Val Leu Phe Glu Asp Tyr Val Ser Leu Ile Arg Lau Met Thr Lau Mat Met Asp Asp Asp Ile Ser Lys His Lsu Gln Ila Ile Gly Ser Val Lau Val Pro Glu Glu Arg Glu Asp Lys Glu Asn Ile Ila Lys Ala Trp Leu Phe Cys His Glu Gln Arg Ile Arg Phe Ile Tyr Gly Arg Ala Met Glu Met 8ro Lsu Val Leu Pro Ser Gly Leu Cys Glu Lau Ser Leu Sar Ser qrs Sar Ile Thr Asp Glu Ala Leu Ala Ile Cys Leu Gly Gly Lau Thr Ser Leu Arg Thr Leu Gln Lau Lys Tyr Asn Met Ala Leu Thr Thr Leu Pro Sar Glu Lys Val Phe Glu His Lau Thr Lys Lau Asp Arg Lau Val Val Ser Gly Cys Leu Cys Leu Lya Ser Leu Gly Gly Leu Arg Ala Ala Pro Ser Leu Ser Gars Phe Asn Cys Trp Asp Cps Pro Ser Leu Glu Leu Ala Arg Gly Ala Glu Lau Met Pro Leu Asn Leu Ala Ser Asn Leu Ser Ile Leu Gly Cys Ile Leu Ala Ala Asp Ser Phe Ile Asn Gly Leu Pro His Leu Lye His Leu Ser Ile Asp Val Cys Arg Cys Ser Pro Ser Leu Ser Ile Gly His Leu Thr Ser Leu Glu Ser Leu Gars Leu Asn Gly Leu Pro Asp Leu Cys Phe Val Glu Gly Leu Ser Ser Leu His Leu Lys Arg Leu Ser Leu Val Asp Val Ala Asn Lau Thr Ala Lys Cys Ile Ser Pro Phe Arg Val Gla Glu 8er Leu Thr Val Sar Ser Ser Val Leu Leu Asn His Met Leu Met Ala Glu Gly Phe Thr Ala Pro Pro Asn Leu Thr Leu Leu Asp Cys Lys Glu Pro Ser Val Ser phe Glu Glu Pro Ala Asn Leu Ser Ser Val Lys His Leu His Phe Ser Cys Cys Glu Thr 1205 1x10 1x15 Glu Ser Leu Pro Arg Asn Leu Lys Ser Val Ser Ser Leu Glu Ser Leu m so laa5 la3o 8er Ile Glu Arg Cys Pro Asn Ile Ala Ser Leu Pro Asp Leu Pro Ser 1235 1x40 1x45 Ser Leu Gln Arg Ila Thr Ile Leu Asn Cys Pro Val Lau Met Lys Asn 1x50 1x55 1260 ors Gln Glu Pro Asp Gly Glu Ser Trp Pro Lys Ile Ser His Val Arg 1265 1x70 1x75 1x80 Trp Lys Ser Phe Pro Pro Lys Ser Ile Trp Leu Pro 1285 1x90 <a10> 3 <311> 4390 <212> DNA
<213> Zea ways <zao>
<aal> cDs <222> (262)..(4137) <400> 3 gcccgacgga gctgcttccc tccttgctgt gctgtgcttg ttccagattc tactccatac 60 gcccttgatt tattttactt cgtgtaagat ctccaatctc cacttttttc gcttgctgtg 120 ttagctacta tttccccttt gtgttgctgg atctggcgag asagaaaaat aattggtttg 180 tgcttcattt tttcattcag attcattaat tttatgtgtc tgcagagaaa aaagaaaaaa 240 aaaagcttct tactgaattt c atg gcc gac ttg gcg ctc gcc ggc tta agg 291 Net Ala Asp Leu Ala Lau Ala Gly Lsu Arg tgg gca gca tcg ccg att gtc aac gag ctt ctt act aaa get tca get 339 Trp Ala Ala Ser Pro Ile Val Asa Glu Leu Leu Thr Lys Ala 8er Ala 15 a0 25 tac ctc agt gtg gac atg gtg cgt gag atc caa cga cta gaa gcc act 387 Tyr Leu Ser Val Asp Het Val Arg Glu Ile Gln Arg Leu Glu Ala Thr gtc ctg cca cag ttc gag ctg gtg att caa gcg gcc cag aag agc ccc 435 Val Leu Pro Gln Phe Glu Leu Val Ile Gln Ala Ala Gln Lys Ser Pro cac agg ggc ata ctg gag gca tgg ctc cgg cgt ctc aaa gaa gcc tac 483 Hie Arg Gly Ile Leu Glu Ala Trp Lau Arg Arg Leu Lys Glu Ala Tyr tat gat gcc gag gac ttg ttg gsc gag cat gag tac aat gtc ctt gag 531 Tyr Asp Ala Glu Aap Leu Leu Asp Glu His Glu Tyr Aan Val Leu Glu ggc aag gcc aag agc gaa aaa agt ctc ctg ctg gga gag cat gga agc 579 Gly Lys Ala Lya Ser Glu Lys Ser Leu Leu Leu Gly Glu His Gly Ser tcc tcc act gca act act gtc atg aag cct ttt cat get get atg agc 6Z7 Ser Ser Thr Ala Thr Thr Val Met Lys Pro Phe Hia Ala Ala Met Ser agg gca cgg aac ttg ctc cct caa aac aga agg cta att agc aag atg 675 Arg Ala Arg Asn Leu Leu Pro Gln Asn Arg Arg Leu Ile Ser Lys Met 1~5 130 135 aac gag ctc aaa gca atc ctg aca gaa gcc caa caa ctt cga gat ctt 7Z3 Asn Glu Lau Lys Ala Ile Leu Thr Glu Ala Gln Gln Leu Arg Asp Leu ctt ggt ttg cca cat ggc aat acc gtc gag tgg cca get gca gca cct 771 Leu Gly Leu Pro His Gly Asa Thr Val Glu Trp Pro Ala Ala Ala 8ro 155 lsa lss 170 acc agt gtt ccc aca acc aca tca ctt ccc act tcc aag gtt ttt ggt 819 Thr Ser Val Pro Thr Thr Thr Ser Leu Pro Thr Ser Lys Val Phe Gly cgc gac agg gat cgt gat cgt ata gta gat ttt ctt ctc ggc aag aca 867 Arg Asp Arg Asp Arg Asp Arg Ile Val Asp Phe Leu Leu Gly Lye Thr aca act get gag gca agc tca get aag tac tcg ggt ttg gcc att gtt 915 Thr Thr Ala Glu Ala Ser Ser Ala Lys Tyr Ser Gly Leu Ala Ile Val Z05 210 al5 gga ttg gga gga atg ggg aag tcc acc tta gca cag tat gtc tat aat 9fi3 Gly Lau Gly Gly Met Gly Lys Ser Thr Leu Ala Gla Tyr Val Tyr Asn Z20 225 a30 gac aaa agg ata gaa gaa tgc ttt gat atc agg atg tgg gtg tgc atc 1011 Aep Lys Arg Ile Glu Glu Cys Phe Asp Ile Arg Met Trp Val Cars Ile 235 a40 a45 X50 tca cgc aaa ctt gat gtg cat cgt cac aca agg gag att ata gag tct 1059 Ser Arg Lye Leu Asp Val His Arg His Thr Arg Glu Ile Ile Glu Ser gca aaa aag gga gag tgc cca cgt gtt gat aat ctc gat act ctc cag 1107 Ala Lys Lys Gly Glu Cys Pro Arg Val Asp Asn Leu Asp Thr Leu Gln z7o a75 aeo tgc aaa tta cgt gat ata cta caa gag tca cag aaa ttc ctg ctt gtc 1155 Cys Lys Leu Arg Asp Ile Leu Gln Glu Ser Gln Lys Phe Leu Leu Val 285 a90 Z95 ttg gat gat gtt tgg ttt gaa aaa tct cat aat gag aca gag tgg gag 1203 Leu Asp Asp Val Trp Phe Glu Lys Ser His Asn Glu Thr Glu Trp Glu tta ttc ctt get cca tta gtc tat aaa cag tca ggg agc aaa gtt ttg 1x51 Leu Phe Leu Ala Pro Leu Val Ser Lys Gln Ser Gly Ser Lys Val Leu gtg act tct cga agt aaa aca ctt cct gcc get att tgt tgt gaa caa 1299 Val Thr Ser Arg Ser Lys Thr Leu Pro Ala Ala Ile Gars Cys Glu Gln gaa cat gtc att cat ttg aaa sac atg gat gat act gag ttt ttg get 1347 Glu His Val Ile His Leu Lys Asn Met Asp Asp Thr Glu Phe Leu Ala ctt ttt aaa cac cat get ttc tct gga gca gaa atc aaa gac caa gta 1395 Leu Phe Lys His His Ala Phe Ser Gly Ala Glu Ile Lys Asp Gln Val tta cgc acg aag ctg gaa gac act gca gtg gag att get aaa agg ctt 1443 Leu Arg Thr Lys Leu Glu Asp Thr Ala Val Glu Ile Ala Lys Arg Leu gga caa tgt cct ttg gca gca aaa gtt ctg ggt tct cga ttg tgc agg 1491 Gly Gln Cys Pro Leu Ala Ala Lys Val Leu Gly Ser Arg Leu Cys Arg aaa aag gat att get gaa tgg aaa get get cta aag att gga gat tta 1539 Lys Lys Asp Ile Ala Glu Trp Lys Ala Ala Leu Lys Ile Gly Asp Leu 415 4a0 4a5 agt gat ccc ttc aca tct ctg ttg tgg agt tac gag aag tta gat cca 1587 Ser Asp Pro She Thr Ser Leu Leu Trp Ser Tyr Glu Lye Leu Asp Pro cgt ctg cag agg tgc ttc ttg tat tgc agc ttg ttt cca aaa ggt cat 1635 Arg Leu Gln Arg Cys Phe Lsu Tyr Cys gar Lau Phe Pro Lys Gly His aga tat gaa tct aat gag ttg gtt cac ctt tgg gtg gca gaa gga ttt 1683 Arg Tyr Glu Ser Asn Glu Leu Val His Lau Trp Val Ala Glu Gly Phe gtt ggt tca tgc aat ttg agt agg aga acg tta gaa gag gtt ggg atg 1731 Val Gly Ser Cyrs Aan Lau Ser Arg Arg Thr Leu Glu Glu Val Gly Met gat tac ttc aat gat atg gtc tct gta tct ttc ttc caa ttg gtt ttt 1779 Asp Tyr Phe Asn Aap Met Vsl Ser Val 8er Phe Phe Gln Leu Val Phe cat atc tat tgt gat tcg tac tat gtc atg cat gat atc ctt cat gat 1827 Hia Ile Tyr Cys Asp Sex Tyr Tyr Val Met His Aap Ile Leu Hia Aap ttt gca gag tca ctc tat aga gaa gac tgc ttt aga tta gaa gat gat 1875 Phe Ala Glu Ser Leu 8er Arg Glu Asp Cys Phe Arg Leu Glu Asp Asp aat gtg aca gaa ata cca tgc act gtt cga cat cta tct att cat gtt 1923 Asn Val Thr Glu Ile Pro Cys Thr Val Arg His Leu Ser Ile His Val cat agt atg caa aag cat aag caa att atc tgc aag cta cat cat tta 1971 His Ser Met Gln Lye His Lya Gln Ile Ile Cys Lya Leu His Hie Leu cgc act att atc tgc atc gat ccg cta atg gat ggc cca agt gat att 2019 Arg Thr Ile Ile Cys Ile Asp Pro Leu Met Asp Gly Pro Ser Aap Ile ttt gat ggc atg cta cgg aac caa aga aaa ctg cgt gta ttg tct ctg 2067 Phe Asp Gly Mat Leu Arg Asn Gln Arg Lys Leu Arg Val Leu Ser Leu tca ttt tac aac agc aaa aat ttg cca gaa tct att ggt gag ctg aag 2115 Ser Phe Tyr Aan Ser Lys Aan Leu Pro Glu Sar Ile Gly Glu Leu Lys cac ctc cgg tat ttg aac ctc atc agg acg tta gtt tct gaa ttg cct 2163 His Leu Arg Tyr Leu Aan Leu Ile Arg Thr Leu Val 8er Glu Leu pro aga tca tta tgt act ctc tac cac tta caa tta ctt tgg tta aac cac 2211 Arg Ser Leu Cya Thr Leu Tyr His Leu Gln Leu Leu Trp Leu Asn His atg gtg gag aat ttg cct gac aaa cta tgc aat tta aga aag cta cga 2259 Met Val Glu Asn Leu Pra Asp Lys Lau Cys Asn Leu Arg Lye Leu Arg cat cta gga gcg tac gtg aat gat ttc gcg att gaa aag cct stt tgc 2307 His Leu Gly Ala Tyr Val Asn Asp Phe Ala Ile Glu Lys Pro Ile Cys caa att ctg aat ata ggt aag tta acg tcg cta caa cac att tat gtc x355 Gln Ile Leu Asn Ile Gly Lys Leu Thr Ser Leu Gln His Ile Tyr Val ttt tct gta caa aag aag caa ggc tat gag ttg cga cag ttg aag gac 2403 Phe Sex Val Gln Lys Lys Gln Gly Tyr Glu Leu Arg Gln Leu Lys Asp ttg aat gag ctt ggt ggc agt tta aaa gtg aaa aat ctt gag aat gtc 2451 Leu Asn Glu Leu Gly Gly Ser Leu Lys Val Lys Asn Leu Glu Asn Val ?15 720 725 730 att gga aag gat gaa gcc gta gag tcg aag cta tat ctg aaa agt cgc 2499 Ile Gly Lys Asp Glu Ala Val Glu $er Lys Lau Tyr Leu Lys Ser Arg ctt aaa gag ttg gca ctt gag tgg agt tcc gag aat gga atg gat gca 2547 Leu Lya Glu Leu Ala Leu Glu Trp Ser $er Glu Asa Gly Met Asp Ala atg gat att cta gaa ggt ctg aga cca cca ccc cas ctg agt aag ctc 2595 Diet Asp Ile Leu Glu Gly Leu Arg Pro Pro Pro Gln Leu $er Lys Leu aca atc gaa ggt tac aga tct gat aca tat cct ggg tgg tta cta gag 2643 Thr Ile Glu Gly Tyr Arg $er Aap Thr Tyr Pro Gly Trp Leu Leu Glu cga tcc tat ttt gag aat ttg gaa agt ttt cag ctt agt aat tgc agt x691 Arg Ser Tyr She Glu Asn Leu Glu Ser Phe Gln Lau $er Asn Cys Ser ttg cta gaa ggc cta cca cca gat aca gag ctc ctt cgg ant tgc tct 2739 Leu Leu Glu Gly Leu Bro Pro Asp Thr Glu Leu Leu Arg Asn Cars Ser 815 8a0 825 agg ttg cgt ata aac ttt gtt cca aat ttg aag gaa cta tct sat ctt x787 Arg Leu Arg Ile Asn Phe val pro Asn Leu Lys Glu Leu Ssr Asn Leu cca gca ggc ctt aca gat tta tca att ggt tgg tgc cca ctg ctt atg 2835 Pro Ala Gly Leu Thr Asp Leu Ser Ile Gly Trp ~s Pro Leu Leu Met ttt atc acc aac aat gag cta gga cag cat gac ttg agg gaa aat ata 2883 Phe Ile Thr Asn Asn Glu Leu Gly Gln His Asp Leu Arg Glu Asn Ile ata atg aag gca gcc gac ctg gca tct aaa ctt gca ttg atg tgg gag 2931 Ile Met Lya Ala Ala Asp Leu Ala Ser Lys Leu Ala Leu Met Trp Glu gtg gat tca gga aaa gaa gtt agg aga gta ctg ttt gaa gac tat gta 2979 Val Asp Ser Gly Lya Glu Val Arg Arg Val Leu Phe Glu Asp Tyr Val tct ctg att cgg ttg atg aca ttg atg atg gat gat gat ata tca aag 3027 Ser Leu Ile Arg Leu Met Thr Leu Met Met Asp Aap Asp Ile Ser Lye cat ctt caa att att gga agt gtt ctg gtt ccg gag gaa aga gaa gst 3075 His Leu Gln Ile Zle Gly Ser Val Leu Val Pro Glu Glu Arg Glu Asp asa gas aac atc atc aag gca tgg ctc ttt tgc cat gag cag agg ata 3123 Lys Glu Asn Ile Ile Lys Ala Trp Leu Phe Cys His Glu Gln Arg Ile aga ttc att tat gga agg gcc atg gag atg cca ttg gtt cta ccg tca 3171 Arg Phe Ile Tyr Gly Arg Ala Met Glu Mat Pro Leu Val Leu Pro Ser gga ctc tgt gaa ctt tct ctt tct tca tgc agt att aca gat gaa get 3219 Gly Leu Cya Glu Leu Ser Leu Ser Ser Cys Ser Ile Thr Aap Glu Ala tta get att tgc ctt ggt ggc ctc act tca ctg aga act tta caa ttg 3267 Leu Ala Ile bra Leu Gly Gly Leu Thr Ser Leu Arg Thr Leu Gln Leu aaa tat aat atg gca tts act sca ctt cca tca gaa aag gtg ttt gag 3315 Lya Tyr Asn Met Ala Leu Thr Thr Leu Pro Ser Glu Lya Val Phe Glu cat ttg aca aag ctt gat agg ttg gtt gta agt ggt tgt ttg tgt ctc 3363 His Leu Thr Lys Leu Asp Arg Leu Val Val Ser Gly Cys Leu Cys Leu loco loa5 1030 aaa tca ctg ggg ggc tta cgt get get cca tct ctt tcc tgt ttt aac 3411 Lys Ser Leu Gly Gly Lau Arg Ala Ala Pro Sar Leu Ser Cys Phe Aan tgt tgg gat tgt cct tct tta gag cta gca cgg gga gca gaa cta atg 3459 Cys Trp Asp Cys Pro Ser Leu Glu Leu Ala Arg Gly Ala Glu Leu Mat ao ccg ttg aac ctt get agc aat ctc agc atc ctt ggc tgc att ctt gca 3507 Pro Leu Asa Leu Ala Ser Asa Leu Ser Ile Leu Gly Cys Ile Leu Ala get gat tcg ttc att aat ggc ttg cca cat ctg aaa cat ctt tcc att 3555 Ala Aep Ser Phe Ile Asn Gly Leu Pro His Leu Lys His Leu Ser Ile gat gtc tgc aga tgc tcc cca tcc tta tcg att ggc cac ctg acc tcc 3603 Asp Yal Cys Arg Cys Ser Pro Ser Leu 8er Ile Qly His Leu Thr Ser ctt gaa tca tta tgt cta aat ggt ctc cct gat ctt tgc ttt gtt gaa 3651 Leu Glu Ser Leu Cys Leu Asn Gly Leu Pro Asp Leu Cys Phe Val Glu ggc ttg tct tcc ctg cac ctt aag cgc cta agt tta gta gat gtt gca 3699 Gly Leu Ser Ser Leu His Leu Lys Arg Leu Ser Leu Val Asp Val Ala aac ctc act gcc aag tgc atc tca ccg ttt cgt gtc cag gaa tcg ctc 374'1 Asn Leu Thr Ala Lys Cys Ile Ser Pro Phe Arg Val ala alu Ser Leu acg gtt agt agc tct gta ttg ctc aac cac atg cta atg get gaa ggg 3795 Thr Val Ser Ser Ser Val Leu Leu Asn His Met Leu Met Ala Glu <'ily ttt aca gcc cca cca aat ctt act ctt tta gat tgc aag gag ccg tca 3843 Phe Thr Ala Pro Pro Asn Leu Thr Leu Leu Asp Cys Lys Cilu Pro 8er gtt tca ttt gaa gaa cct gca aat ctc tca tcc gtc sag cac ctg cac 3891 Val Ser Phe Glu Glu Pro Ala Asn Lsu Ser Ser Val Lys His Leu His ttt tca tgt tgc gaa aca gag tcc ctg cct aga aat cta aaa tct gtc 3939 Phe Ser Cars Cys dlu Thr Glu Ser Leu Pro Arg Asa Leu Lys Ser Val tca agt ctg gag agt ctt tct ata gaa cga tgc ccc aac ata gca tct 3987 Ser Ser Leu alu Ser Leu Ser Ile Glu Arg Cys Pro Asa Ile Ala 8er tta cca gat ctg ccg tcc tcc ctc cag cgc ata act ata ttg aat tgc 4035 Leu Pro Asp Leu Pro 8er Ser Leu Gln Arg Ile Thr Ile Leu Asa Cps al 1245 1x50 1x55 ccc gtc ttg atg aag aat tgc cas gaa cct gat gga gaa agc tgg cca 4083 Pro Val Leu Met Lys Asn Cys Gla Glu Pro Asp Gly Glu Ser Trp Pro aag att tcg cac gtt cgt tgg aag agc ttt cca cca aaa tcg ate tgg 4131 Lye Ile Ser His Val Arg Trp Lya 8er Phe Pro Pro Lys Ser Ile Trp 1275 1280 1285 1x90 ctt cct tagagttgcc actttgaaat aaatgagaag gcaccttgac gtcacccctc 4187 Leu Pro ttctcttgaa gctccagagt tcsggctcaa gtcagaagcc aatccgtcgt taatgctgtc 4x47 gtccccgcgg ttccctgttt ttgccgcttg tattgctccg ctacnnrusgt tgctatatca 4307 ttcattcctt ggttgtgcac aattgccaat atgtatttct ctgacagaat gaagtgataa 4367 ctgtggctag ggcttttgtt ttc 4390 <210> 4 <211> 1292 <ala> pRT
<213> Zea ways <400> 4 Met Ala Asp Leu Ala Leu Ala Gly Leu Arg Trp Ala Ala Ser Pro Ile Val Asn Glu Leu Leu Thr Lys Ala Ser Ala Tyr Leu Ser Val Asp Met Val Arg Glu =le Gln Arg Leu Glu Ala Thr Val Leu Pro Gln Phe Glu Lau Val Ile Gln Ala Ala Gln Lys Ser Pro His Arg Gly Ile Leu alu Ala Tip Leu Arg Arg Leu Lys Glu Ala Tyr Tyr Asp Ala Glu Asp Leu Leu Asp Glu His Glu Tyr Asn Val Leu Glu Gly Lys Ala Lys Ser alu Lys Ser Leu Leu Leu Gly Glu His Gly Ser Ser Ser Thr Ala Thr Thr Val Met Lye Pro Phe His Ala Ala Mot Ser Arg Ala Arg Asa Leu Leu Bro Gla Asn Arg Arg Leu Ile Ser Lys Met Asa Glu Leu Lys Ala Ile Leu Thr Glu Ala Gln Gln Leu Arg Asp Leu Lsu Gly Leu Pro His Gly Asn Thr Val Glu Trp Pro Ala Ala Ala Pro Thr Ser Val Pro Thr Thr Thr Ser Leu Pro Thr Ser Lys Val Bhe Gly Arg Asp Arg Asp Arg Asg Arg Ile Val Asp Phe Lau Leu Gly Lys Thr Thr Thr Ala Glu Ala Ser Ser Ala Lys Tyr Ser Gly Leu Ala Ile Val Gly Leu Gly Gly Met Gly Lys Ser Thr Leu Ala Gln Tyr Val Tyr Aen Asp Lye Arg Ile Glu (31u Cys Phe Asp Ile Arg Met Trp Val Cys Ile Ser Arg Lys Leu Asp VaI

His Arg His Thr Arg Glu Ile Ile Glu Ser Ala Lys Lys Gly Glu Cys Pro Arg Val Asp Asn Leu Asp Thr Leu Gln Cys Lys Leu Arg Asp Ile Leu Gln Glu Ser Gln Lys Phe Leu Lau Val Leu Asp Asp Val Trp Hhe Glu Lys Ser His Aen Glu Thr Glu Trp Glu Leu Phe Leu Ala Pro Lau Val Ser Lys Gln Ser Gly Ser Lys Val Leu Val Thr Ser Arg Ser Lys Thr Leu Pro Ala Ala Ile Cys ors Glu Gla Glu His Val Ile His Leu Lys Asn Met Asp Asp Thr Glu Bhe Leu Ala Leu Phe Lys His His Ala Phe Ser Gly Ala Glu Ile Lys Asp Gln Val Leu Arg Thr Lys Leu Glu Asp Thr Ala Val Glu Its Ala Lys Arg Leu Gly Gln Cys Pro Lau Ala Ala Lys Val Leu Gly Ser Arg Lsu Cys Arg Lys Lye Aap Ile Ala Glu Trp Lys Ala Ala Leu Lys Ile Gly Asp Lsu Ser Asp Pro Phe Thr Ser Leu Leu Trp Ser Tyr Glu Lys Leu Asp Pro Arg Leu Gla Arg Cys Phe Leu Tyr Cys Ser Leu Phe Pro Lys Gly His Arg Tyr Glu Ser Asn Glu Leu Val His L~u Trp Val Ala Glu Gly Phe Val Gly Ser Cys Asn Lau Ser Arg Arg Thr Leu Glu Glu Val Gly Met Asp Tyr Phe Asn Asp Met Val Ser Val Ser Phe Phe Gln Leu Val Phe His Ile Tyr Cys Asp Ser Tyr Tyr Va1 Met His Asp Ile Lsu His Asp Phe Ala Glu Ser Leu Ser Arg Glu Asp Cys Phe Arg Leu Glu Asp Asp Asn Vsl Thr Glu Ile Pro Cys Thr Val Arg His Leu Ser Ile His Val His eer Met Gln Lys His Lye Gln Ile Ile Cys Lys Leu His His Leu Arg Thr Ila Ile Cys Ile Asp Pro Leu Met Asp Gly Pro Sar Aap Ile Phe Asp Gly Met Leu Arg Asn Gln Arg Lys Leu Arg Val Leu Ser Leu Sar Phe Tyr Asn Ser Lys Asn Leu Pro Glu Ser Ile Gly Glu Leu Lys His Leu Arg Tyr Leu Asa a4 Leu Ile Arg Thr Leu Val Ser Glu Leu Pro Arg Ser Leu Cya Thr Lau 6a5 630 635 ~ 640 Tyr His Leu Gln Leu Leu Trp Leu Aan Hia Mat Val Glu Aen Leu Pro Aap Lye Leu Cys Aan Leu Arg Lya Leu Arg His Leu Gly Ala Tyr Val Aan Asp Phe Ala Ile Glu Lye Pro Ile Cys Gln Ile Leu Aan Ile Gly Lys Leu Thr Ser Leu Gln His Ile Tyr Val Phe Ser Val Gln Lya Lys Gln Gly Tyr Glu Leu Arg Gln Leu Lys Asp Leu Asn Glu Leu Gly Gly Ser Leu Lya Val Lys Asn Leu Glu Aan Val Ile Gly Lya Asp Glu Ala Val Glu Ser Lye Leu Tyr Leu Lye Ser Arg Leu Lya Glu Lau Ala Leu Glu Trp Ser Ser Glu Asn Gly Met Asp Ala Met Asp Ile Leu Glu Gly 755 ?60 765 Leu Arg Pro Pro Pro Gln Leu Ser Lya Leu Thr Ile Glu Gly Tyr Arg Ser Aap Thr Tyr Pro Gly Trp Leu Leu Glu Arg Ser Tyr Phe Glu Asn Leu Glu Ser Phe Gln Leu Ser Aan Cya Ser Leu Leu Glu Gly Leu Pro Pro Asp Thr Glu Leu Leu Arg Asn bra Ser Arg Leu Arg Ile Asn Phe Sao 8z5 830 Val Pro Asn Leu Lya Glu Leu Ser Asa Leu Pro Ala Gly Leu Thr Asp Leu Ser Ile Gly Trp Cys Pro Leu Leu Met Phe Ile Thr Asn Aan Glu Leu Gly Gln His Aap Leu Arg Glu Asn Ile Ile Mat Lys Ala Ala Aap Leu Ala Ser Lye Leu Ala Leu Met Trp Glu Vsl Asp Ser Gly Lys Glu 885 890 ~ 895 Val Arg Arg Val Leu Phe Glu Asp Tyr Val Ser Leu Ile Arg Leu Met Thr Leu Met Met Asp Asp Asp Ila Ser Lys His Leu Gln Ile Ile Gly Ser Val Leu Val Pro Glu Glu Arg Glu Asp Lys Glu Asn Ile Ile Lys Ala Trp Leu Phe Cys His Glu Gln Arg Ile Arg Phe Ile Tyr Gly Arg Ala Met Glu Met Pro Leu Val Leu Pro Ser Gly Leu Cys Glu Leu Ser Leu Sar 8er Gys Ser Ile Thr Asp Glu Ala Leu Ala Ile Gys Leu Gly Gly Lsu Thr Ser Lau Arg Thr Leu Gln Leu Lys Tyr Asa Met Ala Leu Thr Thr Leu Pro Ser Glu Lys Val Phe Glu His Leu Thr Lys Leu Asp Arg Leu Val Val Ser Gly Cys Leu Cys Leu Lys Ser Leu Gly Gly Leu Arg Ala Ala Pro Ser Leu Ser Cys Phe Asn Cys Trp Asp Cys Pro Ssr Leu Glu Leu Ala Arg Gly Ala Glu Leu Met Pro Leu Asn Leu Ala Ser Asn Leu Ser Ile Leu Gly Cys Ile Leu Ala Ala Asp Ser Phe Ile Aen Gly Leu Pro His Leu Lys His Leu Ser Ile Asp Val Cars Arg ors Ser Pro Ser Leu Ser Ile Gly His Leu Thr Ser Leu Glu Ssr Leu Cys Leu Asn Gly Leu Pro Asp Leu Gars Phe Val Glu Oly Leu Sar Sar Leu His Leu Lys Arg Leu Ser Leu Val Asp Val Ala Asa Lsu Thr Ala Lys Gars Ile Ser Pro Phe Arg Val Gla Glu Sar Leu Thr Val Bar Sar Ser Val Lau Leu Asn His Met Leu Met Ala Glu Gly Phe Thr Ala Pro Pro Asn Leu Thr Leu Leu Asp Cys Lys Glu Pro Ser Val Ser Phe Glu Glu Pro Ala Asn Leu Ser S~r Val Lys His Leu His Phe Ser Cys Cys Glu Thr 1205 1x10 1x15 Glu Ser Leu Pro Arg Asn Lau Lys Ser Val Ser Ser Leu Glu S~r Leu 1220 1225 1x30 Ser Ile Glu Arg Cys Pro Asn Ile Ala Ser Leu Pro Asp Leu Pro Ser Ser Leu Gln Arg Ile Thr Ila Leu Asa Cys Pro Val Leu Met Lys Asn 1x50 ~ 1255 1260 Cys Oln Glu Pro Asp Gly Glu Sar Trp Pro Lys Ila Ser His Val Arg Z65 1x70 1275 1280 Trp Lys Ser Phe Pro Pro Lys Sar Ile Trp Leu Pro <Z10> 5 <all> 8205 <212> DNA
<a13> Hordeum sp.
<ZZO>
<aal> cns <Z22> (3767)..(7648) <400> 5 gaattcatac atgtaggtgg agatatatta aaagctatgt gtagtgtgcg ccaaagaagt 60 tttgcaagcg ggcattctaa gaaaagatgt ttaatcgttt cattttgctc acaaaaacaa 1a0 caattcaccc tgcctctcca tcttcgtttt ttcsaattat ctttggttaa taccacaccc 180 a7 ttgtgtatga accacatgaa aactttaatc ctcaggggta ccttgatttt ccatatatgt Z40 agcgatctag atattagcgc attatcaatg agatccatgt acatagattt aactaagaat 300 actccgttgg aagccaaagt ccaacgaata gtatgcgtct ggttagatag ttgaatctgc 360 attaaccttc tgaccagatg tagccatgtg ttccaccgtg gtccaactaa agctctccta 420 aattgttgtt aagaggtatg gattgaagta ctgtgtcaac ataatcctct tttcgttggg 480 caatgttata tagagttgga tattgaagag ctaggggagt gtctcccaac caagtatcct 540 cccaaaacct tgttgatgca ecatccccaa ctacaaatct tacccgctgg aaaaaggtca 600 cctttacctt cattagacct ttccagaaag gagaatcctg tggtctaact ttaacctgtg 660 ctaaagtttt tgaatgtaag tatttatttt taagtatttg aaatcacatt ccttcagttt 720 tcatagatag cctaaaaagc catttgctga gaaggcatct gttcttgact tgtaagtttt 780 caatacccat acccccctgt tctttaggtc tacatataat atcccaacgt gtcagtctgt 840 atttcttctt gacctcgtca cttttccaga agaaccgaga cctataaaaa tctagtcttt 900 ttcgaacccc tactggtatt tcaaaaaaag acaagagaaa catgggcsgg ctagttaaca 960 ccgaatttat taatactggc cttcctccat atgacataag cttgcccttc caacaactta 1020 gtttttttta aaatctatct tcaatacatt tccactcctt attcgtcagt cgcctatgat 1080 gaatagggat cccaagataa ctaaatggta gtgatcccat ttcacatcca aaaagctgtc 1140 tataagcgtc ctgttcctct tttgcctttc caaagcaaaa tagttcactt ttgttgaaat 1200 ttatttttaa tcctgataat tgttcaaaaa gacacaataa tagtttcatg ttccttgctt 1260 tggccatatc gtgctccatg sasatsattg tatcatccgc gtattgtaaa stagacaccc 13x0 ctccttctac aagattcggt atgaggcctc caacttggcc tttttctttg gctctttgta 1380 ttaatattgt taacatgtct gcaactatgt taaacaacac tggggataat gaatcacctt 1440 gcctgagtcc cttatgtgtt tggaaaaaat gacctatgtc atcattsact ttaacaccta 1500 cactcccttt ttggstgaaa ttatcaacct gatttctcca tgcatcattg aatcctttca 1560 tacgtaaggc ctgttggaga aaagaccatt ttactttatc atatgctttc tcgaaatcca 1620 as ctttaaatat aaccccatct agtttctttg tatggatttc atgcaatgtc tcatggaggs 1680 ccacaacacc ctccaagata tgtctcccag gcatgaacgc agtctgcgtc ggttggacta 1740 cagaattagc aatctttgtt aatctgtttg taccaacttt tgtaaagatc ttgaaactca 1800 cgttgsgcag acatatcggt ctaaactgtt caattcgaat agcttccacc ttttttggca 1860 acagtgtaat tgttccaaag ttgagatgaa ataattgcaa gtgaccatta aatsgatcat 19x0 gaaacatatg catcaagtca ctcttaataa tgtaccaaca cttcttataa aactcagcag 1980 ggaacccatc tggtcccggt gctttattta atttcatttg tgtgattgag tcatatactt 2040 ctttctctgt aaatggagct gataacacct cattctcatc tgcatttagt tgaggtatat 2100 catgagtgat gctttcatcc aaagagacca ttgtctcttc tggtttagca aataattttt x160 tgtaaaattc agagatatat aattttaagt tctcgtgtcc tataattgtt ccttcatctt 2220 gctcaagctg agtaattttt ttcttcctgt gtttgccatt tgcaatcatg tggaagaatt 2280 gtgtgttatc atccccatga accacctttg ccaccttggc tccagttgcc catttcatct x340 cctcttctct taggagtttt tgcaacctcc tctcagcatc satcttttta ttcctgtccg 2400 tcaaattgag ttgattagac tccgctttaa tgtctaattc attaatcaga tgacagagta 2460 gttctttttc aattctatat ttaccactct gattttttgc ccatcctctt aaaaactgtc 2520 gtaaatttct aattttgttt tgccatstat cgatatttgt tatcccaccc gagtcccttg 2580 cccattcttc ggctatgaga tccatgaaac cttctctttc aaaccaactt aattcaaaag x640 aaaagatatt tctatttccc acatgatttg cctctccgga gtccagtagt agaggcgtgt 2700 gatcagatat agctctttgc aaggcctgta ctgtaaccag tgggtatttc tgttcccact 2760 caacgctagt cagtacccgg tccaatttct catatgttgg taccggtaaa gaatttgccc 2820 aagtaaattg tctaccagtg agatcaatct ccctcaaatt aagactctca atgaccatgt 2880 taaacatagt ggaccagcgt gtatcaaaat tatcattgtt tttttcattt tgtcttctaa 2940 tgatgttaaa gtctcccccc ccataagtgg taatctttcg tctccacaaa tccgtaccaa 3000 atctgccaga aaatgaggtt tgagttctgg ttgcgctgcc ccatatacag ccaccagagc 3060 ccacctgaac ccatcggtct tcgacctcag tcgaaattta actgtgaact cgccatatat 3120 cacattcatc acctctagtg tttcacattt taccccaagt aagatcccac csgatcttcc 3180 ccttggcggt aaacaatgcc agtcaaaatc aacccctccc gagaggatat tsagaaattg 340 tgatgtgaaa ttatccctac ccgtttcttg taaagctata aagtcaagtt tatgttcgag 3300 agatgcttct ctaaggaacc ttcttttagc caagtccgct agacctctgc tattccaaaa 3360 aatccctttc atatctcatc atgaaatttt ttcttttgtt ttactctggc actcctcccg 34x0 accgctgatt gaggaaatac ttttctattc caaggtctct tgggtttgtc tttaatacca 3480 tctaagttat tttgtactac tctgacattt ggaagacaac cgtcaatcga aggagccaca 3540 tacccctccg atcccatgtc atccagatct acctcatctt cctctagatg tagtaaattg 3600 tcacaaagag tctgtagttg tgcacctcct aagtcattaa tatctgastc ttccattggt 3660 tttaatgctg ctaaatgctt tgtctttctg cacagaagat atgtgcatcc acasagcaaa 37x0 cacaactccc ctccctagga atatacacca aaagaactgc aatacg atg gca gaa 3775 Met Ala Glu gtg gcg tta get gga gca gcc tta agt gtt agc tta aat ttg gcc gca 3823 Val Ala Leu Ala Gly Ala Ala Lou Ser Val Bar Lau Asn Leu Ala Ala tcg cca gtc ttg aag aag ctc ctt get aat get tca ata tac ctt gga 3871 Ser Pro Val Leu Lys Lys Lsu Leu Ala Aan Ala Bar Ile Tyr Leu Gly a0 a5 30 35 gtg gac atg gca ctt gag ctc ctt gaa ctg gag aca act atc ttg cca 3919 Val Asp Mat Ala Leu Glu Leu Lou Glu Leu Glu Thr Thr Its Leu Bro cag ttc gag ctg gtg atc gaa gca gca aac aag gga aac cac agg ccc 3967 Gln Phe Glu Lau Val Ile Glu Ala Ala Asn Lys Gly Aen His Arg Pro aag ttg gac aaa tgg ctt cag gaa ctc aaa gaa ggc ttc tac ctg get 4015 Lye Leu Asp Lys Trp Leu Gln Glu Leu Lys Glu Gly Pha Tyr Leu Ala gaa gac ttg ttg gac gac cat gag tac aac ctc ctc aag cgc caa gca 4063 Glu Asp Leu Leu Asp Asp His Glu Tyr Asn Leu Leu Lys Arg Gln Ala aag gga aag gat cec ttg acg gcc aat ggc tcc tcc atc agc aac act 4111 Lye Gly Lye Asp Pro Leu Pro Ala Asn Gly Ser Ser Ile Ser Asn Thr ttt atg aag cct ctg cgt get gca tcg agc agg ctg tcg aat ttt agt 4159 Phe Met Lys Pro Leu Arg Ala Ala Ser Ser Arg Leu Ser Asn Phe Ser tca gag aac aga aag cta att caa cag tta aat ggg cta aag get acc 4207 Ser Glu Asa Arg Lys Lau Ile Gln Gln Leu Asn Gly Leu Lys Ala Thr ctg gcg aaa gcc aag gac ttc cgt gaa ctg ctc ttc tta cca tct ggt 4255 Leu Ala Lys Ala Lye Asp Phe Arg Glu Leu Leu Phe Leu Pro Ser Gly tat agt gca gag ggc tcc acc ata cca tca get gtt gtt cct gag act 4303 Tyr Ser Ala Glu Gly Ser Thr Ila Pro Ser Ala Val Val Pro Glu Thr agt tca ata gca cct ctg aaa gtg ata ggt cgt aga aag gat cgc aac 4351 Ser Ser Ile Ala Pro Leu Lys Val Ile Gly Arg Arg Lys Asp Arg Asn cat ata ata aat tgt ctt acc aag aca aca gtg act acc aag tct agt 4399 His Ile Ile Asn Cya Leu Thr Lys Thr Thr Val Thr Thr Lys Ser 8er aaa acc atg tac tcg ggt ttg get att gtt gga get gga ggc atc gga 4447 Lys Thr Met Tyr Ser Gly Leu Ala Ile Val Gly Ala Gly Gly Ile Gly a15 220 2a5 aag tec agc ttg gca cag ctt gtt tac aat agc aag agg gta aaa aaa 4495 Lya Ser Ser Leu Ala Gln Leu Val Tyr Asn Ser Lys Arg Val Lys Lys cat ttt aat gtg aga atg tgg sta agc ata tca cgc aaa ctt gat gtt 4543 His Phe Asn Val Arg Met Trp Ile 8er Ile Ser Arg Lys Leu Asp Val cga cgt cat aca cgg gag atc att gag tct gcc tct cac ggg gaa tgc 4591 Arg Arg His Thr Arg Glu Ile Ile Glu Ser Ala Ser His Lily Glu Cys cca cgc att gac aac ctt gat aca ttg cag tgc aag ttg gca gac ata 4639 Pro Arg Ile Asp Asn Leu Asp Thr Leu Gln Cys Lye Leu Ala Asp I1e cta caa gac tca ggg aaa ttt ctg cta gtg ttg gat gat gtt tgg ttt 4687 Leu Gln Asp Ser Oily Lys She Leu Leu Val Leu Aap Asp Val Trp She gaa cct ggc agc gac sgg gaa tgg gac caa ctg ttg gca cca cta gtt 4735 Glu Pro Gly Ser Asp Arg Glu Trp Aap Gln Leu Leu Ala Pro Lau Val tcc caa cac cca gga agc aaa gtt ttg gta act tct gga cgg gat aca 4783 8er Gln His Pro Gly Ser Lya Val Leu Val Thr Sar Gly Arg Aap Thr ttt cca gtt get ctc tgc tgt caa gaa gtg cgt ttg ctg aaa aac cta 4831 Phe Pro Val Ala Leu Cys Cys Gln Glu Val Arg Leu Leu Lys Asn Leu gga gat get cag ttc ttg gca ctt ttc aaa cac cat gca ttc tct gga 4879 Gly Asp Ala Gln Phe Leu Ala Lau Phe Lys His His Ala Phe Ser Gly cca asc atc aga aat cca cag ttg tgt gca agg ctg gaa get ttt gca 4927 Pro Asa Ile Arg Asn Pro Gln Leu Cys Ala Arg Lau Glu Ala Phe Ala gag aag att get aaa agg ctt gga aaa tct cct ttg gca gca aaa gtt 4975 Glu Lya Ile Ala Lye Arg Leu Gly Lya Ser Pro Leu Ala Als Lys Val gtg ggt tcc caa ttg aaa gga aaa aca tgt atc act gca tgg aag gat 50x3 Val Gly Ser Gln Leu Lya Gly Lya Thr Cys Ila Thr Ala Trp Lya Aap get ctt act ata aag att gag saa tta agt gag ccc atg agc get ctg 5071 Ala Leu Thr Ile Lys Ile Glu Lye Leu Ser Glu Pro Met Ser Ala Leu ttg tgg agt tat gag aag tts aat eca tgt ttg cag agg tgc ttc cta 5119 Leu Trp Ser Tyr Glu Lys Leu Aan Pro Cys Leu Gln Arg Cys Phe Leu tst tgc agc ttg ttt cca aaa ggc cac aag tat tta att ggt gag ttg 5167 Tyr Cys Ser Leu She Pro Lya Gly Hia Lya Tyr Leu Ila Gly Glu Leu gtt cat ctt tgg atg gca gag ggc ctc att gat tcg tgc aac caa aac 5215 Val His Leu Trp Met Ala Glu Gly Leu Ile Aap Ser Cya Aen Gln Asn WO 99/45118 PC"T/AU99/00130 aag aga gtg gaa gat att ggg agg gac tgc ttc aag gag atg atc tct 5x63 Lys Arg Val Glu Asp Ile Gly Arg Aep Care Phe Lys Glu Met Ile Ser gtt tcg ttc ttt cas aaa ttt ggt aag gaa aag gaa cac act ccc aca 5311 Val Ser Phe Phe Gln Lys Phe Gly Lye Glu Lye Glu His Thr Pro Thr tac tat gtt atg cat gat ctt ctt cat gat ctg gca gaa tca ctc tcc 5359 Tyr Tyr Val Met His Asp Leu Leu His Asp Leu Ala Glu Ser Leu Ser 5a0 525 530 aaa gag gtc tac ttc aga ttg gaa gac gat aat gtg aca gaa ata ccg 5407 Lys Glu Val Tyr Phe Arg Leu Glu Asp Asp Asa Val Thr Glu Ile Pro tcc act gtt cga cat cta tca gtt cgt gtt gag agt atg aag cgg cac 5455 Ser Thr Val Arg His Leu Ser Val Arg Val Glu Ser Met Lys Arg His aag aat agt atc tgc aag cta tat cat tta cgc act att atc tgc atc 5503 Lys Asn Ser Ile Cys Lys Leu Tyr His Leu Arg Thr Ile Ile Cys Ile gac ccc cta atg gat gat gta sat gac att ttt aat cag gta cta cag 5551 Asp Pro Leu Met Asp Asp Val Asn Asp Ile Phe Asa Gln Val Leu Gln tat ttg aag aag ttg cgc gta cta tat ttg tca tct tat aac agt agt 5599 Tyr Leu Lys Lys Leu Arg Val Leu Tyr Leu Ser Ser Tyr Asn Ser Ser aag ttg cca gaa tct gtt ggt gag ttg aag cac ctt cgg tat ttg aac 5647 Lys Leu Pro Glu Ser Val Gly Glu Leu Lya His Leu Arg Tyr Leu Asn atc atc agc aca ctg att tct gaa tta cca aga tca ttg tgt acc ctt 5695 Ile Ile Ser Thr Leu Ile Ser Glu Leu Pro Arg Ser Leu Cys Thr Leu tac cac ttg cag cta ctt ctg tta aat gac aaa gtt gag agt ttt cct 5743 Tyr His Leu Gln Leu Leu Leu Leu Asn Asp Lys Val Glu Ser Phe Pro gaa aaa ctt tgt aat tta tgg aaa tta cgt cat ctt gaa cgg cac caa 5791 Glu Lys Leu Cys Asn Leu Trp Lys Leu Arg Hie Leu Glu Arg His Gln gat tgg gat cat gat sta sta ttc cca tat aaa gaa gca cca cat caa 5839 Asp Trp Asp His Asp Its Ile Phe Pro Tyr Lys Glu Ala Pro Hie Gln att cct aac att ggc aag tta act tca ctt caa caa ttt gag gaa ttt 5887 Ile pro Asn Its Gly Lys Leu Thr Ser Lau Gln Gln Phe Glu Glu Phe tct gtg caa aag saa aaa gga tat gag ttg caa caa ctg agg gac atg 5935 Ser Val Gln Lys Lys Lys Gly Tyr Glu Leu Gln Gln Leu Arg Asp ldet aat gag att cga ggc tgt tta cgt ctc aca aat ctt gag aat gtc act 5983 Asn Glu Ile Arg Gly Cys Leu Arg Lsu Thr Asa Lsu Glu Asn Val Thr ggc aag gat caa gcc ttt gaa tcg aag ctg cat cac aaa att cat ctt 6031 Gly Lys Asp Gln Ala Phe Glu 8sr Lys Leu His His Lys Ile His Leu gac aca ttg cat ctt gtt tgg agg tgc asa aat gac acc aat gca gag 6079 Asp Thr Lsu His Leu Val Trp Arg Cys Lys Asn Aep Thr Asn Ala Glu gat agt tta cat ttg gag att cca ccg cct caa ctt ggg gat ctt aca 6127 Aep 8er Leu His Leu Glu Its Pro Pro Pro Gln Leu Gly Asp Leu Thr att aac ggt tac aaa tct tca aaa tat ccc gac tgg tta ctt gat get 6175 Its Asn Gly Tyr Lys Ser Ser Lys Tyr Pro Asp Trp Leu Leu Asp Ala tca tat ttt gag ttt ttg aaa tct tta aga ttt gtt aat tgc act gca 6223 Ser Tyr Phe Glu She Leu Lys Ser Leu Arg Phe Val Asn Cys Thr Ala tta caa agc ata cca tcc aac acc gaa ctg ttt agg aat tgc tct tca 6271 Lsu Gln 8er Ile Pro Ser Asn Thr Glu Lsu Phs Arg Asn Cys 8er Ser ctt gtc ctc tgg aat gtg cca aac ctg aaa aca tta cet tgt ctt cca 6319 Leu Val Leu Trp Asn Val Pro Asn Leu Lye Thr Lsu pro Cys Leu Pro cca ggc ctt caa aag tta gaa att gta gaa tgc cca ctg ctt ata ttt 6367 Pro Gly Leu Gln Lys Leu Glu Ile Val Glu Cys 9ro Leu Leu Ile Phs WO 99/4511$ PCT/AU99/00130 att tca aat gat gag ctg gaa cat caa gag cag aga gag aat atc acg 6415 Ile Ser Asn Asp Glu Leu Glu His Gln Glu Gln Arg Glu Asn Ile Thr agt ata gac cac ttg gta tca cag ctt agt tta atc tgg gag gta gat 6463 Ser Ile Asp His Leu Val Ser Gln Leu Ser Leu Ile Trp Glu Val Asp tct gga tca cat att agg agt aca ctc tca ttg gaa ctt tca ttt ctg 6511 Ser Gly Ser His Ile Arg 8er Thr Leu Ssr Leu Glu Leu Ser Phe Leu aag cag ctg atg ata tcg atg cat get gat atg tcg cat gtt caa aat 6559 Lys Gln Leu Met Ile Ser met His Ala Asp Met Ser His Val Gln Asn ctt gaa att tct cta gaa ggt gaa aaa gat gaa gca tcg gca gaa gag 6607 Leu Glu Ile Ser Leu Glu Gly Glu Lys Asp Glu Ala Ser Ala Glu Glu gat atc atc aag gca tgg sca tac tgt cac gag caa agg atg gga gtc 6655 Asp Ile Ile Lys Ala Trp Thr Tyr Cys His Glu Gln Arg Met Gly Val aca tat gga agg agc atg gtg ctg ccg ctg gtt cca cca tca gga ctt 6703 Thr Tyr Gly Arg Ser Met Val Leu Pro Leu Val Pro Pro Ser Gly Leu tgt gag ctt ttt ctt tct tca tgc agt att aca gat gga get tta get 6751 Cys Glu Leu Phe Leu Ser Ser Cps Ser ile Thr Asp Gly Ala Leu Ala gtt tgc ctt gat ggc ctg get tca ctg aaa aga ttg gtc ttg gta gat 6799 Val Cys Leu Asp Gly Leu Ala Ser Leu Lys Arg Leu Val Leu Val Asp att atg act tta act gca ctt cct tca gaa gag atc ctc caa cat ttg 6847 Ile Met Thr Leu Thr Ala Leu Pro Ser Glu Glu Ile Leu Gln His Leu 1015 10x0 10x5 aaa aaa ctt gac cac ttg tac atc tgg cgt tgc tgg tgt ctc aag tca 6895 Lys Lys Leu Asp His Leu Tyr Ile Trp Arg Gars Trp Gars Leu Lys Ser tta ggg ggc tta cga get get acc tca ctt tca act gtt aga ttg gtt 6943 Leu Gly Gly Leu Arg Ala Ala Thr Ser Lau Ser Thr Val Arg Leu Val tcc tgc cct tct tta gag ttg gca cgt gga gca gaa tgt ttg cca tta 6991 Ser Cys Pro Ser Leu Glu Leu Ala Arg Gly Ala Glu Cys Lsu Pro Leu tec ctt aag acg ctc gtt ata gtc agt tgc gtg ctt gca get gac ttc 7039 Ser Leu Lys Thr Leu Val Ile Val 8er Cys Val Leu Ala Ala Asp Phe ctc tgt act gag tgg cca tac att gat aca ata agc ata acc aat tgc 7087 Leu Cps Thr Glu Trp Pro Tyr Ile Aep Thr Ile Ser Ile Thr Asn Gds aga agc acc aga tgc ttg tcc gtt ggt agt ctc acc tct gtt aaa tca 7135 Arg Ser Thr Arg Cys Leu Ser Val Gly Ser Leu Thr Ser Val Lys Ser lllo llls llao ttc ata ctg gag cac ttg cca gat tta tgt acg ctc gaa gga ttg tct 7183 Phe Ile Leu Glu His Leu Pro Asp Leu Gys Thr Leu Glu Gly Leu Ser tcc ctg caa ctg cac aac gtg cat ttg att get att ccg aaa ctc gtc 7231 Ser Leu Gln Leu His Asn Val His Leu Ile Ala Ile Pro Lys Leu Val cct gag tgt atc tca cag ttt aga gtc cag aga tca ctc tat att ggc 7x79 Pro Glu Cys Ile Ser Gln Phe Arg Val Gln Arg Ser Leu Tyr Ile Gly agt ctc gtg ata ctc aac gag atg ctc tca get gaa ggt ttc aca ctt 73x7 Ser Leu Val Ile Leu Aan Glu Met Leu Ser Ala Glu Gly Phe Thr Lau cca gca ttt ctc tgt ctt caa gga tgc aag gaa cca ttt gtt tca ttt 7375 Pro Ala Phe Leu GSra Leu Gln Gly Cys Lys Glu Pro Phe Val Ser Phe gaa gaa tat gca aat ttt aca tcc gtc cgg aga ctg aga tta tct gga 7423 Glu Glu Ser Ala Asn Phe Thr Ser Val Arg Arg Leu Arg Leu Ser Gly 1205 1x10 lal5 tgt caa atg act tcc ctt cca aaa aat cta gag cgc ttc tcc aat ctg 7471 Cys Gln Met Thr Ser Leu Pro Lye Asn Lsu Glu Arg Phe Ser Asn Leu 1x20 1225 1x30 1235 aag gtg atc gac att gtt agg tgc ccc aac ata tca tct tta cca gat 7519 Lys Val Ile Asp Ile Val Arg Cys Pro Asn Ile Ser 8er Leu Pro Asp WO 99/4511$ PCT/AU99/00130 ctg cca tcc tcc ctc ctt cag ata tgc ata tgg gat gat tgc gag cgc 7567 Leu Pro Ser Ser Leu Leu Gln Ile Cys Ile Trp Aep Aap Gyre Glu Arg 1x55 1x60 1265 ttg aag gag sgt tgt cga gca cct gat gga gaa agt tgg cca aag att 7615 Leu Lys Glu Ser Cys Arg Ala Pro Asp Gly Glu Ser Trp Pro Lys Ile gca cat att cgc agg aag tac ttt tta aaa gtg tgatttagat ccctacatac 7668 Ala Hia Ile Arg Arg Lys Tyr Phe Leu Lys Val aaataaacgg aaaggtsaaa actactgtca gccacctttg cactgtcaaa gatatatagc 77x8 cctttgtaac attagattct gttattttgc cttttaaaat actccattta cttcaaaatt 7788 ccattaattt gtttacttca aattgatagc aggcccaaga tgtgtgccct ccaggaacct 7848 ggtacgagcc ggagtgctgc tctcaggctg gcaactttta cagttcaact atctgtccta 7908 tgattacctc actggccggt gttgatccct tcggttggtc tcctgctttg ctgctgattg 7968 cttcatgcat tgcctctact ccggcacctc ttgtcctcta gtgctacaat ccatgatggt 8028 ggattttgta ttcctgtttt cggctatctt ctccatgctt caccaaccga acaattaccc 8088 gaagtagttt ctcatttaca tgatgctttg tcatgctgtc stgttacctt ctcctgaagg 8148 atgtcgtact tttgatttgt gcggattgtt attaggatcc gttgacctgc aggtcgac 8206 <210> 6 <211> 1294 <al2> PRT
<213> Hordeum ap.
<400> 6 Met Ala Glu Val Ala Leu Ala Gly Ala Ala Leu Ser Val Ser Leu Asn Leu Ala Ala 8er Pro Val Leu Lys Lys Leu Leu Ala Aan Ala Ser Ile ZO a5 30 Tyr Leu Gly Val Aep Met Ala Lau Glu Leu Leu Glu Leu Glu Thr Thr Ile Leu Pro Gln Phe Glu Lau Val Ile Glu Ala Ala Asn Lys Gly Asn His Arg Pro Lys Leu Asp Lys Trp Leu Gln Glu Leu Lys Glu Gly Phe Tyr Leu Ala Glu Asp Leu Leu Asp Asp His Glu Tyr Asn Lsu Leu Lys Arg Gln Ala Lys Oly Lys Asp pro Leu Pro Ala Asn Gly Ser Ser Ile Ser Asn Thr Phs Met Lys Pro Leu Arg Ala Ala Ser Ser Arg Leu Ser Asn Phe Ser Ser Glu Asn Arg Lys Leu Ile Gln Gln Leu Asn Gly Leu Lys Ala Thr Leu Ala Lys Ala Lys Asp Phe Arg Glu Leu Leu Phe Leu Pro Ser Gly Tyr Ser Ala Glu Gly Sar Thr Ile Pro Ser Ala Val Val Pro Glu Thr Ser Ser Ile Ala Pro Leu Lys VaI Ile Gly Arg Arg Lys Asp Arg Asn His Ile Ile Asn Cys Leu Thr Lys Thr Thr Val Thr Thr 195 a00 205 Lys Ser Ser Lys Thr Met Tyr Ser Gly Leu Ala Ile Val Gly Ala Gly a10 Z15 ZZO
Gly Ile Gly Lys Ser 8er Leu Ala Gln Leu Val Tyr Asn Ser Lys Arg 225 230 a35 a40 Val Lys Lye His Phe Asn Val Arg Met Trp Ile Ser Ile Ser Arg Lys Z45 a50 255 Leu Asp Val Arg Arg His Thr Arg Glu Ile Ile Glu Ser Ala Ser His 260 a65 270 Gly Glu Cys Pro Arg Ile Asp Asa Leu Asp Thr Leu Gla Cys Lys Leu Ala Asp Ile Leu Gln Aap Ser Gly Lys Phe Leu Leu Val Leu Asp Asp 290 a95 300 WO 99/4511$ PCT/AU99100130 Val Trp She Glu Pro Gly Ser Asp Arg alu Trp Asp Gln Lau Leu Ala 305 310 315 3a0 Pro Leu Val Ser Gln His Pro Gly Ser Lys Val Leu Val Thr Ser Gly Arg Asp Thr Phe Pro Val Ala Leu Cys Cys Gln Glu Val Arg Leu Leu Lys Aen Leu Oly Asp Ala Gln Phs Leu Ala Leu Phe Lys His His Ala Bhe Ser Gly Pro Asn Ile Arg Asn Pro Gln Leu Cys Ala Arg Leu Glu Ala Phe Ala Glu Lys Iie Ala Lys Arg Leu Gly Lya Ser Pro Leu Ala Ala Lys Val Val Gly Ser Gln Lau Lys Gly Lye Thr Cys Ile Thr Ala Trp Lys Asp Ala Leu Thr Ile Lys Ile Glu Lys Leu Ser Glu Pro Met Ser Ala Leu Leu Trp Ser Tyr Glu Lye Leu Asn Pro Cys Leu Gla Arg Cys Phe Leu Tyr Cys Ser Leu Phe Pro Lys Gly His Lys Tyr Leu Ile Gly Glu Leu Val His Leu Trp Met Ala Glu Gly Leu Ile Asp Ser GSrs Asn Gln Asn Lys Arg Val Glu Asp Ile Gly Arg Asp Cys Phe Lys Glu Mat Ile Ser Val Ser Phe Phe Gln Lys Phe Gly Lys Glu Lys Olu His Thr Pro Thr Tyr Tyr Val Met His Asp Leu Leu His Asp Leu Ala Glu 515 520 5a5 Ser Leu Ser Lys Glu Val Tyr Phe Arg Leu Glu Asp Asp Asn Val Thr Glu Ile Pro Ser Thr Val Arg His Leu Sar Val Arg Val alu Ser Met Lys Arg His Lys Asa Ser Ile Cys Lys Leu Tyr His Leu Arg Thr Ile Ile Cys Ile Asp Pro Leu Met Asp Asp Yal Asn Asp Ile Phe Asa Gln Val Leu Gln Tyr Leu Lys Lya Leu Arg Val Leu Tyr Leu Ssr Ser Tyr Asn Ser Ser Lys Leu Pro Glu Ser Val Gly Glu Leu Lya 81s Leu Arg Tyr Leu Asn Ile Ile Ser Thr Leu Ile Ser Glu Leu Pro Arg Ser Leu Cys Thr Leu Tyr His Leu Gln Leu Leu Leu Leu Asn Asp Lys Val Glu Ser Phe Pro Glu Lys Leu Cys Asa Leu Trp Lys Leu Arg His Leu Glu Arg His Gln Asp Trp Asp His Asp Ile Ile Phe Pro Tyr Lys Glu Ala Pro His Gln Ile Pro Asa Ile Gly Lya Leu Thr Ser Leu Gln Gla phe Glu Glu Phe Ser Val Gla Lys Lys Lys Gly Tyr Glu Leu Gla Gln Leu Arg Asp Met Asn Glu Ile Arg Gly Gars Leu Arg Leu Thr Asn Leu Glu Asn Val Thr Gly Lys Asp Gla Ala Phe Glu Ser Lys Leu His His Lys Ile His Leu Asp Thr Leu His Leu Val Trp Arg Cars Lys Asa Asp Thr Asa Ala Glu Asp Ser Leu His Leu Glu Ile Pro Pro 8ro Gln Leu Gly Asp Leu Thr Ile Asn Gly Tyr Lys Ser Ser Lys Tyr Pro Asp Trp Leu Leu Asp Ala Ser Tyr Phe Glu Phe Leu Lys Ser Leu Arg Phe val Asn Gyre Thr Ala Leu Gln Ssr Ile Pro Ser Asn Thr Glu Leu Phe Arg Asn 820 8a5 830 Cys Ser 8er Leu Val Leu Trp Aan Val Fro Asn Lsu Lys Thr Leu Pro Gars Leu Pro Pro Gly Leu Gln Lys Leu Glu Ile Val Glu Cys Pro Lwu Leu Ile Phe Ile 8er Asn Asp Glu Leu Glu His Gla Glu Gln Arg Glu Asn Ile Thr Ser Ile Asp His Leu Val Ssr Gln Leu Ser Leu Ila Trp Glu Val Asp Ser Gly 8er His Ile Arg 8er Thr Leu 8er Lsu Glu Leu 8er Phe Leu Lys Gln Leu Met Ile 8er Met His Ala Asp Met 8er His Val Gln Asn Leu Glu Ile 8er Leu Glu Gly Glu Lys Asp Glu Ala 8sr Ala Glu Glu Asp Ile Ile Lys Ala Trp Thr Tyr Cys His Glu Gln Arg Met Gly Val Thr Tyr Gly Arg Ser Met Val Leu Pro Lsu Val Pro Pro 8er Gly Lsu Cys Glu Leu Pha Lsu Ser 8er Cars Bar Ile Thr Asp Gly Ala Lau Ala Val Cys Lau Asp Gly Leu Ala 8er Lsu Lys Arg Leu Val Leu Val Asp Ile Met Thr Lsu Thr Ala Leu Pro Ssr Glu Glu Ile Leu Gln His Leu Lys Lys Leu Asp His Leu Tyr Ile Trp Arg Cys Trp Gars Oa5 1030 1035 1040 Leu Lys Sar Leu Gly Gly Leu Arg Ala Ala Thr Ser Lsu 8er Thr Val Arg Leu Val 8er Cya Pro Ser Leu Glu Leu Ala Arg Gly Ala Glu Cys _ Lsu Pro Leu Ser Leu Lys Thr Leu Val Ile Val Ser Cys Val Leu Ala Ala Asp Phe Leu Cys Thr Glu Trp Pro Tyr Ile Asp Thr Ile Ser Ile Thr Asa Cars Arg Ser Thr Arg Cys Leu Sar Val Gly Ser Leu Thr Ssr 105 1110 1115 11a0 Val Lys Sar Phe Ile Leu Glu His Leu Pro Asp Leu Cys Thr Leu Glu Gly Leu Bar Ser Leu Gla Leu His Asa Val His Leu Ile Ala Ile Pro Lys Leu Val Pro Glu Cys Its Ser Gla Phe Arg Val Gln Arg Ser Leu Tyr Its Gly Ser Leu Val Ile Leu Asa Glu Met Leu Ser Ala Glu Gly Phe Thr Lau Pro Ala Phe Lau Cys Leu Gla Gly Cys Lys Glu Pro Phe 185 1190 1195 1x00 Val Ser Phe Glu Glu Ser Ala Asn Phe Thr Ser Val Arg Arg Leu Arg 1a05 1a10 1a15 Leu Ser Gly Cys Gla Met Thr Ser Leu Pro Lys Asa Leu Glu Arg Phe 1x20 laa5 1x30 Ser Asn Leu Lys Val Ile Asp Its Val Arg Cys Pro Asa Ile Ser Ser 1x35 1a40 1a45 Lsu Pro Asp Leu Pro Ssr Ser Leu Lsu Gln Ile Gds Ile Trp Asp Asp la5o lass laso ors Glu Arg Leu Lys Glu Ssr Cys Arg Ala Pro ASp Gly Glu Ser Trp 265 1a90 1a75 1a80 Pro Lys Ile Ala His Ile Arg Arg Lys Tyr Phe Leu Lys Val 1a85 1a90 <a10> 7 <all> 5763 <ala> DNA
<a13> Hordeum sp.
4a wo 99iasiis Pcr~AV99rooi3a _ <azo>
<221> CDS
<aaz> (715)..(4551) <400> 7 ggtaaaaagg gcaagtagcc ggatcagatc tgttattgcc ttgtttgacc tgctgtttgg 60 ggattttagc ctcctctcat tattagctcc tactatctgc ttctcctgca accttgcttc 120 agcgctctgt ttctcactcc ttcatctgtg tacgatcacc actatgctcc tctgttttcc 180 tctgcaaatc tggatgtagg gttctgttta ttttctctgt tgctgcaacc ggctaattta X40 acccgtgatt tcctactcca acaggttttc tatcaactgc tgtattcatt ggttgtttct 300 actgcctctg atgtgaagca ctctcattgg tcatctatca tttattctgg taatttccca 360 cactcaaaag tagtattctt ggtttttttt acgttgccta cggatctgcc agtagatttc 4Z0 gtatctccat taaagttgac aaatttatct cctcatttcc cactcaacat gtttttcatc 480 catcactgcg tccttccagt gctcaccttt cgatttattc acgacattcc acacctaaaa 540 aaatcaagga aagagaggaa aattgtcttt atagcttttg ctcctatttt tcattgtact 600 tgttcatctc catacaatgt gtttcatcat tttgtatctt tatatggatc gagtttgttt 660 tcttattttc ttctataaac tcgctttcag cctagtagcc tcacctacag tccg atg 717 Met gca gaa gtg gcc tta get ata get gcc tta aga ttg get gcg ttg ccg 765 Ala Qlu Val Ala Leu Ala =le Ala Ala Leu Arg Leu Ala Ala Leu Pro gtc ttg aag aag ctc cat get aat get tca acg tac ctt gga gtg aac 813 Val Leu Lys Lys Leu His Ala Asn Ala Ser Thr Tyr Leu Qly Val Asn 20 a5 30 atg gcg cgt gag atc cat gaa ctg gag acc act stc atg cca cag ttt 861 Met Ala Arg Qlu I1. His G~lu Leu Cilu Thr Thr Ile Met Pro aln Phe gaa cta gtg att gaa gca gca gac aag gga aac cac agg ccc aag ttg 909 alu Leu Val Ile alu Ala Ala Asp Lys Qly Asn His Arg Pro Lys Leu gac aaa tgg ctt cag gaa ctc aaa gaa agc ttc tac ctg get gaa gac 957 Asp Lys Trp Leu Gln Glu Leu Lys Glu Ser Bhe Tyr Leu Ala Glu Asp ttg cta gac gag cat gag tac aac atc ctc aag cac aaa gca aag ggt 1005 Leu Leu Asp Glu His Glu Tyr Asn Ile Leu Lye His Lys Ala Lys Gly aag gat tcc atg ccg gcc aat ggt tcc tcc atc agc aac act ttt atg 1053 Lya Asp Ser Met Pro Ala Aan Gly Ser Ser Ile Ser Asn Thr Phe Mst aag cct ctg cgt tct gca tca agc agg ctg tcc aat ctg agt tca gag 1101 Lys Pro Leu Arg Ser Ala Ser Ser Arg Lau Ser Asn Leu Ser Sar Glu aac aga aag cta gtt cgc cat ctt aag gaa ctg aag gcc acc ctc gcg 1149 Asn Arg Lys Leu Val Arg Hie Leu Lya Glu Leu Lya Ala Thr Leu Ala aaa gcc aag gac ttc cat cag ctt ctc tgt tta cct get ggt cat aac 1197 Lys Ala Lye Asp Phe His Gln Leu Leu Cps Leu Pro Ala Gly His Asn gca gag aga ccc gcc ata cca tca gat gtt gtt cct gag atc aca tcg 1x45 Ala Glu Arg Pro Ala Ile Pro Ser Asp Val Val Pro Glu Ile Thr Ser cta cca cct atg aaa gtg att ggt cgt gac aag gat cge gat cat ata 1293 Leu Pro Pro Met Lys Val Its Gly Arg Asp Lys Asp Arg Asp His Ile ata gag tgt ett acc aag gta act get act acc gag tct agt aca act 1341 Ile Glu Cys Leu Thr Lys Val Thr Ala Thr Thr Glu Ser Ser Thr Thr atg tac tcg ggt ttg gcc att gtt gga gtt gga ggc atg gga aag tcc 1389 Met Tyr Ser Gly Leu Ala Ile Val Gly Val Gly Gly Met Gly Lys Ser a10 215 Za0 aa5 acc ttg get cag ctt gtt tac sat gac aag agg gta aaa gaa tat ttt 1437 Thr Leu Ala Gln Lau Val Tyr Asn Asp Lys Arg Val Lys Glu Tyr Phe Z30 a35 a40 gat gtg aca atg tgg gta agc atc tca cgt aaa ctc gat gtc cgc cgt 1485 Asp Val Thr Mat Trp Val Ser Ila Ser Arg Lys Leu Asp Val Arg Arg cat aca cgg gag atc atc gag tct gcc tct aag ggg gaa tgc cca cgc 1533 His Thr Arg Glu Ile Ile Glu Ser Ala Ser Lys Oily Glu Cars Pro Arg Z60 265 a70 att gat aac ctt gat act ctg cag tgc aag ttg aca gac ata ctg caa 1581 Ile Asp Asn Leu Asp Thr Leu Gln Cys Lys Leu Thr Asp Its Leu Gln 275 280 aB5 gaa tca ggg aaa ttc ctg ctt gtg ctg gat gac gtt tgg ttt gaa ctt 1629 Glu Ser Gly Lys Bhe Leu Leu Val Leu Asp Asp Val Trp Phe Glu Leu a90 a95 300 305 ggc agt gaa agg gaa tgg gac csa ctg tta gcc cct cta gtt tcc aga 1677 Gly 8er Glu Arg Glu Trp Asp Gln Leu Leu Ala Bro Leu Val Ser Arg cag acg gga agc aaa gtt ttg gta act tct cga cgg gat aca ttt cca 1725 Gln Thr Gly Ser Lys Val Leu Val Thr Ser Arg Arg Asp Thr Phe Pro 3a5 330 335 get act ctt tgc tgt gaa gtg tgt cct cta gag aag atg gat gat get 1773 Ala Thr Leu Cys Cys Glu Val Cys Pro Leu Glu Lys Met Aep Asp Ala cag ttc ttg gca ctc ttc aaa cat cat gca ttc tct gga cca gaa atc 1821 Gln Phe Leu Ala Leu Bhe Lys His His Ala Phe Ser Gly Pro Glu Ile aga aat ccg cag ttg cgt gaa aag ttg gaa gag ttt tca sag aag att 1869 Arg Asn Pro Gln Leu Arg Glu Lye Leu Glu Glu Phe Ser Lys Lys Ile get aaa cgg ctt gga caa tct cct ttg gcg gca aaa gtt atg ggt tcg 1917 Ala Lys Arg Leu Gly Gln Ser Pro Leu Ala Ala Lys Val Met Gly Ser cag ttg aaa ggg aaa aca gat atc act gca tgg aag gat get cta act 1965 Gln Leu Lys Gly Lys Thr Asp Ile Thr Ala Trp Lys Asp Ala Leu Thr atg aag att gac asa cta agc gat ccc atg aga get ctg ttg tgg agt x013 Met Lys Ile Asp Lye Leu Ser Asp Pro Met Arg Ala Leu Leu Trp Ser 420 4a5 430 tat gag aaa tta gat cca tgt ctg cag aga tgc ttt tta tat tgc agc 2061 Tyr Glu Lya Leu Asp Pre Cys Leu Gln Arg Cys phe Leu Tyr ors Ser ttg ttt ccg aaa ggc cac aag tat gtc att gat gat ttg gtt cat ctt 2109 WO 9914511$ PC'TIAU99100130 Lsu Phe Pro Lys Gly His Lys Tyr Val Ile Asp Asp Lsu Val His Lsu tgg atg gca gag gga ctt gtt gat tca tgc aac caa aac aag aga gta 2157 Trp Met Ala Glu Gly Leu Val Asp Ser Cys Asn Gln Asn Lys Arg Val gaa gtt gtt ggg agg gat tgt ttc cat gaa atg ata tct gtt tag ttc aao5 Glu Val Val Gly Arg Aap Cys Phs His Glu Met Its Ssr Val Ssr Phs ttt caa cca gtt gat gag aaa tac act gac acg tac tac gtt atg cat ZZ53 Phe Gln Pro Val Asp Glu Lys Tyr Thr Asp Thr Tyr Tyr Val Met His gat ctc ctt cat gat atg gaa gaa tca ctc tcc aaa gaa gac tac ttc x301 Asp Lsu Lsu His Asp Leu Ala Glu Sar Leu Ser Lys Glu Asp Tyr Phe 515 5Z0 5a5 aga ttg gaa gat gat aag gtg aca gaa ata cca tcc act gtt cga aat x349 Arg Leu Glu Asp Asp Lys Val Thr Glu Ile Pro Ser Thr Val Arg His ata tct gtt cgt gtt gaa agt atg aca cag cac aag caa agt att tgc 2397 Leu Ser Val Arg Val Glu Ser Met Thr Gln His Lys Gln Ser Ile Cys aag cta cat cat tta cgc act att atc tgc ata gaa ccc cta gta gat x445 Lys L.u His His Lsu Arg Thr Ile Its Gars Ile Asp Pro Lau Val Asp gat gtt agt gac ctt ttt aat cag ata cta cag aat ttg aag aag tta X493 Asp Val Ser Asp Lau Phs Asn Gln Ile Leu Gln Asn Lsu Lys Lys Leu cgc gta cta tat ttg tca tca taa agt agc agt cag ttg cca gaa tat 2541 Arg Val Leu Tyr Leu Ser Ser Tyr Sar Ser Ser Gln Lau Pro Glu Ser gtt ggt gag ttg aag cat ctt cgg tat ttg sac atc atc ggg aca atg 2589 Val Gly Glu Leu Lys His Lsu Arg Tyr Leu Asn Ile Ile Gly Thr Leu att tct gaa ttg cca aga tca ctg tgt act ctt tac cac ttg cag tca 2637 Its Ser Glu Leu Pro Arg Ser Leu Cys Thr Lau Tyr His Lsu Gln Ser ctt ctg tta aat gac agt gtg aag agt ttg cct gaa aac atc tgc aat 2685 Leu Leu Leu Asn Asp 9er Val Lys Ser Leu Pro Glu Asn Ile Gds Asn tta agg aag tta cgg cat ctt gaa cgg gat gag ttg gca ctg cct aag 2733 Leu Arg Lys Leu Arg His Leu Glu Arg Asp Glu Leu Ala Leu Pro Gla att cct aac att ggc aag cta act ttg ctt caa caa ttg gat aaa ttt 2781 Ila Pro Asn Ile Gly Lys Leu Thr Leu Leu Gln Gln Lau Asp Lys Phe tct gtg cag aag aaa aag gga ttt gag ttg gas cag ctg agg gac atg Z8a9 Ser Val Gln Lys Lye Lys Gly Phe Glu Leu Glu Gln Leu Arg Asp Met aac gag att cgt ggc cac tta agt gtc gaa cat ctt gag aat gtc act 2877 Asn Glu Ile Arg Gly His Leu Ser Val Glu His Leu Glu Asn Val Thr gga aag gat caa gcc ata gaa tcg aag cta tat cag aaa agt cat ctt a9a5 Gly Lya Asp Gln Ala Ile Glu Ser Lys Leu Tyr Gln Lys Ser His Leu gac agc ttg cac ctt ggc tgg aat ctc gga aat aac acg act gca gag 2973 Asp 8er Leu His Leu Gly Trp Asn Lau Gly Asn Asa Thr Thr Ala Glu gat agc tta cat ttg gag att cta gaa ggc ctg acc cca ccg cct caa 3021 Asp Ser Leu His Leu Glu Ile Leu Glu Gly Leu Thr Pro Pro Pro Gln att tcg gcc ctt tca att gag ggt tac gaa tct tgg aaa tat cca ggc 3069 Ile Ser Ala Leu Ser Ile Glu Gly Tyr Glu Ser Trp Lys Tyr Pro Gly tgg tta att gat ggt tcg tat ttt gag aat ttg aat tat ttg aga ttt 3117 Trp Leu Ile Asp Gly Ser Tyr Phe Glu Asn Leu Asn Tyr Leu Arg Phs ttt ggt tgc aga aaa tta caa atc tta cca tcc aat act gag ctg ttt 3165 Phe Gly Cys Arg Lys Leu Gln Ile Leu Pro Ser Asn Thr Glu Leu Phe gtg aat tgc act tca ctt ctc ctc cag ggt ttg tca aac ctg aac aca 313 Val Asa Cys Thr Ser Leu Leu Leu Gln Gly Leu Ser Asn Leu Asn Thr 8a0 825 830 tta cct tgt ctt cca cta ggc ctt aag gtg tta aaa gtg cag cgc tgc 3261 WO 99/45118 PC'TIAU99/00130 Leu Pro Cys Leu Pro Leu Gly Leu Lys Val Leu Lys Val Gln Arg Cps cca ctg ctc ata ttt att tcc cat gat gag ctg gsa cat aat gac cag 3309 Pro Leu Leu Ile Phe Ile 8er His Asp Glu Leu Glu His Asn Asp Gln aga gag aat agc aca agg acc sac cat ttg gca tca caa ctt ggt ttg 3357 Arg Glu Asn Ser Thr Arg Thr Asn His Leu Ala Ser Gla Leu Gly Leu atg tgg gag gtg gat tca gga tca ggt att agc act gta ctc tta tcg 3405 Met Trp Glu Val Asp Ser Gly Ser Gly Ile Ser Thr Val Leu Leu Ser gaa tgt tca ttt ctg aag cag ctg atg ata ttc aca cat get gat atg 3453 Glu ire Ser Phe Leu Lys Gln Leu Met Ile Phe Thr Hia Ala Asp Met tca cat gtt caa aac ttt gaa agt get cta cag aga gcg aaa aat gga 3501 Ser His Val Gln Asn phe Glu Ser Ala Leu Gln Arg Ala Lys Asa Gly gtt ttg gta aaa gag gat atc atc aag gca tgg ata tgc tgt cat gag 3549 Val Leu Val Lys Glu Asp Ile Ile Lys Ala Trp Ile Cars Cys His Glu caa agc atg aga ctc atg tat gaa cgg agg stt ggg ctg cca ttg gtt 3597 Gln Ser Met Arg Leu Met Tyr Glu Arg Arg Ile Gly Leu Pro Leu Val cca cca tca ggc ctt cat gaa ctt cat ctt tct tca tgc agt att aca 3645 Pro Pro Ser Gly Lau His Glu Leu His Leu Ser Ser Gars Ser Its Thr gat gaa gca tta get gtt tgc ctt gat ggc ctg get toa ctg ggg agt 3693 Asp ~Glu Ala Leu Ala Val Cys Leu Asp Gly Leu Ala Ser Leu Gly Ser ttg ttc tta gaa aag att ttt aat tta act aaa ctt cct tca gaa gag 3941 Leu Phe Leu Glu Lys Ile Phe Asn Leu Thr Lys Leu Pro Sar Glu Glu gtc ctc aaa cat ctg gca aaa ctt ggc cac ttg aac atc aca gat tgc 3789 Val Leu Lys His Leu Ala Lys Lau Gly His Lau Asn Ila Thr Asp Cys 1010 1015 1020 lOZ5 tgg tgt ctt agg tca tta ggg ggc tta cga get get acc tct ctt gca 3837 Trp Cys Leu Arg Ser Leu Gly Gly Leu Arg Ala Ala Thr Ser Leu Ala cat ttt aca ttg aga tct tgc cct tct tta gag ttg gca cat gga gca 3885 His Phe Thr Leu Arg Ser Cys Pro Ser Leu Glu Leu Ala His Gly Ala gaa tgc ttg cca tta tcc att gag agc ctc tgg sta gaa aag tgc atg 3933 Glu Gys Leu Pro Leu Ser Ile Glu Ser Leu Trp Ile Glu Lys Cys Diet ctt gaa ggt aac ttc ctc tgt act gac tgg cca cac atg gat aaa att 3981 Leu Glu Gly Asn Phe Leu Gars Thr Asp Trp Pro His Met Aap Lys Ile tcc ata tgg aat tgc aga agc acg gca tgc ctg tct gtt ggt agt ctc 4029 Ser Ile Trp Asn Cys Arg Ser Thr Ala Cys Leu Ser Val Gly Ser Leu acc tct gtt aaa aaa tta tca ctg gat cgg ttg cca gat tta tgt atg 4077 Thr Ser Val Lys Lys Leu Ser Leu Asp Arg Leu Pro Asp Leu Cys Met ctc gag gga ttg tgt ttc ctg caa ctt gaa gaa atg ggt ttg att gat 4125 Leu Glu Gly Leu Cys Phe Leu Gln Leu Glu Glu Diet Gly Leu Ile Aap gtt ccg aag ctc act ctt gag tgt acc tca cag ttt cga gtc cgg tac 4173 Val Pro Lys Leu Thr Leu Glu Cys Thr Ser Gln Phe Arg Val Arg Tyr aaa ctc get gtc agc agt cct ata ata ctc sac aac atg ctc tca get 4221 Lys Leu Ala Val Ser Ser Pro Ile Ile Leu Asn Asn Met Leu Ser Ala gaa ggt ttc aca gtt cca gca cat ctc tcg ctt gaa gga tgt gag gaa 4269 Glu Gly Phe Thr Val Pro Ala His Leu Ser Lau Glu Gly Cys Glu Glu cca ttc att tca ttc gat gaa tcc gca aat ttc aca tct gtc aat aga 4317 Pro Phe Ile Ser Phe Asp Glu Ser Ala Asn Phe Thr Ser Val Asn Arg 1190 1195 1x00 ctg gaa ttc agt aat tgt gaa atg att tcc ctg cca sca aat ctg aag 4365 Leu Glu Phe Ser Asn Cys Glu Mat Ile Ser Leu Pro Thr Asn Leu Lys tgc ttc tcc act ctg cag aat ctt atg atc tgt gag tgc tac aac ata 4413 Cye Phe Ssr Thr Leu aln Asa Leu Diet Ila Cys Gllu C.'ys Tyr Asa Its 1x20 1225 1x30 tca tct tta cca gat ttg cca tcc tcc etc cag cac ata gaa ata att 4461 Ser Ser Leu 8ro Asp Leu Pro Ser Ser Leu Qla His Ile Cilu I1. Ile get tgt tct gat cgc ttg atg gag agc tgc cag gca cct gat ggt gaa 4509 Ala Cys Ser Asp Arg Leu Diet Qlu Ser Cys Qln Ala Pro Asp aly Glu agt tgg cca aag att gcg cat atc cgc tgg aag aca ttt gta 4551 Ser Trp Bro Lys Ila Ala His Ile Arg Trp Lys Thr Phe Val taaagtgtga ttcagatcct gacctacaaa taaacggaaa ggtaaattag ctcccatcag 4611 acaccttagc acccggaaag atatatagcc ttttctaata catgtgcatt tcgttaaaaa 4671 atttatcttt tcaatactct atttatttcc atccattgtg tacttcaaat aatggcagga 4731 cctccaggct ccagccacct atgacgagtt caagtgcctc agtctgtctc atcttgcagt 4791 tcaaccgggt taatgcagaa cctccaggac tgagaagttc aacatgcaat tattttgcgt 4851 gctccctttt cctggtgata tcagtgaaca atctggacat ggtgtcttgg ctgggcagca 4911 atgtgatgga tgatggcagc tgagaagtaa ttgcatgtgt taatgctata aggaacaatt 4971 agctacacag ttgacgcggt aatgctataa ggaacaatta gctacacagt tgacgcggta 5031 atgctatgga taaacaagcc actcatcgat gttgggagga gacatcgtcc tagtcaaaca 5091 agccactgag atcatgttgc ttagctttta ttcttttgaa gttttgtatg tccgaattct 5151 tctatgtcgc cgatgagtta gtcgaactcc aaggcgcttt gttacagtct gaaagatgat 5311 tagtttgcac ttctccgtcc agaattagtt gttgcagaaa tggatgtatc taaaggtaat 5371 tttagttcta gatacatcac aactaatttg ggacagaggg agtatcaagc tgtattgttt 5331 ctagcgactg ctgggatgag tggtccatta tctactggag aagttgcttt tcaaccaaaa 5391 aatgactgct gggatgagtg gtccattatc tactggagaa gttgcttttc aaccasaaaa 5451 taaacaaaaa atgtagtgga gcgatctatt aattacttaa tttgctggca aattgctctg 5511 tatattgatt gattgatcct atcaaataat actttttttg actcttcttt aattagagtt 5571 gtctgctata attggggtca gggaactgat aattcaattg ataaagtttt attttgcttt 5631 ggtatggacc gtctggaaag cccacaatga catgcacttt taaggttttt ttctgcatga 5691 ccaggcacga aatccgggac ctcctatatg acccagtttg cgtcatacag gagctcgacc 5751 cacagggtcg ac 5763 <210> 8 <211> 1279 <212> PRT
<Z13> Hordaum sp.
<400> 8 Met Ala Cilu Val Ala Leu Ala Ile Ala Ala Leu Arg Leu Ala Ala Leu Pro Val Leu Lys Lys Leu His Ala Asn Ala Ser Thr Tyr Leu Oily Val Asa Met Ala Arg Glu Ile His flu Leu f3lu Thr Thr Ile Met Pro Qln Phe alu Leu Val Ile (flu Ala Ala Asp Lys Oly Asn His Arg Pro Lys Leu Asp Lys Txp Leu (~ln (ilu Leu Lys alu 8er Phe Tyr Leu Ala 31u Asp Leu Leu Asp C~lu His alu Tyr Asn Ile Leu Lys His Lys Ala Lys Oly Lys Asp Ser Met Pro Ala Aen Oly Ser Sar Ile Ser Asn Thr Phe Met Lys Pro Leu Arg Ser Ala Sar Ser Arg Lau Ser Asa Leu Ser Ser 115 120 lay dlu Asn Arg Lye Leu Val Arg His Leu Lys Glu Leu Lys Ala Thr Leu Ala Lys Ala Lys Asp Phe His Qln Leu Leu Cys Leu Pro Ala f3ly Hie Asa Ala c3lu Arg Pro Ala Ile Pro Ser Asp Val Val Pro c3lu Ile Thr Ser Leu Pro Pro Mat Lye Val Ile Gly Arg Asp Lye Asp Arg Asp His Ile Ile Glu Cye Leu Thr Lye Val Thr Ala Thr Thr Glu Ser Ser Thr 195 Z00 a05 Thr Met Tyr Ser Gly Leu Ala Ile Val Gly Val Gly Gly Met Gly Lys a10 215 220 Ser Thr Leu Ala Gln Lsu Val Tyr Asn Asp Lys Arg Val Lys Glu Tyr Phe Asp Val Thr Mat Trp Val Ser Ile Ser Arg Lys Leu Asp Val Arg X45 a5o ass Arg His Thr Arg Glu Ile Ile Glu Ser Ala Ser Lys Gly Glu Cys Bro Arg Ile Asp Asn Leu Asp Thr Leu Gln Cys Lys Leu Thr Asp Ile Leu 275 a80 a85 Gln Glu Ser Gly Lys Phe Leu Leu Val Leu Asp Asp Val Trp Phe Glu Leu Gly Ser Glu Arg Glu Trp Asp Gla Lau Leu Ala Pro Leu Val Ser Arg Gln Thr Gly Ser Lys Val Leu Val Thr Ser Arg Arg Asp Thr Phe Pro Ala Thr Leu Cya Gys Glu Val Cys Pro Leu Glu Lys Met Asp Asp Ala Gln Phe Leu Ala Leu Phe Lys His His Ala Phe Ser Gly Pro Glu Ile Arg Asn Pro Gln Lau Arg Glu Lys Leu Glu Glu Phe Ser Lys Lys Ile Ala Lys Arg Leu Gly Gln Ser Pro Leu Ala Ala Lys Val Met Gly Ser Gln Leu Lys Gly Lya Thr Asp Ile Thr Ala Trp Lys Asp Ala Leu Thr Het Lys Ile Asp Lys Leu Ser Aap Pro Met Arg Ala Leu Leu Trp Ser Tyr Glu Lys Leu Asp Pro Cys Leu Gla Arg Cys phe Leu Tyr C,'ys Ser Leu Phe Pro Lys Gly His Lys Tyr Val Ile Asp Asp Leu Val His Leu Trp Met Ala Glu Gly Leu Val Asp Ser Cys Asn Gln Asn Lys Arg Val Glu Val Val Gly Arg Asp Cps Phe His Glu Met Ile Ser Val Ser Phe Phe Gln Pro Val Asp Glu Lys Tyr Thr Asp Thr Tyr Tyr Val Met His Asp Leu Leu His Asp Leu Ala Glu Ser Leu Ser Lys Glu Asp Tyr 515 5a0 525 Phe Arg Leu Glu Asp Asp Lys Val Thr Glu Ile Pro Ser Thr Val Arg His Leu Ser Val Arg Val Glu Sar Met Thr Gln His Lys Gln Ser Ile Cys Lys Leu His His Leu Arg Thr Ile Ile Cys Ile Asp Pro Leu Val Aep Asp Val Ser Asp Leu Phe Asn Gln Ile Leu Gln Asn Leu Lys Lys Leu Arg Val Leu Tyr Leu Ser Ser Tyr Ssr Sar Ser Gln Leu Pro Glu Ser Val Gly Glu Lau Lys His Leu Arg Tyr Leu Asn Ila Ile Gly Thr Leu Ile Ser Glu Leu Pro Arg Ser Leu Cys Thr Leu Tyr His Leu Gln Ser Leu Leu Leu Asn Asp Ser Val Lys Ser Leu Pro Glu Asn Ile Gars Asn Leu Arg Lys Leu Arg His Leu Glu Arg Asp Glu Leu Ala Leu Pro Gln Ile Pro Aan Ile Gly Lys Lau Thr Leu Leu Gln Gln Lau Asp Lys ~ 675 680 685 Phe Ser Val Gln Lys Lys Lye Gly Phe Glu Leu Glu Gln Leu Arg Asp Met Asa Glu Ile Arg Gly His Lsu Ser Val Glu His Leu Glu Asn Val Thr Gly Lys Asp Gln Ala Ile Glu Ssr Lys Leu Tyr Gln Lye Ser His 7a5 730 735 Leu Asp Ser Leu His Leu Gly Tsp Asn Leu Gly Asn Asn Thr Thr Ala Glu Asp Ser Leu His Leu Glu Its Leu Glu Gly Leu Thr Pro Pro Pro Gln Ile Ser Ala Leu 8er Ile Glu Gly Tyr Glu Ser Trp Lys Tyr Pro Gly Trp Leu Ile Asp Gly Ser Tyr Phe Glu Asn Leu Asn Tyr Leu Arg Phe Phe Gly Cys Arg Lys Leu Gln Ile Leu Pro Ser Asn Thr Glu Lsu Phe Val Asn Cys Thr Sar Leu Leu Lau Gln Gly Lsu Ser Asn Leu Asn Sa0 825 830 Thr Leu Pro Cys Leu Pro Leu Gly Leu Lys Val Leu Lye Val Gln Arg Cys Pro Leu Lau Ile Phe Ile Ser Hie Asp Glu Leu Glu His Asn Asg Gla Arg Glu Asn Ser Thr Arg Thr Asn His Lau Ala Ser Gla Leu Gly Leu Met Trp Glu Val Asp Ser Gly Ser Gly Ile Ser Thr Val Leu Leu Ser Glu Gars 8er Phe Leu Lys Gln Leu Met Ile Phe Thr His Ala Asp Met Ser His Val Gln Asn Phe Glu Ser Ala Lau Gln Arg Ala Lys Asn Gly Val Leu Val Lys Glu Asp Ila Ile Lys Ala Trp Ila Cys Cys His WO 99/45118 PC'T/AU99/00130 Glu Gln Ser Met Arg Leu Met Tyr Glu Arg Arg Ile Gly Leu Pro Leu Val Pro Pro Ser Gly Leu Hia Glu Leu His Leu 8er Ser Cys Ser Ile Thr Asp Glu Ala Leu Ala Val Cys Leu Asp Gly Leu Ala Ser Leu Gly Ser Leu Phe Lau Glu Lys Ile Phe Asn Leu Thr Lys Leu Pro Ser Alu Glu Val Leu Lys His Leu Ala Lys Leu Gly His Leu Asn Ile Thr Asp Cys Trp Cys Leu Arg Sar Leu Gly Gly Leu Arg Ala Ala Thr Ser Leu Ala His Phe Thr Leu Arg Ser Cys Pro Ser Leu Glu Leu Ala His Gly Ala Glu Cys Leu Pro Leu Ser Ile Glu Ser Leu Trp Ile Glu Lys Cys Met Leu Glu Gly Aan Phe Leu Cys Thr Asp Trp Pro His Met Asp Lys Its Ser Its Trp Aan Cys Arg Ser Thr Ala Cys Leu Ser Val Gly Ser Leu Thr Ser Val Lys Lys Lau Ser Leu Asp Arg Leu Bro Asp Lsu Gyre Met Leu Glu Gly Leu Cys Phe Leu Gln Leu Glu Glu Met Gly Leu Ile lla5 1130 1135 Asp Val Pro Lys Leu Thr Leu Glu Cys Thr Ser Gln Phe Arg Val Arg Tyr Lye Leu Ala Val Ser Ser Pro Ile Ile Lau Asn Asn Met Leu Ser Ala Glu Gly Phe Thr Val Pro Ala His Leu Ser Leu Glu Gly Cys Alu Glu Pro Phe Ile Ser Phe Asp Glu Ser Ala Asn Phe Thr Ser Val Asn Arg Leu Glu Phe Ser Asn Cys Glu Met Ile Ser Leu Pro Thr Asa Leu 1x05 1210 lal5 Lys Cys Phe Ser Thr Leu Gln Asa Leu Met Ile Cys Glu Gds Tyr Asn 1220 1a25 1230 Ile Ser Ser Leu Pro Asp Leu Pro Ser Ser Leu Gln His Ile Glu Ile 1235 1x40 1x45 Ile Ala Cys Ser Asp Arg Leu Met Glu Ser Cys Gln Ala Pro Asp Gly iZ50 1x55 1260 Glu Ser Trp Pro Lys Ile Ala His Ile Arg Trp Lys Thr Phs Val <a10> 9 <all> 7 <Z12> PRT
<a13> Artificial Sequence <ZZO>
<923> Description of Artificial 8equencespeptide <400> 9 Cys Phe Leu Tyr Cys Ser Leu <a10> 10 <211> 15 <al2> 8RT
<213> Artificial Sequence <aZ0>
<Za3> Description of Artificial Segueace:peptide <400> 10 Leu Gln Arg Cys Phe Lau Tyr Cys Ser Leu Phe Pro Lys Gly His <a10> 11 <211> 5 <zia> PRT
<213> Artificial Sequence _ <ZZO>
<Z23> Description of Artificial Sequence: peptide <400> 11 Trp Xaa Ala Glu Gly <Z10> 12 <all> 10 <ala> PRT
<213> Artificial Sequence <aZ0>
<aZ3> Description of Artificial Sequence: peptide <400> 12 Glu Leu Val His Lau Trp Xaa Ala Glu Gly <910> 13 <211> 9 <ala> pRT
<a13> Artificial Sequence <ZZO>
<223> Description of Artificial Seguencespeptide <400> 13 Val Gly Xsa Gly Gly Xaa Gly Lye Ser <210> 14 <all> 19 <212> BRT
<a13> Artificial Sequence <aao>
<2a3> Description of Artificial Sequence: peptide <400> 14 Tyr Ser Gly Lau Ala Ile Val Gly Xaa Gly Gly Xaa Gly Lys Ser Xaa Leu Ala Gln <alo> 15 <all> 7 <a12> PRT
<a13> Artificial Sequence <aao>
<aa3> Description of Artificial Sequsnce:peptide <400> 15 Leu Leu Val Leu Asp Asp Val <a10> 16 <all> la <ala> PRT
<a13> Artificial Sequence <aa0>
<aa3> Description of Artificial 8equance:peptide <400> 16 Lye Phe Leu Leu Val Leu Asp Asp Val Trp Phe alu <a10> 17 <all> 9 <a12> PRT
<a13> Artificial Sequence <aao>
<aa3> Description of Artificial Seguence:peptide <400> 17 Lys Tyr Ser Gly Leu Ala Ile Val Oly <a10> 18 <all> 9 <ala> PRT
<a13> Artificial Sequence <aao>
<aa3> Description of Artificial Sequence: peptide <400> 18 Tyr Ser Gly Leu Ala Ile Val Gly Leu <Z10> 19 <Z11> 7 <212> pRT
<Z13> Artificial 8equeace <aao>
<a23> Description of Artificial Seguence:peptide <400> 19 Leu Gly Gly Met Gly Lys 8er <210> 20 <all> 6 <Z12> BRT
<a13> Artificial Sequence <ZZ0>
<223> Description of Artificial Segusnce:peptide <400> 20 Gly Gly Met Gly Lys Sar <210> Zl <Z11> 7 <a12> PRT
<213> Artificial 8equeace <ZZO>
<223> Description of Artificial 8equence:peptide <400> 21 Gly Gly Met Gly Lye 8er Thr <alo> az <all> to <ala> PRT

WO 99/4511$ PCT/AU99/00130 <213> Artificial Sequence <aao>
<223> Description of Artificial Sequsace:peptide <400> 22 Gly Gly Dset Gly Lye Ser Thr Leu Ala Gla <210> 23 <211> 7 <212> PRT
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:peptide <400> 23 Gly Lys Ser Thr Leu Ala Gla <210> 24 <211> 5 <212> PRT
<213> Artificial Sequence <220>
<223> Description of Artificial Sequeacespeptide <400> 24 Ser Thr Leu Ala Gla <210> 25 <211> 5 <212> PRT
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence: peptide <400> 25 Thr Leu Ala Gla Tyr <a10> a6 <all> 13 <ala> PRT
<a13> Artificial Sequence <aao>
<aa3> Description of Artificial Seguance:peptide <400> a6 aln Lys Phe Leu Leu Val Lau Asp Asp Val Trp Pha alu <al0> a7 <all> 13 <ala> PRT
<a13> Artificial Sequaaca <aao>
<aa3> Description of Artificial 8aquanca:paptide <400> a7 Lys Phe Leu Lau Val Leu Asp Asp Val Trp Phe (flu Lys <a10> a8 <all> is <ala> PRT
<a13> Artificial Sequence <aao>
<aa3> Description of Artificial Saguence:paptide <400> a8 Arg Leu aln Arg Cys Pha Leu Tyr Cys 8er Lau Phe Pro Lys c~ly His <a10> a9 <all> 16 <a12> PRT
<a13> Artificial Sequence <aa0>
<2a3> Description of Artificial Segueace:peptide <~oo> as Lau Gln Arg Cya Phe Leu Tyr Gars Sar Leu Phe Pro Lys Gly His Arg <a10> 30 <a11> 10 <a12> PRT
<a13> Artificial Sequence <aa0>
<aa3> Description of Artificial Saquence:peptide <400> 30 Glu Leu Val His Leu Trp Val Ala Glu Gly <a10> 31 <al1> 5 <ala> PRT
<a13> Artificial Saqueace <aa0>
<aa3> Dascriptioa of Artificial Sequsnce:peptida <400> 31 Trp Val Ala Glu Gly <a10> 3a <211> 11 <ala> PRT
<a13> Artificial Sequence <aa0>
<aa3> Description of Artificial Sequance:peptida <400> 3a Aen Glu Leu Val Hie Leu Trp Val Ala Glu Gly <a10> 33 <a11> 11 <a1a> PRT
<a13> Artificial Sequence 6a <a~0>
<Z23> Description of Artificial 8equeace:peptide <400> 33 Qlu Leu Val His Leu Trp Val Ala cilu aly Phe <Z10> 34 <Z11> 25 <a12> DNA
<Z13> Artificial 8eguence <Za0>
<Z23> Description of Artifiaisl Sequence:oligoaucleotide primer <400> 34 aagaattcgg agtaggaaaa acaac 25 <310> 35 <Z11> 25 <ala> DNA
<Z13> Artificial Sequence c2Z0>
<Za3> Description of Artificial 8equeace:oligoaucleotide primer <400> 35 aagaattcgg agtaggnaaa actac 25 <Z10> 36 <211> 25 <a12> DNA
<a13> Artificial Sequence <Za0>
<a23> Description of Artificial 8eguence:oligonucleotide primer <400> 36 aagaattcgg agtnggaaaa accac 25 <210> 37 <all> 25 <212> DNA
<213> Artificial Sequence <ZZO>
<2a3> Description of Artificial Segueacasoligonucleotide primer <400> 37 sagaattcgg ngtnggnaaa acgac 25 <210> 38 <all> Z5 <ala> DNA
<Z13> Artificial Sequence <aao>
<a23> Description of Artificial Sequence:oligonucleotide primer <400> 38 aagaattcgg ngtnggnaag acaac a5 <210> 39 <Z11> 25 <212> DNA
<a13> Artificial Sequence <aao>
<223> Description of Artificial Sequence:oligonucleotide primer <400> 39 aagaattcgg agtnggnaag actac 25 <a10> 40 <all> 25 <ala> DNA
<a13> Artificial Sequence <aao>
<2a3> Description of Artificial Seguence:oligonucleotide primer <400> 40 aagaattcgg ngtaggaaag accac 25 <210> 41 <211> 25 <212> DNA
<213> Artificial Sequeace <220>
<223> Description of Artificial Sequeace:oligonucleotide primer <400> 41 aagaattcgg ngtnggasag acgac 25 <210> 42 <211> 20 <212> DNA
<213> Artificial Sequence <220>
<Z23> Description of Artificial Sequence:oligonucleotide primer <400> 42 ctactgatac tagacgacgt 20 <210> 43 <211> 20 <212> DNA
<213> Artificial Sequence <220>
<223> Description of Artificial Sequeace:oligoaucleotide primer <400> 43 ctactgatac tagacgatgt 20 <21D> 44 <211> 20 <212> DNA
<213> Artificial Segueace <aa0>
<aa3> Description of Artificial Sequeace:oligonucleotide primer <400> 44 ctactgntnc tngatgacgt a0 <a10> 45 <ail> a0 <ala> DNA
<a13> Artificial Sequence <aao>
<aa3> Description of Artificial Sequence:oligoaucleotide primer <400> 45 ctactgntnc tngatgatgt a0 <a10> 46 <all> 25 <ala> DNA
<a13> Artificial Sequence <aa0>
<aa3> Description of Artificial Sequsnce:oligonucleotide primer <400> 46 aactcgagag ngcnagnggn aggcc 25 <a10> 47 <all> a5 <ala> aNA
<a13> Artificial Sequence <aao>
<aa3> Deacriptioa of Artificial Sequence:oligonuclsotide primer <400> 47 aactcgagag ngcnagnggn agacc a5 <a10> 48 _ <111> Z5 <Z12> DNA
<213> Artificial 8equeace <2Z0>
<223> Dascriptioa of Artificial Seguencssoligonucleotide primer <400> 48 aactcgagag ngcnagnggn agtcc 25 <Z10> 49 <211> 25 <Zla> DNA
<213> Artificial 8eguenca <220>
<223> Description of Artificial 8eguence:oligonucleotide primer <400> 49 aactcgagag agcnagnggn agccc ~ 25 <~10> 50 <Z11> Z5 <212> DNA
<Z13> Artificial Saqueace <a20>
<Z13> Description of Artificial Sequencasaligonucleotide primer <400> 50 aactcgagaa ngccaanggc aatcc a5 <Z10> 51 <Z11> 25 <21a> DNA
<Z13> Artificial Segueace <Za0>
<223> Des3cription of Artificial 8equencasoiigonucleotide primer <400> 51 aactcgagaa agccaaaggc aaacc 25 <210> 52 <all> ZO
<ala> DNA
<213> Artificial Sequeace <aZ0>
<Z23> Descriptioa of Artificial Segoeace:oligoaucleotide primer <400> 52 carviangcra arcaytgttt 20 <210> 53 <~11> ao <ala> DNA
<213> Artificial Sequence <Z20>
<a23> Description of Artificial Sequeace:oligoaucleotide primer <400> 53 carwaagcra arcaytgctt 20 <210> 54 <211> a0 <21Z> DNA
<Z13> Artificial Segusace <aao>
<223> Description of Artificial Sequencs:oligonucleotide primer <400> 54 atagarcarw aagcraaaca 20 <a10>55 <all>ao <Z12>DNA

<213>Artificial Segueace <aao>
s8 <aa3> Description of Artificial 8eguencesoligonucleotide primer <400> 55 atagarcarw angcraagca 20 <210> 56 <Z11> Z0 <a1a> DNA
<z13> Artificial Sequence <aao>
<a23> Description of Artificial Sequence:oligonucleotide primer <400> 56 ayraancent nagccatcca ZO
<210> 57 <a11> ZO
<a1a> DNA
<213> Artificial Sequence <ZZO>
<223> Description of Artificial Sequence:oligonucleotide primer <400> 57 ayraanccnt ntgccatcca 20 <210> 58 <211> 20 <212> DNA
<Z13> Artificial Sequence <aao>
<Z23> Description of Artificial Sequence:oligonucleotide primer <400> 58 ayraanccnt ncgccatcca 20 <210> 59 <a11> ao <212> DNA
<213> Artificial Sequence <ZZO>
<223> Description of Artificial Sequence:oligonucleotide primer <400> 59 ayraanccnt nggccatcca 20 <210> 60 <211> 18 <Z12> DNA
<Z13> Artificial Sequence <220>
<Z23> Description of Artificial Sequencesoligonucleotide primer <400> 60 cgacagtcna tcatgcst 18 <210> 61 <211> 18 <a1a> DNA
<213> Artificial Sequence <aao>
<223> Description of Artificial Sequence:oligonucleotide primer <400> 61 cgacagtcna tcgtgcat 18 <Z10> 62 <211> 18 <a12> DNA
<213> Artificial Sequence <aao>
<Z23> Description of Artificial Sequence:oligonucleotide primer <400> 62 cgacagtcng tcatgcat 18 <a10> 63 <all> 18 <ala> DNA
<a13> Artificial 8egueace <aa0>
<aa3> Description of Artificial Sequence:olfgonucleotide primer <400> 63 cgacagtcng tcgtgcat 18 <a10>64 <all>3855 <ala>DNA

<a13>Zea ways <400> 64 atggccgact tcgcgctcgc cggcttaagg tgggcagcat cgccgattgt caacgagctt 60 cttactaaag cttcagctta cctcagtgtg ggcatggtgc gtgagatcca acgactagaa la0 gccactgtcc tgccacagtt cgagctggtg attcaagcgg cccagaagag cccccacagg 180 ggcatactgg aggcatggct ccggcgtctc aaagaagcct actatgatgc cgaggacttg a40 ttggacgagc atgagtacaa tgtccttgag ggcaaggcca agagcggasa aagtctcctg 300 ctgggagagc atggaagctc ctccactgca actactgtca tgaagccttt tcatgctgct 360 atgagcaggg cacgcaactt gctccctggg aacagaaggc taattagcaa gatgaacgag 4a0 ctcaaagcta ttctgacaga agcccaacaa cttcgagatc ttcttggctt gccacatggc 480 aataccgtcg agtgcccagc tgcagcacct atcagtgttc ccacaaccac atcacttccc 540 acttccaagg tttttggtcg cgacagggat cgtgatcgta tagtagattt tcttctcggc 600 aagscaacaa ctgctgaggc aagctcagct aagtacctcg gtttggccat tgttggattg 660 ggaggaatgg ggaagtccac cttagcacag tatgtctata atgacaaaag gatagaagaa 7a0 tgctttgata tcaggatgtg ggtgtgcatc tcacgcaaac ttgatgtgca tcgtcacaca 780 gggagattat ggagtctgca aaaaagggag agtgcccacg tgttgataat ctcgatactc 840 tgcagtgcaa attacgtgat atactacaag agtcacagaa attcctgctt gtcttggstg 900 atgtttggtt tgaaaaatct cataatgaga cagagtggga gttattcctt gctcccttag 960 tctctaaaca gtcagggagc aaagttttgg tgacttctcg aagtaaaact cttcctgccg 1020 ctatttgttg tgaacaaaaa catgtcattc atttgaaaaa catggatgat actgagtttt 1080 tggctctttt taaacsccat gctttctctg gagcagaaat caaagaccaa ctattacgca 1140 cgaagctgga agacactgca gtggsgattg ctaaaaggct tggacaatgt cctttggcag 1x00 caaaagttct gggttctcga ttgtgcagga aaaaggatat tgctgaatgg aaagctgctc 1x60 taaagattgg agatttaagt gatcccttca catctctgtt gtggagttac gagaagttag 13x0 atccacgtct gcagaggtgc ttcttgtatt gcagcttgtt tccaaaaggt catagatatg 1380 aacctaatca gttggttcac ctctgggtgg cagaaggatt tgttggttca tgcaatttga 1440 gtaggagaac attggaagag gctgggatgg attacttcaa tgatatggtc tctggatcat 1500 tcttccaasg gtacggtggt ggtcggtact atgtcatgca tgatatcctt catgattttg 1560 cagagtcact ctctagagaa gactgcttta gattagaaga tgataatgtg acagaaatac 1620 catgcactgt tcgacatcta tctgttcatg ttcaaagtat gcaaaagcat aagcaaatta 1680 tctgcaagct atatcattta cgcactatta tctgcatcga tccgctaatg gstggcccaa 1940 gtgatatttt tgatggcatg ctacggaacc aaagaaaatt gcgtgtattg tctctgtcat 1800 tttscagcag caacaagttg ccagaatcta ttggtgagct gaagcacctc cggtatttga 1860 acctcatcag gacgttagtt tctgaattgc ctacatcatt atgtactctc taccacttac 1920 aattactttg gttaaaccac atggtggaga atttgcctga caaactatgc aatttaagaa 1980 agctacgaca tctaggagcg tacgtgaatg atttcgcgat tgaaaagcct atttgccaaa x040 ttctgaatat aggtaagtta acgtcgctac aacacattta tgtcttttct gtacaaaaga 2100 agcaaggcta tgagttgcga cagttgaagg acttgaatga gcttggtggc agtttaaaag 2160 tgaaaaatct tgagaatgtc attggaaagg atgaagccgt agagtcgaag ctatatctga 2x20 aaagtcgcct taaagagtta gcatttgagt ggsgttccga gsatggcatg gatgcaatgg aa80 ~a atattctaga aggtctgaga ccgccscccc aactgagtaa gctcacaatc gaaggttaca 2340 gatctgatac atatcctggg tggttactag agcgatccta ttttgagaat ttggaaagtt 2400 ttgagcttag taattgcagt ttgctagaag gcctaccacc agatacagag ctccttcgga 2460 attgctctag gttgcgtata aacattgttc caaatttgaa ggaactatct aatcttccag 2520 caagccttac agatttatca attgattgtt gcccactgct tatgtttatc accaacaatg 2580 agctaggaca gcatgacttg agggaaaata taataatgaa ggcagacgac ctggcatcta 2640 aacttgcatt gatgtgggag gtggattcag gaaaagaagt taggaatata ctgtcgaaag 2700 actattcatc tctgaagcag ttgatgacat tgatgatgga tgatgatata tcaaagcatc 2760 ttcaaattat tggaagtggt ctggaggaaa gagaagataa agtatggatg aaagaaaaca 2820 tcatcaaggc atggctcttt tgccatgaac agaggataag attcatttat ggaaggacca 2880 tggagatgcc attggttcta ccatcaggac tatgtgaact ttctctttct tcatgcagta 2940 ttacagatga agctttagct atttgccttg gtggcctcac ttcactgggg tatttaaact 3000 tgagatataa tatggaatta actacacttc catcagaaaa ggtgtttgag catttgacaa 3060 agcttgacac gttgattcta aatggttgtt ggtgtctcaa gtcactgggt ggcttacgtg 3120 atgctccatc tctttcctat tttaactatt gggattgtcc ttctttagag ctagcacgtg 3180 gagcagaact aatgccgttg aaccttgcta tcagtctcag catccgtggc tgcattcttg 3240 cagctgattc gttcattaat ggcttgccac acctgaaacg tctttacatt aaagtctgca 3300 gaagctcccc atccttatcg attggccacc tgacctccat tgaatcatta cgtctaaatg 3360 gtctccctga tctttacttt gttgaaggct tgtcttccct gcaccttaag cacctacatt 3420 tagtagatgt tgcaaacctc actgccaagt gcatctcaca gtttcgtgtc caggaatcgc 3480 tcacggttag tagctctgta ttgctcaacc acatgctaat ggctgaaggg tttacagccc 3540 caccaaatct tactctttta gattgcaagg agccgtcagt ttcatttgaa gaacctgcaa 3600 atctctcatc cgtcaagcac ctgcactttt catgttgcga aacagagtcc ctgcctagaa 3660 atctaaaatc tgtctcaagt ctggagagtc tttctataga acattgcccc aacataacat 3720 ctttaccaga tctgccgtcc tccctccagc gcataactat atgggattgt cccgtcttga 3780 tgsagaattg ccaagaacct gatggagaaa gctggccaaa gatttcacac gttcgctgga 3840 agagctttct actaa 3855 <a10> 65 <all> 3879 <Z12> DNA
<213> Zea maye <400> 65 atggccgact tcgcgctcgc cggcttaagg tgggcagcat cgccgattgt caacgagctt 60 cttactasag cttcagctta cctcagtctg gacatggtgc gtgagatcca acgactagaa 1a0 gccactgtcc tgccacagtt cgsgctggtg sttcaagcgg cccagaagag cccccacagg 180 ggcatactgg aggcatggct ccggcgtctc aaagaagcct actatgatgc cgaggacttg a40 ttggacgagc atgagtacaa tgtccttgsa ggcaaggcca agagcggaaa aagtctcctg 300 ctgggagagc atggaagctc ctccactgcs actactgtca cgaaaccttt tcatgctgcc 360 atgagcaggg cgcggaactt gctccctcaa ascagaaggc taattagcaa gatgaacgag 4a0 ctcaaagcaa tcctgacaga agcccaacaa cttcgagatc ttcttggttt gccacatggc 480 aataccatcg ggtggccagc tgcagcacct accagtgttc ccacaaccac atcacttccc 540 acttccaagg tttttggtcg cgacagggat cgtgatcgta tsgtsgattt tcttctcggc 600 aagacaacaa ctgctgaggc aagctcagct aagtactcgg gtttggccat tgttggattg 660 ggaggaatgg ggaagtccac cttagcacag tstgtctata atgacaaaag gatagaagaa 720 tgctttgata tcaggatgtg ggtgtgcatc tcacgcaaac ttgatgtgca tcgtcacacs 780 agggagatta tggagtctgc aaaaaaggga gagtgcccac gtgttgataa tctcgatact 840 ctccagtgca aattacgtga tstactacaa gagtcacaga aattcctgct tgtcttggat 900 gatgtttggt ttgaaaaatc tcataatgag acagagtggg agttattcct tgctccctta 960 gtctctaaac agtcagggag caaagttttg gtgacttctc gaagtaaaac acttcctgcc 1020 gctatttgtt gtgaacaaga acatgtcatt catttggaaa acatggatga tactgagttt 1080 ttggctcttt ttaaacacca tgctttctct ggagcagaaa tcaaagacca actgttacgc 1140 acgaagctgg aagacactgc agaggagatt gctaasaggc ttggacaatg tcctttggca 1200 gcaaaagttc tgggttctcg attgtgcagg aaasaggata ttgctgaatg gaaagctgct 1260 ctaaagcttg gagatttaag tgatcccttc acatctctgt tgtggagtta cgagaagtta 1320 gatccacgtc tgcagaggtg cttcttgtat tgcagcttgt ttccaaaagg tcacggatat 1380 agacctgaag agttggttca cctttgggtg gcagaaggat ttgttggttc atgcaatttg 1440 agtaggagaa cattggaaga ggctgggatg gattacttca atgatatggt ctctggatca 1500 ttcttccaaa ggtacggtcg gtactatgtc atgcatgata tccttcatga ttttgcagag 1560 tcactctcta gagaagactg ctttagatta gaagatgata atgtgacaga aataccatgc 1620 actgttcgsc atctatctgt tcstgttcaa agtatgcaaa agcataagca aattatctgc 1680 asgctatatc atttacgcac tattatctgc stcgatccgc taatggatgg cccaagtgat 1740 atttttgatg gcatgctacg gaaccaaaga aaactgcgtg tattgtctct gtcattttsc 1800 aacagcagca agttgccaga atctattggt gagctgaagc acctccggta tttgaacctc 1860 atcaggacat tagtttctga attgcctaca tcsttatgta ctctctacca cttacsatts 19x0 ctttggttaa accacatggt ggagaatttg cctgacasac tatgcaattt aagaaagcta 1980 cgacatctag gagcgtactc atcgtacgct aatgattccg tgaatgaaac gcctatttgc 2040 caaattctga atataggtaa gttaacgtcg ctacaacaca tttatgtctt ttatgtacag 2100 aagaagcaag gttstgagtt gcgacagatg aaggacttga atgagcttgg tggcagttta 2160 atagtgaaaa atcttgagaa tgtcattaga aaggatgaag ccgtagagtc gaagctatat ZZZO
ctgsaaagtc gccttaaaga gttggcactt gagtggagtt ccgagaatgg catggatgca 2280 atggatattc tagaaggtct gagaccgcca ccccaactga gtaagctcac aatcaaaggt 2340 tacagatctg atacatatcc tgggtggtta ctagagcgat cctattttga gaatttggaa 2400 agttttgagc ttagtaattg csgtttgcta gaagtcctac caccagatac agagctcctt 2460 cggaattgct ctaggttgca tataaacttt gttccaaatt tgaaggaact atctaatctt 2520 ccagcaggcc ttacagattt atcaattgat tgttgcccac agcttatgtt tatcaccaac 2580 aatgagctag gacagcatga cttgagggaa aatstaataa tgaaggcaga cgacctggca x640 tctaaacttg cattgatgtg ggaggtggat tcaggaaaag aagttatgag agtactgtcg 2700 aaagactatt tatctctgaa gcagttgatg acattgstga tggatgatga tatatcaaag 2760 catcttcaaa ttattggaag tggtctgaag gaaagagaag ataaagtttg gatgaaagaa Z8Z0 aacatcatca aggcatggct cttttgccat gagcagagga taagattcat ttatggaagg 2880 accatggaga tgccattggt tctaccgtca ggactctgtg aactttctct ttcttcatgc 2940 agtattacag atgaagcttt agctatttgc cttggtggcc tcacttcact gagaaattta 3000 cgattggaat ataatatggc attaactaca cttccatcag aaaaggtgtt tgagcatttg 3060 acaaagcttt acaggttggt tgtaagsggt tgtttgtgtc tcaaatcact ggggggctta 3120 cgtgctgctc catctctttc ctgttttgac tgttcggstt gtcctttttt agagctagca 3180 cgtggagcag aactaatgcc gttgaacctt gctggagacc tcaacatccg tggctgcatt 3240 cttgcagttg attcattcat taatggcttg ccacacctga aacatctttc catttatttc 3300 tgcagaagct ccccatcctt atcgattggc cacctgacct cccttcaatc attagatcta 3360 atggtctgcc tgatctttac tttgttgaag gcttgtcttc cctgcacctt aagcacctac 3420 gtttagtaga tgttgcaaac ctcactgcca agtgcatctc accgtttcgt gtccaggaat 3480 ggctcacagt tagtagctct gtattgctca accacatgct aatggctgaa.gggtttacag 3540 tcccaccaaa acttgttctt ttctgttgca aggagccgtc agtttcattt gaagaacctg 3600 caaatctctc atccgtcaag cacctgcact tttcatgttg tgaaacaaag tccctgccga 3660 gaaatctaaa atctgtctca agtctggaga gtctttctat aaacggttgc cccaacataa 3720 catctttacc agatctgccg tcctccctcc agcgcataac tttattggst tgccccgtct 3780 tgatgaagaa ctgccaagaa cctgatggag aaagctggcc aaagsttcta cacgttcgct 3840 ggaagagctt tctaecaata tcgatctttt tttttttag 3879 <ZlOa 66 <211> 3837 <Z12> DNA
<a13> Zaa a~aye <400a 66 atggccgact tggcgctcgc cggcttsagg tgggcagcat cgccgattgt caacgagctt 60 cttactaasg cttcagctta cctcagtgtg gacatggtgc gtgagatcca acgactagaa 120 gccactgtcc tgccacagtt cgagctggtg attcaagcgg cccagaagag cccecacagg 180 ggcatactgg aggcatggct ccggcgtctc aaagaagcct actatgatgc cgaggacttg 240 ttggacgagc atgagtacaa tgtccttgaa ggcasggcca agagcggaaa aagtctcctg 300 ctgggagagc atggaagctc ctacactgca actactgtca cgaaaccttt tcstgctgcc 360 atgagcaggg cgcggaactt gctccctcaa aacagaaggc taattagcaa gatgaatgag 420 ctcsaagcaa tcctgacaga agcccaacaa cttcgagatc ttcttggttt gccacatggc 480 aataccatcg ggtggccagc tgcagcacct accagtgttc ccacaaccac atcacttccc 540 acttccaagg tttttggtcg cgacagggat cgtgatcgta tagtagattt tcttctcggc 600 aagacsacaa ctgctgaggc aagctcagct aagtactcgg gtttggccat tgttggattg 660 ggaggaatgg ggaagtccac cttagcacag tatgtctata atgacaaaag gatagaagaa 720 tgctttgata tcaggatgtg ggtgtgcatc tcacgcaaac ttgatgtgca tcgtcacaca 780 agggagatta tggagtctgc aaaaaaggga gagtgccgac gtgttgataa tctcgstact 840 ctccagtgca aattacgtga tatactacaa gagtcacaga aattcctgct tgtcttggat 900 gatgtttggt ttgaaaaatc tcataatgag acagagtggg agttattcct tgctccatta 960 gtctctaaac agtcagggag caaagttttg gtgacttctc gaagtaaaac acttcctgcc lOZO
tctatttgtt gtgaacaaga acatgtcatt catttggaaa acatggatga tactgagttt 1080 ttggctcttt ttaaacacca tgctttctct ggagcagaaa tcaaagacca actgttacgc 1140 acgaagctgg aagacactgc agaggagatt gctaaaaggc ttggacaatg tcctttggca 1x00 gcaaaagttc tgggttctcg attgtgcagg aaaaaggsta ttgctgastg gaaaactgct 1260 WO 99/45118 PC'TIAU99/00130 ctaaagattg gagatttaag tgatcccttc acatctctgt tgtggagtta cgagsagtta 1320 gatccacgtc tgcagaggtg cttcttgtat tgcagcttgt ttccaaaagg tcacgtatat 1380 agacctcaag agttggttca cctttgggtg gcagaaggat ttgttggttc atgcastttg 1440 agtaggagaa cattggaaga ggctgggatg gattacttca atgatatggt ctctggatca 1500 ttcttccaat ggtacggtcg gtactatgtc atgcatgata tccttcatga ttttgcagag 1560 tcactctcta gagaagactg ctttagatta aaagatgata atgtgacaga aataccatgc 16x0 actgttcgac atctatctgt tcatgttcaa agtatgcaaa agcatasgca aattatctgc 1680 aagctatatc atttacgcac tattatctgc ctcgatccgc taatggatgg cccaagtgat 1740 atttttgatg gcatgctacg gaaccaaaga aaactgcgtg tattgtctct gtcattttac 1800 aacagcagca agttgccaga atctattggt gagctgaagc acctccggta tttgaacctc 1860 atcaggacgt tagtttctga attgcctaca tcattatgta ctctctacca cttacaatta 1920 ctttggttaa accacatggt ggagaatttg cctgacaaac tatgcaattt aagaaagcta 1980 cgacatctag gagcgtacaa atggtacgct catggtttcg tggaagaaat gcctatttgc 2040 caaattgtgs atataggtaa gttaacgtcg ctacaacaca tttatgtctt ttctgtacag 2100 aagsagcaag gttatgagtt gcgacagttg aaggacttga atgagcttgg tggcagttta 2160 agagtgaaas atcttgagaa tgtcattgaa aaggatgaag ccgtagagtc gaagctatat ZZZO
ctgaaaagtc gccttaaaga gttggcactt gagtggagtt ccaagaatgg catggatgca x280 atggatattc tagaaggtct gagaccgcca ccccaactga gtaagctcac aatccaaggt x340 tacggatctg atacatatcc tgggtggtta ctagagcgat cctattttga gaatttggaa x400 agttttgagc ttattaattg cagattgcta gaaggcctac caccagatac agagctcctt x460 cggaattgct ctaggttgca tatsasctct gttccaaatt tgaaggaact atctaatctt 2520 ccagcaggcc ttacagattt atcaattgat tgttgcccac tgcttatgtt tatcaccaac 2580 aatgagctag gacagcatga cttgagggaa aatataataa tgaaggcaga cgccctggca 2640 tctaaacttg cattgatgtg ggaggtggat tcaggattta gtgttagcag tgtgctgtgg ZT00 WO 99/45118 PC'T/AU99/00130 gaaagactat tcatctctta agcatcttca aattattgaa actggtctag aggaaggaga x760 taaagtatgg atggaagaaa acatcatcaa gccatggctc ttttgccatg agcaaggata 2820 agattcattt atggaaggac catggagatg ccattggttc taccgtcagg actctgtgaa 3880 ctttctcttt cttcatgcag tattacagat gaagctttag ctatttgcct tggtggcctc 2940 acttcactga gaactttaca attggaatat aatatggcat taactacact tccatcagaa 3000 aaggtgtttg agcatttgac aaagcttgsc aggttggttg taagaggttg tttgtgtctc 3060 aaatcactgg ggggcttacg tgctgctcca tctctttcct gttttgactg ttcggattgt 31x0 ccttttttag agctagcacg tggagcagaa ctaatgccgt tgaaccttgc tggagacctc 3180 aacatccgtg gctgcattct tgcagttgat tcattcatta atggcttgcc acscctgaaa 3x40 catctttcca tttatttctg csgaagctcc ccatccttat cgattggcca cctgacctcc 3300 cttcaatcat tagatctata tggtctgcct gatctttact ttgttgaagg cttgtcttcc 3360 ctgcacctta agcacctacg tttagtagat gttgcaaacc tcactgccsa gtgcstctca 3420 ccgtttcgtg tccaggaatg gctcacagtt agtagctctg tattgctcaa ccacatgcta 3480 atggctgaag ggtttacagc cccaccaaat cttactcttt ttgtttgcaa ggagccgtca 3540 gtttcatttg aagaacctgc aaatctctca tccgtcaagc acctgctgtt ttcatgttgc 3600 aaaacagsgt ccctgccgag asstctaaaa tctgtctcaa gtctggagag tctttctata 3660 cacagttgcc ccaacataac atctttacca gatcttccgt cctccctcca gctcatacgt 3720 atatcagatt gccccgtctt gaagaagaac tgccaagaac ctgatgggga aagctggcca 3780 aagatttcga accttcgctg gaagcacatt ctactaatac caaacttgct tctttag 3837 <210> 67 <all> 3833 <21Z> DNA
<213> Zea mat's <400> 67 atggcggact tggcgctagt tggcttaagg tgggcagcat cgccgattgt caaggagctt 60 cttactaaag cttcagctta cctcagtgtg gacatggtgc gtgagatcga acgactacaa 1a0 gacactgtcc tgccacagtt cgagttggtg attcaagcgg cccagaagag cecccatagg 180 ggcaagctgg satcctggct tcggcgtctc aaagaagcct tctatgatgc cgaggacctg 240 ctggacgagc atgagtacaa cgtccttaag gccaaggcca agagcggaaa aggtcccctg 300 etccgagagg atgaaagctc ctccactgca accactgtca tgaagccttt tcattctgct 360 atgaacaggg cacgcaactt gctccctggg aacagaaggc taattagcaa gatgaacgag 4Z0 ctcaaagcta ttctgscaga agccaagcag cttcgagatc ttcttggctt gccacatggc 480 aataccgtcg agtggccagc tgcagcacct accagtgttc ccacaaccac atcacttccc 540 acttccaagg ttttcggtcg cgacagggat cgtgatcgca tagtaaaatt tcttctcggc 600 aagacaacaa ctgcagaggc sagctcaact aagtactccg gtttggccat tgttggattg 660 ggaggaatgg ggasgtctac cttagcacaa tatgtctata atgacaagag gattgaagaa 7Z0 tgctttgatg tcaggatgtg gatctgtatc tcgcgcaaac ttgatgtgca tcgtcacaca 780 agggagatca ttgagtccgc aaaasagggg gagtgcccac gtgtcgataa tctcgatact 840 ctccagtgca aactacgaga catactacaa cagtcaaasa sattcctgct tgtcttggat 900 gatgtttggt ttgaaaaatc tgatagtgag acagagtggg acctactcct tgctccatta 960 gtctctaaac agacgggaag cagagttttg gtgacttctc gacgtgaaat gcttccagcc lOZO
gctgtttgct gtgaacgagt tgttcgtttg gaaaacatgg atgatactga gttcttggct 1080 ctctttaaac aacatgcttt ctctggsgca aaaatcaaag accagctgtt acgcacgaag 1140 ctggaacata ctgcagggga gcttgctaaa aggcttggac satgtccttt ggcagcaaaa 1200 gttctgggtt cccgattgtg caggaaaaag gatattgctg aatggaaagc tgctctaaag iZ60 cttggagatt taagtgatcc ettcacatct ctgttgtgga gttacgagaa gttagatcca 1320 cgtctgcaga ggtgcttctt gtattgcagc ttgtttccaa aaggtcatag atatgsacct 1380 aatgagttgg ttcacctctg ggtggcagaa ggatttgttg gttcatgcaa tttgagtagg 1440 agaacattgg aagaggctgg gatggattac ttcaatgata tggtctctgg atctttcttc 1500 caaaggtacc gtcggtacta tgtcatgcat gatatccttc atgattttgc agagtcactc 1560 tctagagaag actgctttag attagaagat gataatgtga cagaaatacc atgcactgtt 16x0 cgacatctat ctgttcatgt tcaaagtatg caaaagcata agcaaattat ctgcaagcta 1680 tatcatttac gcactattat ctgcatcgat ccgctaatgg atggcccaag tgatattttt 1740 gatggcatgc tacggaaccg aagaaaactg cgtgtattgt ctctgtcatt ttacaacagc 1800 agcaagttgc cagaatctat tggtgagctg aagcacctcc ggtatttgaa cctcatcagg 1860 acgttagttt ctgaattgcc tacatcstta tgtactctct accacttaca attactttgg 19x0 ttaaaccaca tggtggagaa tttgcctgac aaactttgca stttaagaaa gctacgacat 1980 ctaggagcgt acacatggaa agaaaagcct atttgccaaa ttctgaatat aggtaagtta 2040 acgtcgctac aacacattta tgtcttttct gtacagaaga agcaaggcta tgagttgcga x100 cagttgaagg acttgaatga gcttggtggc agtttaagag tggaaaatct tgagaatgtc 2160 attggaaagg atgaagccgt agagtcgaag ctatatctga aaagtcgcct taaagagttg aZZO
gtacttgagt ggagttccga gaatattctg catttggatg ttctagaggg tctgcgaccg aa80 ccaccccaac tgagtaagct cacaatcaaa ggttacagst ctgatacata tcctgggtgg 2340 ttactagagc gatcctattt tgagaatttg gaaagttttg agcttagtaa ttgcagtttg 2400 ctagaaggcc taccaccaga tacagagctc cttcggaatt gctctsggtt gtgtataaac 2460 attgttccaa atttgaagga actatctaat ctttcagcag gccttacaga tttatcaatt 2520 gattgttgcc cactgcttat gtttatcacc aacaatgagc taggacagca tgacttgagg 2580 gaaaatataa taatgaaggc agacgacctg gcatctaaac ttgcattgat gtgggaggtg 2640 gattcaggaa tagaagttag gagagtactg tcgaaagact attcatctct gaagcagttg 2700 atgacattga tgatggatgs tgatatatca aagcatcttc aaattattga aagtggtctg 2760 gaggaaagag aagataaagt atggatgaaa gaaaacatca tcaaggcatg gctcttttgc asao catgagcaga ggataagatt catttstgga aggaccatgg agatgccatt ggttctaccg 2880 tcaggactct gtgaactttc tctttcttca tgcagtatta cagatgaagc tttagctatt 2940 tgccttggtg gccccacttc sctgagaact ttacaattgg aatataatat ggcattaact 3000 acacttccat cagaaaaggt gtttgagcat ttgacaaagc ttgtcaggtt ggttgtaata 3060 gattgtttgt gtctcaaatc actggggggc ttacgtgctg ctccatctct ttcctgtttt 3120 gagtgttggg attgtccttc tttagaacta atgccgttga accttgctat cagtctcagc 3180 atccgtggct gcattcttgc agctgattcg ttcattaatg gcttgccaca tctgaaatat 3240 ctttccattg atgtctgcag aagctcccca tccttatcga ttggccacct gacctccctt 3300 gaatcattat gtctaaatgg tctccctgat ctttgctttg tgaaggcttg tcttccctgc 3360 accttaagcg cctaagttta gtagatgttg caaacctcac tgccaagtgc atctcaccgt 3420 ttcgtgtcca ggaatcgctc acggttagta gctctgtatt gctcaaccac atgctaatgg 3480 ctgaagggtt tacagcccca ccaaatctta ctcttttaga ttgcaaggag ccgtcagttt 3540 catttgaaga acctgcaaat ctctcatccg tcaagcacct gaagttttca tattgtgaaa 3600 cagagtccct gccgagaaat ctaaaatctg tctcaagtct ggagagtctt tctatacaac 3660 attgccccaa cataacatct ttaccagatc tgccgtcctc cctccagcgc ataactatat 3720 gggattgtcc cgtcttgaag aagsgctgcc aagaacctga tggagaaagc tggccaaaga 3780 tttcgcacgt tcgttggaag agctttctac caagaccgca ctggattctt tag 3833 <210> 68 <211> 3852 <212> DNA
<213> Zea mat's <400> 68 atggccgact tcgcgctcgc cggcttaagg tgggcsgcat cgccgattgt caacgagctt 60 cttactaaag cttcagctta cctcagtctg gacatggtgc gtgagatcca acgactagaa 120 gccactgtcc tgccacagtt cgagctggtg attcaagcgg cccagaagag cccccacagg 180 ggcatactgg aggcatggct ccggcgtctc aaagaagcct aetatgatgc cgaggacttg 240 ttggacgagc atgagtacaa tgtccttgag ggcaaagcca agagcggaaa sagtctcctg 300 ctgggagagc atggaagctc ctccactgca actactgtca tgaagccttt tcatgctgct 360 atgagcaggg cacggaactt gctccctcaa aacagaaggt taattagcaa gatgaacgag 4Z0 ctcaaagcaa tcctgacaga agcccaacaa cttcgagatc ttcttggctt gccacstggc 480 aataccgtcg agtgcccagc tgcsgcacct accagtgttc ccacaaccac atcacttccc 540 acttccaagg tttttggtcg cgacagggat cgtgatcata tagtagattt tcttctcgac 600 aagacaacaa ctgctcaggc aacctcagct aagtactcgg gtttggccat tgttggattg 660 ggaggaatgg ggaagtccac cttagcacag tatgtctata atgacaaaag gatsgaagaa 7Z0 tgctttgata tcaggatgtg ggtgtgcatc tcacgcaaac ttgatgtgca tcgtcacaca 7B0 agggagatta tggagtctgc aaaaaaggga gagtgcccac gtgttgataa tctcgatact 840 ctccagtgca aattacgtga tatactacaa gagtcacaga aattcctgct tgtcttggat 900 gatgtttggt ttgaaaaatc tcataatgag acagagtggg agttattcct tgctccctta 960 gtctctsaac agtcagggag caaagttttg gtgacttctc gaagtaaaac acttcctgcc 1020 gctatttgtt gtgaacaaga acatgtcatt catttggaaa acatggatga tactgagttt 1080 ttggctcttt ttaaacacca tgctttctct ggagcagaaa tcasagacca actgttacgc 1140 acgaagctgg aagacactgc agaagagatt gctaaaaggc ttggacaatg tcctttggca 1x00 gcaaaagttc tgggttctag attgtgcagg saaaaggata ttgctgaatg gaaagctgct 1260 ctgaagcttg gagatttaag tgatcccttc acatctctgt tgtggagtta cgagaagtta 1320 gatccacgtc tgcagaggtg cttcttgtat tgcagcttgt ttccsaaagg tcatagatat 1380 gaacctaatg agttggttca tctctgggtg gcagaaggat ttgttggttc atgcaatttg 1440 agtaggagsa cattggaaga ggctgggatg gattacttca atgatatggt ctctggattt 1500 ttcttccaat tggtttctaa aagacattat tcatactata tcatgcacga tatccttcat 1560 gatttggcag agtcactctc tsgagaagac tgctttagat tagasgatga taatgtgaca 1620 gagataccat gcactgttcg atatatatct gtccgtgttg aaagtatgca aaagcataag 1680 gaaattatct acaagctaca tcatttacgc actgttatct gcatcgattc actaatggat 1740 aatgcaagta ttatttttga tcagatgctg tggaacttga agaagttgcg tgtattgtct 1800 ttgtcatttt acaacagcaa caagttacct aaatctgttg gtgagctgaa gcaccttcgg 1860 tatttggacc tcaccagaac atcagtgttt gaattgccta gatcattatg tgctctttgg 1920 cacttacaac tacttcagct aaacggcatg gtggagaggt tgcctaacaa agtttgcaat 1980 ttaagtaagt tacggtatct gcgagggtat aaggaccaaa ttcccaacat tggcaagctt 2040 acttctttac aacagatata tgtcttttct gtgcaaaaga agcaaggata tgagttgcga 2100 cagctaaagg acttgaatga gcttggcggc agtttacatg acaaaaatct tgagaatgtc 2160 attggaaagg atgaagcctt agcgtcgaag ctgtatctga aaagtcgcct taaagagttg 2220 acacttgagt ggcgttctga gaatggcatg gstgcaatga atattctgca tttggatgtt 2280 ctagagggtc tgcgaccgcc accccaactg sgtaagctcs caatcaaagg ttacaastct 2340 gacacatatc ctgggtggtt acttgagcga tcctatttta agaatttgga acgttttgag 2400 cttaataatt gcagtttgct agaaggctta ccaccagata cagagctcct tcagcattgt 2460 tctagactgt tgctgttgga cgttccaaaa ctaaagacat taccatgtct tccaccaagc 2520 cttacaaagt tgtcaatttg tggcctcccc ctgcttacgt ttgtcaccaa aaatcagctc 2580 gaacascatg actctaggga aaatataatg atggcagacc acctggcatc taaactttca 2640 ttgatgtggg aggtggattc aggatctagt gttaggagtg tactgtcgaa agactattca 2700 tctcttaagc agttgatgac attgatgata gatgatgata tatcaaagca gcttcaaatt 2760 attgaaactg gtctagagga aggagataaa gtatggatga aagaaaacat catcaaggca 2s2o tggctctttt gccatgagca gaggataaga ttcacttatg gaagggocat ggagctgcaa 2880 gtggttctac cattaggact ttgtaaactt tccctttcgt catgcaatat tatagatgaa 2940 gctttsgcta tttgccttgg aggcctcact tcactggcaa ctttagaatt ggaatataat 3000 atggcactaa ctacacttcc atcagaagag gtgtttcaac atttgacaaa gcttgacatg 3060 ttgattctaa gtggttgttg gtgtctcaag tcactggggg gcttacgtgt tgcttcatct 3120 ctttccattc ttcactgttg ggattgtcct tctttagagc tagcatgtgg agcagaacta 3180 atgccgttga accttgctag caatctcacc tcccgtggct gcattcttgc agctgattcg 3240 ttcattaatg gcttgccacs tctgasacat ctttccattg atgtctgcag aagctcccca 3300 tccttatcga ttggccacct gacctccctt gaatcattac atctaaatga tctttacttt 3360 gttgaaggtc tgtcttccct gcaccttaag cacctacgtt tsgtagstgt tgcaaacctc 34x0 actgccaagt gcatctcaca gtttcgtgtc caggsatcgc tcacggttag tagctctgta 3480 ttgctcaacc acatgctaat ggctgaaggg tttacagtgc cactgaatct tgatctttca 3540 tattgcaaag agccgtcggt ttcatttgas gagcctgcaa atctctcatc tgtcaagtgc 3600 ctgggatttt ggtattgcaa aacggagtcc ctaccaagaa atctaaaatc cctctcaagt 3660 ctggagagtc tttctatagg gtgttgcccc aacatagcat ctttaccags tctgccgtcc 3720 tccctccagc gcataagtat atcaggttgc ccagtcttga agaagaactg ccasgaacct 3780 gatggggaaa gctggccaaa gatttcgcac cttcctggaa gccatctact aataccaaac 3840 ttgcttcttt ag 3852 <Z10> 69 <~11> 3853 <212> DNA
<~13> Zaa ways <400> 69 atggtggact tggcgctagt tggcttaagg tgggcagcat cgccgattgt caaggagctt 60 cttactaaag cttcagctta cctcagtgtg gacatgggag gagatcgaag actacasgac 120 actgtcctgc cacagttcga gttggtgatt caagcggccc agssgagccc ccataggggc 180 aagctggaat cctggcttcg gcgtctcaaa gaagccttct atgatgccga ggacctgctg a40 gacgagcatg agtacaacgt ccttaaggcc aaggccaaga gcggasasgg tcccctgctc 300 cgagaggatg aasgctcctc cactgcaacc acttcatgaa gccttttcat tctgctatgs 360 acagggcacg caacttgctc cctgggaaca gaaggctaat tagcaagatg aacgagctca 4Z0 aagctattct gacagaagcc aagcagcttc gagatcttct tggcttgcca catggcaata 480 ctaccgagtg gccagctgca gcacctaccc atgttcccac aactacatca cttaccactt 540 ccaaggtttt cggtcgcaac agcgatcgtg atcgcatagt aaaatttctt ctcggcaaga 600 caacaactgc tgaggcaagc tcaactaagt actccggttt ggccattgtt ggattgggag 660 gaatggggaa gtctacctta gcacaatatg tctataatga caagaggatt gaagaatgct 7Z0 ttgatgtcag gatatggatc tgtatctcgc gcaaacttga tgtgcatcgt cacacaaggg 780 agatcattga gtccgcaaaa aagggggagt gcccacgtgt tgataatctc gatactctcc 840 agtgcaaatt acgtgatata ctacaagagt cacagaaatt cctgcttgtc ttggatgatg 900 tttggtttga aaaatctcat aatgagacag agtgggagtt attccttgct ccattagtct 960 ctaaacagtc agggagcasa gttttggtga cttctcgaag tgaaacactt ccggccgcta 1020 tttgttgtgs acaagaacat gtcattcatt tggaaaacat ggatgatact gagtttttgg 1080 ctctttttaa acaccatgct ttctctggag cagaaatcaa agaccaactg ttacgcstga 1140 agctgcaaga cactgcagag gagattgcta aaaggcttgg acaatgecct ttggcagcaa 1200 aagttcttgg ttcccgaatg tgcaggagaa aggatattgc tgagtggaaa gctgctctaa 1260 agcttggaga tttaagtgat cccttcacat ctttgttatg gagctscgaa aagttagatc 1320 catgtctgca aaggtgcttc ttgtattgca gcttgtttcc aaaaggtcac ggatatagsc 1380 ctgaagagtt ggttcacctc tgggtggcag aaggatttat tggttcatgc aatttgagta 1440 ggagaacgtt agaagaggtt gggatggatt acttcaatga tstggtctct gtatctttct 1500 tccaaaggta cggttggtac tatgtcatgc atgatatcct tcatgatttt gcagagtcac 1560 tctctagaga agactgcttt agattagaag atgataatgt gacagagata ccatgcactg 1620 ttcgacatct atctgttcgt gttgaaagta tgcaaaagca taaggaaatt atctacaagc 1680 tacatcattt acgcactgtt atctgcatcg attcactaat ggataatgca agtattattt 1740 ttgatcagat gctgtggaac ttgaagaagt tgcgtgtstt gtctttgtca tttcacaaca 1800 gcaacaagtt acctaaatct gttggtgagc tgaagcacct tcggtatttg gacctcaaca 1860 gaacatcagt gtttgaattg cctagatcat tatgtgctct ttggcactta caactacttc 1920 agctaaacgg catggtggag aggttgccta acsaagtttg csatttaagt aagttacggt 1980 atctgcgagg gtataaggac caaattccca acattggcaa gcttaettct ttacaacaga 2040 tatatgactt ttctgtgcaa aagaagcaag gatatgagtt gcgacagcta aaggacttga 1100 atgagcttgg cggcagttta catgtccaaa ttcttgagaa tgtcattgga aaggatgaag x160 ccttagcgtc gaagctatat ctaaaaagtc gccttaaaga gttgatactt gagtggagtt 2Za0 ctgagaatgg catggatgca atgaatattc tgcatttgga tgttctagag ggtctgcgac aa8o cgccacccca actgagtaag ctcacaatcg aaggttscag atctgataca tatcctgggt 2340 ggttactaga gcgatcctat tttgagaatt tggaaagttt tgagcttagt aattgcagtt 2400 tgctagaagg cctaccacca gatacagagc tcgttcggaa ttgctctagg ttgcgtataa 2460 acattgttcc aaatttgsag gaactatcta atcttccagt aggccttaca gatttatcaa 25x0 ttgattattg cccactgctt atgtttatca ccaacaatga gctaggacag catgacttga x580 gggaaaatat aataatgaag gcagacgacc tggcatctaa acttgcattg acgtgggagg x640 tggattcagg saaagttagg agagtactgt cgaaagacta ttcatctctg aagcaattga x700 tgacattgat gatggatgat gatatatcaa agcatcttca aattattgaa agtggtctgg 2760 aagaaagaga agataaagta tggatgaaag aaaacatcat caaggcatgg ctcttttgcc Z8a0 atgagcagag gataagattc atttatggaa ggaccatgga catgccattg gttctaccgt 2880 caagactctg tgaactttct ctttcttcat gcagtattac agatgaagct ttagctattt 2940 gccttggtgg cctcacttca ctgagcaatt taaaattgaa atataatatg gcattaacta 3000 cacttccatc agaaaaggtg tttgagcatt tgacaaagct tgacacgttg gttgtaacag 3060 gttgtttgtg tctcaaatca atggggggct tacgtgctgc tccatctctt tccttttttt 31x0 actgttcgga ttgtcctttt ttagagctag cacgtggagc agaactaatg ccgttgaacc 3180 ttgatggaga cctccacatc cgtggctgca ttcttgcagc tggatcgttc attaatggct 3240 tgccacatct gaaacatctt tcctttgatg tctgcagaag ctccccatcc ttatcgattg 3300 gccacctgac ctcccttgaa tcattacgtc taaatggtct ccctgatctt tactctgttg 3360 aaggcttgtc tgccctgcac cttaagtacc taactctaca agatgttgca aacctcactg 3420 tcaagtgcat ctcacagttt cgtgtccagg aatcgctcac ggttagtacc tccgtattgc 3480 tcaaccacat gctcatggct gaaggattts cagtcccacc gaatcttgat ctttcatatt 3540 gcaaagaacc gtcggtttca tttgaagagc ctgcaaatct ctcatctgtc aagtgcctgg 3600 gattttggta ttgcaaaacg gagtccctac caagaaatct aaaatccctc tcaagtctgg 3660 agagtctttc tatagggtgt tgccccaaaa tagcatcttt aacsgatatg ccgtcctccc 37x0 tccagcgcat aagtatagta aattgccccg tcttgaagaa gaactgccaa gaacctgatg 3780 gggaaagctg gccaaagatt tcgcaccttc gccggacgca catcaactgc taataccaaa 3840 cttgcttctt tag 3853 <a10> 70 <211> 1896 <212> DNA
<213> Zea ways <400> 70 atggccgact tggcgctcgc cggcttaagg tgggcagcat cgccgattgt caacgagctt 60 cttactaaag cttcagctta cctcagtgtg gacatggtgc gtgagatcca, acgactagaa 120 gccactgtcc tgccacagtt cgagctggtg attcaagcgg cccagaagag cccccacagg 180 ggcatactgg aggcatggct ccggcgtctc aaagaagcct actatgatgc cgaggacttg 240 ~ ttggacgagc atgagtacaa tgtccttgag ggcaaggcca agagcgaaaa aagsctcctg 300 ctgggagagc atggaagctc ctccactgca actactgtca tgaagccttt tcatgctgct 360 atgagcaggg cacggaactt gctccctcaa aacagaaggc taattagcaa gatgaacgag 4Z0 ctcaaagcaa tcctgacaga agcccaacaa cttcgagatc ttcttggttt gccacatggc 480 aataccgtcg agtggccagc tgcagcacct accagtgttc ccacaaccac atcacttacc 540 scttccaagg tttttggtcg cgacagggat cgtgatcgta tagtagattt tcttctcggc 600 aagacaacaa ctgctgaggc aagctcagct aagtactcgg gtttggccat tgttggattg 660 ggaggaatgg ggaagtccac cttagcacag tatgtctata atQacaaaag gatsgaagas 7a0 tgctttgata tcaggatgtg ggtgtgcatc tcacgcaaaa ttgatgtgca tcgtcaaaca 780 agggagatta tagsgtctgc aaaaaaggga gagtgcccac gtgttgstaa tctcgatact 840 ctccagtgca aattacgtga tatactacaa gagtcacaga aattcttgct tgtcttggat 900 gatgtttggt ttgaaaaatc tcataatgag acagagtggg agttattcct tgctccatta 960 gtctctaaac agtcagggag caaagttttg gtgacttctc gaagtaaaac acttcctgcc 1020 gctatttgtt gtgaacaaga acatgtcatt catttgaaaa acatggatga tactgagttt 1080 ttggctcttt ttaaacacca tgctttctct ggagcagaaa tcaaagacca actattacgc 1140 acgaagctgg aagacactgc agtggagatt gctaaaaggc ttggacaatg tcctttggca 1x00 gcaaaagttc tgggttctcg attgtgcagg aaaaaggats ttgctgaatg gaaagctgct 1260 ctaaagcttg gagatttaag tgstcccttc acatctctgt tgtggagtta cgagaagtta 13x0 gatccacgtc tgcagaggtg cttcttgtat tgcagcttgt ttccaaaagg tcatagatat 1380 gatcctaatc agttggtcat agatagatat ttatgcaaag attgcccttt catgaaagta 1440 gagaggtgtg gaacaaaatt ttgatatggg ggaactgttc tttcctggga ggaaaccaga 1500 catcggagtc gttgtatgat tggtggagga atctaagagg tctctgtaat aggcaatcta 1560 gaaaaaaatt cgatgggctt ctgatctact tttggtggag cttatggtta gaaagaaaca 160 acaggatctt csgaaatcag cagaagastt cagatcaggt cgcttattta gtgagagagc 1680 ttgttggcgc tctagtgggt tagttttggg cctagtggct ttggagttgt ttttttaatc 1740 ttagtagaca gtcgagtttt gctcttcttc tttcttcttc cccctctctc tttttcgtct 1800 tcgtgtgtgt cttgtaagtg ttttctcctt ctctaatata ttggaccggc aaatcttttg 1860 cecgtccctt tcaaaa 1876

Claims (73)

CLAIMS:

1. An isolated nucleic acid molecule comprising an Kp1-D nucleotide sequence or Rpg1 nucleotide sequence or hrp1 nucleotide sequence or a variant thereof which encodes a protein or derivative thereof, which confers, enhances, or otherwise facilitates resistance to a pathogen in a plant, wherein the Rp1-D nucleotide sequence or hrp1 nucleotide sequence or variant thereof comprises a nucleotide sequence that is at least 60% identical to any one of SEQ ID NO:<400>1 or SEQ ID NO:<400>3 or SEQ ID NOS:<400>64 to <400>70 or a complementary nucleotide sequence thereto.

2. The isolated nucleic acid molecule according to claim 1, wherein the variant is derived from a monocotyledonous plant.

3. The isolated nucleic acid molecule according to claim 2, wherein the monocotyledonous plant is selected from the list comprising Zea mays, pearl millet, sugar cane, wheat, barley, rye, rice, oats, triticale, sorghum and ryegrass.

4. The isolated nucleic acid molecule according to claim 3, wherein the monocotyledonous plant is Zea mays or barley.

5. The isolated nucleic acid molecule according to claim 1, wherein the Rp1-D
nucleotide sequence or variant thereof comprises the nucleotide sequence set forth in any one of SEQ ID NO:<400>1 or SEQ ID NO:<400>3 or SEQ ID NOS:<400>64 to <400>70 or a derivative of said nucleotide sequence comprising at least about contiguous nucleotides of any one of SEQ ID NOS: <400>1 and/or <400>3 and/or SEQ
ID NOS:<400>64 to <400>70.

6. The isolated nucleic acid molecule according to claim 1, wherein the Rpg1 nucleotide sequence or variant thereof comprises a nucleotide sequence that is at least 60% identical to SEQ ID NO:<400>5 or a complementary nucleotide sequence thereto.

7. The isolated nucleic acid molecule according to claim 6, wherein the Rpg1 variant is derived from a monocotyledonous plant.

8. The isolated nucleic acid molecule according to claim 7, wherein the monocotyledonous plant is selected from the list comprising Zea mays, pearl millet, sugar cane, wheat, barley, rye, rice, oats, triticale, sorghum and ryegrass.

9. The isolated nucleic acid molecule according to claim 8, wherein the monocotyledonous plant is Zea mays or barley.

10. The isolated nucleic acid molecule according to claim 1, wherein the Rpg1 nucleotide sequence or variant thereof comprises the nucleotide sequence set forth in SEQ ID NO:<400>5 or a derivative of said nucleotide sequence comprising at least about 30 contiguous nucleotides of SEQ ID NO: <400>5.

11. The isolated nucleic acid molecule according to claim 1, wherein the Rpg1 nucleotide sequence or variant thereof comprises a nucleotide sequence that is at least 60% identical to SEQ ID NO:<400>7 or a complementary nucleotide sequence thereto.

12. The isolated nucleic acid molecule according to claim 11, wherein the Rpg1 variant is derived from a monocotyledonous plant.

13. The isolated nucleic acid molecule according to claim 12, wherein the monocotyledonous plant is selected from the list comprising Zea mays, pearl millet, sugar cane, wheat, barley, rye, rice, oats, triticale, sorghum and ryegrass.

14. The isolated nucleic acid molecule according to claim 13, wherein the monocotyledonous plant is Zea mays or barley.

15. The isolated nucleic acid molecule according to claim 14, wherein the Rpg1 nucleotide sequence or variant thereof comprises the nucleotide sequence set forth in SEQ ID NO:<400>7 or a derivative of said nucleotide sequence comprising at least about 30 contiguous nucleotides of SEQ ID NO: <400>7.

16. An isolated nucleic acid molecule which encodes a protein or derivative thereof, which confers, enhances, or otherwise facilitates resistance to a pathogen in a plant, wherein said nucleic acid molecule comprises a nucleotide sequence which hybridises under at least low stringency conditions to at least about 25-30 contiguous nucleotides of SEQ ID NOS: <400>1 and/or <400>3 and/or <400>5 and/or <400>7 and/or any one of SEQ ID NOS:<400>64 to <400>70 or a complementary sequence thereto.

17. The isolated nucleic acid molecule according to claim 16, derived from a monocotyledonous plant.

18. The isolated nucleic acid molecule according to claim 17, wherein the monocotyledonous plant is selected from the list comprising Zea mays, pearl millet, sugar cane, wheat, barley, rye, rice, oats, triticale, sorghum and ryegrass.

19. The isolated nucleic acid molecule according to claim 18, wherein the monocotyledonous plant is Zea mays or barley.

20. The isolated nucleic acid molecule according to claim 16 comprising the nucleotide sequence set forth in SEQ ID NOS: <400>1 and/or <400>3 and/or any one of SEQ ID NOS:<400>64 to <400>70 or a complementary sequence thereto.

21. The isolated nucleic acid molecule according to claim 16 comprising the nucleotide sequence set forth in SEQ ID NO: <400>5 or a complementary sequence thereto.

22. The isolated nucleic acid molecule according to claim 16 comprising the nucleotide sequence set forth in SEQ ID NO: <400>7 or a complementary sequence thereto.

23. An isolated nucleic acid molecule which encodes a pathogen-resistance protein or derivative thereof, which confers, enhances, or otherwise facilitates resistance to a pathogen in a plant, wherein said pathogen-resistance protein or derivative comprises an amino acid sequence set forth in any one of SEQ ID NOS: <400>2, <400>4, <400>6 or <400>8 or a homologue, analogue or derivative of said amino acid sequences that is at least about 60% identical thereto.

24. The isolated nucleic acid molecule according to claim 23, derived from a monocotyledonous plant.

25. The isolated nucleic acid molecule according to claim 24, wherein the monocotyledonous plant is selected from the list comprising Zea mays, pearl millet, sugar cane, wheat, barley, rye, rice, oats, triticale, sorghum and ryegrass.

26. The isolated nucleic acid molecule according to claim 25, wherein the monocotyledonous plant is Zea mays or barley.

27. The isolated nucleic acid molecule according to claim 23, wherein the pathogen-resistance protein or derivative comprises the amino acid sequence set forth in SEQ
ID NOS: <400>2 or <400>4 or at least 10 contiguous amino acids thereof.

28. The isolated nucleic acid molecule according to claim 23, wherein the pathogen-resistance protein or derivative comprises the amino acid sequence set forth in SEQ
ID NO: <400>6 or at least 10 contiguous amino acids thereof.

29. The isolated nucleic acid molecule according to claim 23, wherein the pathogen-resistance protein or derivative comprises the amino acid sequence set forth in SEQ
ID NO: <400>8 or at least 10 contiguous amino acids thereof.

30. The isolated nucleic acid molecule according to claim 23, wherein the pathogen-resistance protein or derivative further includes one or more of the amino acid sequences set forth in any one of SEQ ID NOS: <400>17 to <400>33.

31. A method of identifying a pathogen resistance genetic sequence or pathogen resistance-like genetic sequence in a plant, said method comprising contacting genomic DNA, mRNA, cDNA or a fragment or a source thereof, with a hybridisation-effective amount of a probe comprising at least about 25-30 contiguous nucleotides of any one of SEQ ID NOS: <400>1 or <400>3 or <400>5 or <400>7 or any one of SEQ
ID NOS:<400>64 to <400>70 or a complementary nucleotide sequence thereto for a time and under conditions sufficient for hybridisation to occur and then detecting said hybridisation.

32. The method according to claim 31 wherein the probe is labelled with a reporter molecule.

33. The method according to claim 31 wherein the plant is a broadacre crop plant and/or a monocotyledonous plant.

34. The method according to claim 33, wherein the monocotyledonous plant is maize, barley, rye, oat, wheat, sorghum, triticale, ryegrass or rice or a wild variety and/or hybrid or derivative and/or ancestral progenitor of same.

35. A method of identifying a pathogen resistance-genetic sequence or pathogen resistance-like genetic sequence in a plant, said method comprising hybridising one or more nucleic acid primer molecules of at least about 25-30 nucleotides in length of any one of SEQ ID NOS: <400>1 or <400>3 or <400>5 or <400>7 or any one of SEQ
ID NOS:<400>64 to <400>70 or a complementary nucleotide sequence thereto with genomic DNA, mRNA, cDNA or a fragment or a source thereof, and amplifying nucleic acid therefrom in a polymerase chain reaction.

36. The method according to claim 35 wherein the primer comprises a nucleotide sequence set forth in any one of SEQ ID NOS:<400>34 to <400>63.

37. The method according to claim 35 wherein the plant is a broadacre crop plant and/or a monocotyledonous plant.

38. The method according to claim 37, wherein the monocotyledonous plant is maize, barley, rye, oat, wheat, sorghum, triticale, ryegrass or rice or a wild variety and/or hybrid or derivative and/or ancestral progenitor of same.

39. A genetic construct that comprises the isolated nucleic acid molecule according to claim 1 or the protein-encoding region thereof in operable connection with a promoter sequence.

40. The genetic construct according to claim 39, wherein the promoter sequence is operable in a plant cell, tissue, organ or whole plant.

41. The genetic construct according to claim 40, wherein the promoter sequence comprises nucleotides 1 to about 1200 of SEQ ID NO: <400>1 or nucleotides 1 to about 3765 of SEQ ID NO: <400>5, or nucleotides 1 to about 715 of SEQ ID NO:
<400>7 or a homologue, analogue or derivative thereof which is capable of conferring expression on a structural gene in a plant cell.

42. The genetic construct according to claim40, wherein the promoter sequence comprises the CaMV 35S promoter, NOS promoter, OCS promoter, A. thaliana SSU
promoter or napin promoter sequence.

43. An isolated promoter sequence that is capable of conferring expression on a structural gene in a plant cell comprising nucleotides 1 to about 1200 of SEQ
ID NO:
<400>1.

44. An isolated promoter sequence that is capable of conferring expression on a structural gene in a plant cell comprising nucleotides 1 to about 3765 of SEQ
ID NO:
<400>5.

45. An isolated promoter sequence that is capable of conferring expression on a structural gene in a plant cell comprising nucleotides 1 to about 715 of SEQ
ID NO:
<400>7.

46. A pathogen resistance protein substantially free of conspecific proteins and having at least about 60% amino acid sequence identity to any one or more of SEQ ID
NOS:<400>2, <400>4, <400>6 or <400>8, wherein said pathogen resistance protein confers, enhances, or otherwise facilitates resistance to a pathogen in a plant.

47. The pathogen resistance protein according to claim 46 comprising the amino acid sequence set forth in SEQ ID NOS: <400>2 or <400>4 or at least 10 contiguous amino acids thereof.

48. The pathogen resistance protein according to claim 46 comprising the amino acid sequence set forth in SEQ ID NO: <400>6 or at least 10 contiguous amino acids thereof.

49. The pathogen resistance protein according to claim 46 comprising the amino acid sequence set forth in SEQ ID NO: <400>8 or at least 10 contiguous amino acids thereof.

50. The pathogen resistance protein according to claim 46 further including one or more of the amino acid sequences set forth in any one of SEQ ID NOS: <400>17 to <400>33.

51. An antibody molecule that binds to the pathogen resistance protein according to claim 46 or an epitope thereof.

52. A method of producing a plant having improved resistance to a plant pathogen comprising expressing a pathogen resistance protein which comprises an amino acid sequence having at least about 60% amino acid sequence identity to any one or more of SEQ ID NOS:<400>2, <400>4, <400>6 or <400>8 in said plant or a cell, tissue or organ thereof.

53. The method according to claim 52, further comprising introducing a nucleic acid molecule encoding the pathogen resistance protein into a plant cell, tissue, organ or whole plant in a format suitable for expression to occur.

54. The method according to claim 52, further comprising introducing a nucleic acid molecule encoding the pathogen resistance protein into a plant cell, tissue or organ in a format suitable for expression to occur and regenerating a whole plant therefrom.

55. The method according to claims 53 or 54, wherein the nucleic acid molecule is in operable connection with a promoter sequence capable of being expressed in a plant cell, tissue, organ or whole plant.

56. The method according to claim 52, wherein the pathogen resistance protein comprises an amino acid sequence set forth in SEQ ID NOS:<400>2 or <400>4.

57. The method according to claim 52, wherein the pathogen resistance protein comprises an amino acid sequence set forth in SEQ ID NO:<400>6.

58. The method according to claim 52, wherein the pathogen resistance protein comprises an amino acid sequence set forth in SEQ ID NO:<400>8.

59. The method according to claim 52 wherein the plant pathogen is a fungus.

60. The method according to claim 52 wherein the plant pathogen is a rust.

61. The method according to claim 60 wherein the rust is Puccinia sp.

62. The method according to claim 60 wherein the rust is barley stem rust.

63. The method according to claim 60 wherein the rust is Pyricularia oryzae.

64. The method according to claim 60 wherein the rust is Xanthomonas oryzae.

65. The method according to claim 52 wherein the plant pathogen is a virus.

66. The method according to claim 65 wherein the virus is wheat streak mosaic virus.

67. The method according to claim 52 wherein the plant pathogen is a bacterium.

68. The method according to claim 52 wherein the plant pathogen is a nematode.

69. A plant produced by the method comprising:
(i) introducing a nucleic acid molecule comprising the nucleotide sequence set forth in any one of SEQ ID NOS: <400>1 or <400>3 or <400>5 or <400>7 or any one of SEQ ID NOS:<400>64 to <400>70 or a protein-encoding fragment thereof into a plant cell, tissue or organ in a format suitable for expression to occur; and (ii) regenerating a whole plant therefrom.

70. The plant according to claim 69 capable of expressing the pathogen resistance protein or a biologically active fragment thereof encoded by the introduced nucleic acid molecule.

71. The plant according to claim 70 having improved resistance to a plant virus, fungus, bacteria, nematode or rust pathogen.

72. The plant according to claim 71 wherein the rust is Puccinia sp.

73. The plant according to claim 72 wherein the rust is barley stem rust.