Abstract:
A class of wheat SEBII genes, called SBEII-1, can be used to influence properties of starch produced by a plant, including the gelatinisation and/or pasting temperature of the starch. Such a produced starch is useful, for example, in certain industrial applications and the preparation and/or processing of foodstuffs such as bakery products. One aspect of the present invention provides a nucleotide sequence encoding the amino acid sequence shown in FIG.  10  (SEQ ID No: 2).

Description:
This application is a §371 national stage filing of PCT/GB99/03011, filed Sep. 9, 1999 (published in English on Mar. 23, 2000 as WO 00/15810) and claiming priority to EP 98307337.0 filed Sep. 10, 1998. 
    
    
     FIELD OF THE INVENTION 
     This invention relates generally to plant starch compositions, and concerns novel nucleotide sequences; polypeptides encoded thereby; vectors and host cells and host organisms comprising one or more of the novel sequences; a method of altering one or more characteristics of a plant; a plant having altered characteristics; starch obtained from such plants; and uses of the starch. 
     BACKGROUND TO THE INVENTION 
     The majority of developments in cereal science in the recent past have concentrated primarily on the functionality of the gluten protein sub-units and their role in bakery systems. This has been greatly facilitated by the abundance of natural variation between cultivators for the gluten protein sub-unit components. 
     In contrast, although flour from commercially grown wheat varieties contains approximately 75-85% starch, the role of starch from a breeding perspective has been overlooked; this is largely due to the difficulty of measuring differences in starch structure. Of the limited amount of work that has been carried out however, there appears to be a lack of natural variation between different wheat cultivars. With the advent of recombinant DNA and gene transfer technologies it is now possible to create new variation in planta, therefore directly modifying starch composition in wheat becomes a realistic target. 
     Starch is the major form of carbon reserve in plants, constituting 50% or more of the dry weight of many storage organs, e.g. tubers, seeds of cereals. Starch is used in numerous food and industrial applications. In many cases, however, it is necessary to modify the native starches, via chemical or physical means, in order to produce distinct properties to suit particular applications. It would be highly desirable to be able to produce starches with the required properties directly in the plant, thereby removing the need for additional modification. To achieve this via genetic engineering requires knowledge of the metabolic pathway of starch biosynthesis. This includes characterisation of genes and encoded gene products which catalyse the synthesis of starch. Knowledge about the regulation of starch biosynthesis raises the possibility of “re-programming” biosynthetic pathways to create starches with novel properties that could have new commercial applications. 
     The most significant property of starch derives from the ability of the native granular form to lose its order and to swell and absorb water upon suitable treatment, thereby conferring viscosity and texture, in a process known as gelatinisation. Gelatinisation has been defined (W A Atwell et al, 1988) as “. . . the collapse (disruption) of molecular orders within the starch granule manifested in irreversible changes in properties such as granular swelling, native crystallite melting, loss of birefringence, and starch solubilisation. The point of initial gelatinisation and the range over which it occurs is governed by starch concentration, method of observation, granule type, and heterogeneities within the granule population under observation”. 
     14 molecules of water per molecule of anhydrous glucose, i.e. a minimum of 75 % water, are necessary for full starch gelatinisation (Donovan, 1979). Starch gelatinisation is usually caused by heat, but can be caused by physical damage and some chaotropic agents, mainly dimethylsulphoxide (DMSO), urea, calcium chloride, strong base and acid. 
     The various events taking place during gelatinisation can be followed by various methods, including birefringence, X-ray diffraction, differential scanning calorimetry (DSC),  13 C NMR. Swelling can be monitored by various methods, particularly rheology. 
     Differential scanning calorimetry (DSC) is a destructive method which records an endothermic event on heating of granules, generally thought to measure the temperature and the endothermic energy (delta H) required for the melting of the native crystallites. Starch gelatinisation temperature is independent of water content above 75% water (described as excess water), but increases when water is limited (Donovan, 1979). 
     The rate and extent of starch granule swelling upon heating dictate the type of viscosity development of aqueous starch suspensions on heating. Swelling behaviour is therefore of utmost technological importance. Viscosity increase on heating can be conveniently measured by a Brabender amylograph (Brabender is a Trade Mark) (Kennedy and Cabalda, 1991) or using a Rapid Visco analyser (Rapid Visco is a Trade Mark from Newport Scientific, Australia). FIG. 1 is a typical viscoamylgraph profile for wheat starch, produced in this way, showing changes in starch during and after cooking. As starch granules swell on uptake of water, in a process known as pasting, their phase volume increases, causing an increase in viscosity. The onset of pasting is indicated at A in FIG.  1 . Peak viscosity, indicated at B in FIG. 1, is achieved when maximum phase volume is reached. Shear will then disrupt/cause fragmentation of the swollen granules, causing the viscosity to decrease. Complete dispersion is indicated at C in FIG.  1 . This has been confirmed by an oscillatory rheology study of starch pastes at various stages of the viscosity profile (Svegmark and Hermansson, 1990). The viscosity onset temperature and peak viscosity are indicative of the initiation and extent of swelling, respectively. On cooling, leached amylose forms a network in a process involving reassociation of molecules, or retrogradation, causing an increase in viscosity as indicated at D in FIG.  1 . Retrogradation (or set-back) viscosity is therefore indicative of the amount of amylose leached out of the granules. 
     The properties of wheat starch are useful in a large number of applications and also non-food (paper, textiles, adhesives etc.) applications. However, for many applications, properties are not optimum and various chemical and physical modifications well known in the art are undertaken in order to improve useful properties. Two types of property manipulation which would be of use are: the controlled alteration of gelatinisation and pasting temperatures; and starches which suffer less granular fragmentation during pasting than conventional starches. 
     Currently the only ways of manipulating the gelatinisation and pasting temperatures of starch are by the inclusion of additives such as sugars, polyhydroxy compounds of salts or by extensive physical or chemical pre-treatments. The reduction of granule fragmentation during pasting can be achieved either by extensive physical pre-treatments or by chemical cross-linking. Such processes are inconvenient and inefficient. It is therefore desirable to obtain plants which produce starch which intrinsically possesses such advantageous properties. 
     Starch consists of two main glucose polysaccharides: amylose and amylopectin. Amylose is a generally linear polymer comprising α-1,4 linked glucose units, while amylopectin is a highly branched polymer consisting of an α-1,4 linked glucan backbone with α-1,6 linked glucan branches. In wheat endosperm amylopectin constitutes approximately 70% of the total starch content, with the balance being amylose. Amylopectin is synthesised through the concerted action of several enzymes, including soluble starch synthase(s) (SSS), starch branching enzyme(s) (SBE), starch de-branching enzyme(s) (DBE). The physical properties of starch are strongly affected by the relative abundance of amylose and amylopectin, therefore SSSs, SBEs and DBEs play a key role in determining both starch quantity and quality. As such, one approach to manipulating starch structure would be to modify the expression of the enzymes involved in starch biosynthesis in the endosperm using a transgenic approach. 
     SBE catalyses the formation of the α-1,6 linkages, creating branch points in the growing starch molecule, via hydrolysis of an α-1,4 linkage followed by reattachment of the released α-1,4-glucan chain to the same or another glucosyl chain. This reaction also provides a new non-reducing end for further elongation of the original α-1,4-glucan chain. 
     Multiple isoforms of starch branching enzyme have been described, biochemically, from a number of species including maize (Boyer and Preiss, 1978), rice (Nakamura et al., 1992), pea (Smith, 1988), potato (Khoshnoodi et al., 1993) and wheat (Morell et al., 1997). More recently, genomic and cDNA sequences for SBE have been characterised from several species including maize (Baba et al., 1991; Fisher et al., 1993; Gao et al. 1997) pea (Burton et al., 1995), potato (Kossmann et al., 1991), rice (Nakamura and Yamanouchi, 1992; Mizuno et al., 1993), Arabidopsis (Fisher et al., 1996), cassava (Salehuzzaman et al., 1992), and wheat (Rapellin et al., 1997, Nair et al., 1997, Rahman et al., 1997). Sequence alignment of these SBEs revealed a high degree of sequence conservation at the amino acid level and that the SBEs may be grouped into two distinct families, generally known as SBEI and SBEII. Further, analysis indicates that within a species there is generally of the order of 50% homology between the two families, SBEI and SBEII, while there is often greater homology within the two families between species. 
     Maize is unusual in that the maize SBEII family is thought to comprise two different members, known as SBEIIa and SBEIIb. There has been controversy over whether the SBEIIa and IIb enzymes are in fact a) encoded by genes at two different loci, and b) whether the genes represent different alleles at a single locus. Fisher et al (1996) and Gao et al (1997) have provided evidence that SBEIIa and SBEIIb are encoded by independent genes. However, there is no conclusive evidence that both isoforms exist together in any one maize genotype. The DNA clones for the two published gene sequences were purified from different genotypes of maize and it is thus possible that they represent different alleles of a single locus. In summary, in maize, three distinct SBE genes have been characterised to date (Baba et al., 1991; Fisher et al., 1993; Gao et al., 1997). SBEI is distinct from SBEIIa and SBEIIb in amino acid composition, substrate specificity, kinetic properties, and immunological reactivities, whereas SBEIIa and SBEIIb are similar in these respects (Guan and Preiss, 1993; Preiss 1991; Takeda et al., 1993). At the amino acid level the sequence exhibits approximately 50% homology with the SBEIIa and SBEIIb sequences, whereas SBEIIa and SBEIIb exhibit approximately 80% homology to each other. 
     Prior to the present invention, maize was unique in having SBEIIa- and SBEIIb-type enzymes. Although Arabidopsis has two SBEII family members, the sub-division in Arabidopsis does not appear to conform to that seen in maize: the Arabidopsis sub-family members do not obviously fall into the IIa and IIb categories as do the maize sequences. Both of the Arabidopsis SBEII genes have similar levels of homology to both the maize SBEII genes, SBEIIa and SBEIIb, but the similarities are not sufficient to be able to place the Arabidopsis genes into the same SBEIIa and SBEIIb categories as for maize. Indeed, the data, if anything, suggests that the Arabidopsis SBEII genes do not fall into the maize IIa and IIb categories. For barley, two forms of SBEII had been partly characterised. Although these have been called SBEIIa and SBEIIb, only a very limited amount of sequence information had been published (Sun et al, 1995) and it was not possible to infer or conclude that these forms correspond to the IIa and IIb categories of maize. In fact, based on the available barley sequence information both of the barley SBEII sequences (SBEIIa and SBEIIb) would appear to show greater homology to maize SBEIIa than to maize SBEIIb. 
     For all other plant species for which SBEII sequences have been identified and published, including potato, pea, rice, cassava, wheat and barley, no subdivision of the SBEII family comparable to the SBEIIa and SBEIIb division of maize has been made. 
     Studies of purified SBEI and SBEII demonstrate that these isoforms differ in their specificity for a substrate with respect to both chain length and degree of branching. In maize, SBEI and SBEII show distinct branching activities in vitro, with SBEI showing a higher rate of branching of an amylose substrate when compared to SBEII whereas both SBEIIa and IIb show higher rates of branching than SBEI when acting upon an amylopectin substrate (Guan and Preiss, 1993). Furthermore, maize SBEI preferentially transfers longer glucan chains (average chain length=24) than SBEII (average chain length=21(IIa) and 22(IIb)) (Takeda et al., 1993). A similar observation has been reported for SBEI and SBEII isoforms from wheat and pea (Morell et al., 1997; Smith, 1988). Mutational studies in maize, rice and pea demonstrate that high amylose mutants in each case are deficient in the branching enzyme activity analogous to maize SBEII (Martin and Smith, 1995; Morell et al., 1995). However, the linkage between the biochemical observations and the genetic evidence suggesting the differences in the roles remains unclear. 
     The present invention is based on the unexpected discovery of a novel class of SBEII genes in wheat, referred to herein as SBEII-1. The novel SBEII-1 gene sequence has strong homology with the maize SBEIIb gene. The wheat SBEII-1 genes are thought to be functionally equivalent to the maize SBEIIb gene, and on this basis it is believed that manipulation of the wheat SBEII-1 gene is likely to influence starch properties including starch gelatinisation temperature, in a manner analogous to manipulation of the maize SBEIIb gene as described in WO 97/22703. 
     In summary, although two different SBEII gene sequences are known from maize, Arabidopsis and barley, as discussed above, prior to the present invention there was no reason to expect that wheat would show a similar sub-division of SBEII genes as is seen for maize. The two Arabidopsis SBEII genes show a different sub-division, and prior to the present invention there was insufficient evidence to determine whether the two barley SBEII sequences belonged to the maize-type sub-division. That is, prior to the present invention there was no reason to expect that wheat would have two similar SBEII members comparable to those of maize. Subsequent to the present invention Sun et al (1998) have presented data which indicates that the barley sequences do indeed sub-divide in a similar manner to the maize SBEIIa and IIb sequences and the wheat SBEII-2 and SBEII-1 sequences discussed in this document. 
     The present inventors have used the high degree of sequence conservation between several SBE gene sequences to design oligonucleotide primers to motifs which are specific to either SBEI or SBEII families and have used these primers to amplify cDNA sequences from developing endosperm of wheat. 
     When this work was started, a single partial length wheat SBE cDNA clone had been reported (Mousley, 1994). Multiple sequence alignment of this wheat SBE sequence with other published SBE sequences from a number of plant species revealed a number of motifs which were highly conserved. Oligonucleotide primers designed to be complementary to these motifs were used to clone 3′ partial length cDNA clones of wheat SBEII. Alignment of the cDNA clone sequences indicated that the clones could be divided into two classes, which the inventors have designated SBEII-1 and SBEII-2, which showed greater than 90% similarity to members within a class but only 60% similarity between classes. Significantly, comparison between representative sequences from each class with previously identified wheat SBEII clones, pWBE6 (Mousley, 1994) and SBEII (Nair et al., 1997), showed that each appear to be homologues of the SBEII-2 class. The cloning of a wheat SBEII-1 cDNA is novel. 
     SUMMARY OF THE INVENTION 
     In one aspect the invention provides a nucleotide sequence encoding substantially the amino acid sequence shown in FIG. 10 (SEQ ID No: 2) or a functional equivalent of said nucleotide sequence. 
     The term functional equivalent is used in this context to encompass those sequences which differ in their nucleotide composition to that shown in FIG. 10 (SEQ ID No: 1) but which, by virtue of the degeneracy of the genetic code, encode polypeptides having identical or substantially identical amino acid sequences. It is intended that the term should generally apply to sequences which are sufficiently homologous to the sequence of the invention that they can hybridise to the complement thereof under stringent hybridisation conditions (eg as described by Sambrook et al 1989, ie washing with 0.1×SSC, 0.5% SDS at 68° C.); such equivalents will preferably possess at least 86%, more preferably at least 90%, and most preferably at least 95%, sequence homology (ie sequence similarity) with the sequence of the invention. Sequence homology is suitably determined using the ‘MEGALIGN’ program of the software package DNAStar (MEGALIGN and DNAStar are Trade Marks). It will be apparent to those skilled in the art that the nucleotide sequence of the invention may also find useful application when present as an “antisense” sequence. Accordingly, functionally equivalent sequences will also include those sequences which can hybridise, under stringent hybridisation conditions, to the sequence of the invention (rather than the complement thereof). Such “antisense” equivalents will preferably possess at least 86%, more preferably at least 90%, and most preferably 95% sequence homology with the complement of the sequence of the invention. 
     In another aspect, the invention provides a nucleotide sequence comprising substantially the sequence of B2 shown in FIG. 3 (SEQ ID No: 3), or a functional equivalent thereof. 
     In a further aspect, the invention provides a nucleotide sequence comprising substantially the sequence of B4 shown in FIG. 3 (SEQ ID No: 4), or a functional equivalent thereof. 
     Another aspect of the invention provides a nucleotide sequence comprising substantially the sequence of B10 shown in FIG. 3 (SEQ ID No: 5), or a functional equivalent thereof. 
     Yet a further aspect of the invention provides a nucleotide sequence comprising substantially the sequence of B1 shown in FIG. 3 (SEQ ID No: 6), or a functional equivalent thereof. 
     In another aspect the invention provides a nucleotide sequence encoding substantially the amino acid sequence of B6 shown in FIG. 4 (SEQ ID No: 7), or a functional equivalent thereof. 
     The term functional equivalent in this context has the same general meaning as discussed above, although equivalents for B2, B4, B10 and B6 will preferably possess at least 90%, more preferably at least 95%, sequence homology with the relevant sequence of the invention, while equivalents for B1 will preferably possess at least 97% sequence homology with the sequence of the invention. 
     The sequences of the invention are part of novel wheat SBEII genes, with B1 being a novel subclass of the known class of SBEII genes, referred to herein as SBEII-2, with the novel subclass being called SBEII-2B. The remaining sequences are all of a completely new class of wheat SBEII genes, referred to herein as SBEII-1. The sequences have been found to fall into 3 sub-classes, to be discussed below. 
     The novel wheat SBEII-1 genes that are the subject of this invention have strong sequence homology with the maize SBEIIb gene. The wheat SBEII-1 genes are thought to have similar functional properties to the maize SBEIIb gene. On this basis it is expected that by genetic manipulation of the wheat SBEII-1 gene it will be possible to influence properties of starch produced by a plant, including the gelatinisation temperature and rheological properties of starch, in a manner analogous to manipulation of the maize SBEIIb gene described in WO 97/22703. The content of WO 97/22703 is incorporated herein by reference. 
     The present invention also includes within its scope a portion of any of the above sequences, comprising at least 500 base pairs and having at least 90% sequence homology to the corresponding portion of the sequence from which it is derived. 
     Although the coding sequences of the novel wheat SBEII-1 genes have strong sequence homology with the maize SBEIIb gene, there is much greater divergence in the 3′ untranslated parts of the sequences, with a maximum of 31.8% homology between the 3′ untranslated sequences of wheat SBEII-1 and maize SBEIIb as is apparent from FIG.  8 . 
     In another aspect the invention thus provides a nucleotide sequence comprising substantially the sequence shown in FIG. 5 (SEQ ID No: 8), FIG. 6 (SEQ ID No: 9) or FIG. 7 (SEQ ID No: 10), or a functional equivalent thereof. 
     The term functional equivalent in this context has the same general meaning as discussed above, but with equivalents preferably at least 32%, more preferably at least 40%, 50%, 60%, 70%, 80% or 90% sequence homology with the sequence of the relevant Figure. 
     It is thought such 3′ untranslated sequences may be useful, both in sense and antisense function, in manipulation of starch properties by affecting SBE expression in plants, as will be discussed below. 
     The sequence may include further nucleotides at the 5′ or 3′ end. For example, for ease of expression, the sequence desirably also comprises an in-frarne ATG start code, and may also encode a leader sequence. 
     The invention also covers a nucleic acid construct comprising a nucleotide sequence or portion thereof in accordance with the invention conveniently operably linked, in sense or antisense orientation, to a promoter sequence. 
     Also included within the scope of the invention is amino acid sequence encoded by any of the nucleotide sequences of the invention. 
     The invention also provides vectors, particularly expression vectors, comprising the nucleotide sequence of the invention. The vector will typically comprise a promoter and one or more regulatory signals of the type well known to those skilled in the art. The invention also includes provision of cells transformed (which term encompasses transduction and transfection) with a vector comprising the nucleotide sequence of the invention. 
     Nucleotide sequences in accordance with the invention may be introduced into plants, particularly but not exclusively wheat plants, and it is expected that this can be used to affect expression of SBE in the plant and hence affect the properties of starch produced by the plant. In particular, use of sequences in antisense orientation is expected to reduce or suppress enzyme expression. Additionally, it has recently been demonstrated in other experimental systems that “sense suppression” can also occur (i.e. expression of an introduced sequence operably linked in the sense orientation can interfere, by some unknown mechanism, with the expression of the native gene), as described by Matzke &amp; Matzke 1995. Any one of the methods mentioned by Matzke &amp; Matzke could, in theory, be used to affect the expression in a host of a homologous SBE gene. 
     It is believed that antisense methods are mainly operable by the production of antisense mRNA which hybridises to the sense mRNA, preventing its translation into functional polypeptide, possibly by causing the hybrid RNA to be degraded (e.g. Sheehy et al., 1988; Van der Krol et al.,). Sense suppression also requires homology between the introduced sequence and the target gene, but the exact mechanism is unclear. It is apparent however that, in relation to both antisense and sense suppression, neither a full length nucleotide sequence, nor a “native” sequence is essential. Preferably the “effective portion” used in the method will comprise at least one third of the full length sequence, but by simply trial and error other fragments (smaller or larger) may be found which are functional in altering the characteristics of the plant. 
     Thus, in a further aspect the invention provides a method of altering the characteristics of a plant, comprising introducing into the plant an effective portion of the sequence of the invention operably linked to a suitable promoter active in the plant so as to affect expression of a gene present in the plant. Conveniently the sequence will be linked in the antisense orientation to the promoter. Preferably the plant is a wheat plant. Conveniently, the characteristic altered relates to the starch content and/or starch composition of the plant (i.e. amount and/or type of starch present in the plant). Preferably the method of altering the characteristics of the plant will also comprise the introduction of one or more further sequences, in addition to an effective portion of the sequence of the invention. The introduced sequence of the invention and the one or more further sequences (which may be sense or antisense sequences) may be operably linked to a single promoter (which would ensure both sequences were transcribed at essentially the same time), or may be operably linked to separate promoters (which may be necessary for optimal expression). Where separate promoters are employed they may be identical to each other or different. Suitable promoters are well known to those skilled in the an and include both constitutive and inducible types. Examples include the CaMV 35S promoter (e.g. single or tandem repeat) and the ubiquitin promoter. Advantageously the promoter will be tissue-specific. Desirably the promoter will cause expression of the operably linked sequence at substantial levels only in the tissue of the plant where starch synthesis and/or starch storage mainly occurs. 
     The sequence of the invention, and the one or more further sequences if desired, can be introduced into the plant by any one of a number of well-known techniques (e.g. Agrobacterium-mediated transformation, or by “biolistic” methods). The sequences are likely to be most effective in affecting SBE activity in wheat plants, but theoretically could be introduced into any plant. Desirable examples include pea, tomato, maize, rice, barley, sweet potato and cassava plants. Preferably the plant will comprise a natural gene encoding an SBE molecule which exhibits reasonable homology with the introduced nucleic acid sequence of the invention. 
     In another aspect, the invention provides a plant cell, or a plant or the progeny thereof, which has been altered by the method defined above. The progeny of the altered plant may be obtained, for example, by vegetative propagation, or by crossing the altered plant and reserving the seed so obtained. The invention also covers parts of the altered plant, such as storage organs. Conveniently, for example, the invention covers grain comprising altered starch, said grain being obtained from an altered plant or the progeny thereof. Grain obtained from altered plants (or the progeny thereof) will be particularly useful materials in certain industrial applications and for the preparation and/or processing of foodstuffs and may be used, for example, in bakery products. 
     In particular relation to wheat plants, the invention provides a wheat plant or part thereof which, in its wild type possesses an effective SBEII-1 gene, but which plant has been altered such that there is either reduced, increased or no effective expression of an SBEII-1 polypeptide within the cells of at least part of the plant. The plant may have been altered by the method defined above, or may have been selected by conventional breeding to be deleted for the SBEII-1 gene, the presence or absence of which can be readily determined by screening samples of the plants with a nucleic acid probe or antibody specific for the wheat gene or gene product respectively. 
     The invention also provides starch extracted from a plant altered by the method defined above, or from the progeny of such a plant, the starch having altered properties compared to starch extracted from equivalent, but unaltered, plants. The invention further provides a method of making altered starch, comprising altering a plant by the method defined above and extracting therefrom starch having altered properties compared to starch extracted from equivalent, but unaltered, plants. It is believed that use of nucleotide sequences in accordance with the invention will enable the production of starches, particularly wheat starches, having a wide variety of novel properties. For example, it may be anticipated that plants altered to give a reduction in SBEII activity will give rise to a starch with a relatively higher proportion of amylose and a lower proportion of amylopectin compared with that from unaltered plants. 
     In particular the invention provides the following: a plant (especially a wheat plant) altered by the method defined above, containing starch which, when extracted from the plant, has an elevated gelatinisation onset and/or peak temperature as measured by DSC, compared to starch extracted from a similar, but unaltered, plant; a plant (especially a wheat plant) altered by the method defined above, containing starch which, when extracted from the plant, has a elevated gelatinisation onset temperature (conveniently elevated by at least 3° C., possibly by at least 7° C., by at least 12° C. or possibly even by 15 to 25° C.) as measured by DSC compared to starch extracted from a similar, but unaltered plant; a plant (especially a wheat plant) altered by the method defined above, particularly to reduce expression of SBEII-1 polypeptide, containing starch which, when extracted from a plant, has a higher amylose:amylopectin ratio compared to starch extracted from a similar, but unaltered plant. 
     The present invention particularly covers starch extracted from a plant altered by the method of the invention, particularly starch having an increased gelatinisation temperature. Such starch is useful, eg in bakery products, having particular benefits in certain situations, and the invention also covers products, particularly bakery products, made from such starch. The invention also covers starch extracted from a plant altered by the method of the invention and having an increased amylose:amylopectin ratio. 
     The invention will be further described, by way of illustration, in the following Examples and with reference to the accompanying drawings, in which: 
    
    
     BRIEF DESCRIPTION OF THE DRAWINGS 
     FIG. 1 is a graph of viscosity versus time, showing a viscoamylgraph profile for wheat starch during and after cooking; 
     FIGS  2 ( i )- 2 ( vi ) show alignment amino acid sequence data of C terminal portions of various known starch branching enzymes (SEQ ID Nos: 12 to 25), obtained from the European Molecular Biology Laboratory (EMBL) database, and for a novel wheat SBEII-1 sequence of the invention (OsbeII-1ALL) (SEQ ID No: 11) from clone 5A1, with consensus residues highlighted; 
     FIG. 2 a  is a residue weight table showing the percent similarity and percent divergence of the sequences shown in FIG. 2; 
     FIGS.  3 ( i )- 3 ( vi ) show aligned DNA sequence data for various recombinant clones (B2, B4, B10, A2, B1, B11) (SEQ ID Nos: 3, 4, 5, 26, 6, 27 respectively) containing wheat starch branching enzyme genes, representing two SBE classes, SBEII-1 and SBEII-2, each of which includes three subclasses A, B and C, with residues differing from the consensus (majority) (SEQ ID No: 53) highlighted; 
     FIG. 3 a  is a residue weight table showing the percent similarity and percent divergence of the sequences shown in FIG. 3; 
     FIG. 4 is an alignment of predicted amino acid sequences for clones B6 (wheat SBEII-1) (SEQ ID No: 7) and B11 (wheat SBEII-2) (SEQ ID No: 28) against the corresponding regions of the maize SBEIIa (SEQ ID No: 29) and SBEIIb (SEQ ID No: 30) amino acid sequences, with residues differing from those of maize SBEIIb highlighted; 
     FIG. 4 a  is a residue weight table showing the percent similarity and percent divergence of the sequences shown in FIG. 4; 
     FIG. 5 shows the 3′ untranslated DNA sequence of clone B2 (SEQ ID No: 8) (wheat SBEII-1, sub-class A); 
     FIG. 6 shows the 3′ untranslated DNA sequence of clone B10 (SEQ ID No: 9) (wheat SBEII-1, sub-class B); 
     FIG. 7 shows the 3′ untranslated DNA sequence of clone B4 (SEQ ID No: 10) (wheat SBEII-1, sub-class C);. 
     FIGS.  8 ( i )- 8 ( iii ) show aligned DNA sequence data for the 3′ untranslated region of clones B10 (SEQ ID No: 9), B2 (SEQ ID No: 8) and B4 (SEQ ID No: 10) and maize SBEIIb (ZMSBE2b) (SEQ ID No: 31), with residues differing from those of the B10 sequence highlighted; 
     FIG. 8 a  is a residue weight table showing the percent similarity and percent divergence of the sequences shown in FIG. 8; 
     FIGS. 9 a  and  9   b  show hybridisation of clone B1 (SBEII-2) and clone B2 (SBEII-1), respectively, to HindIII-digested genomic DNA of Chinese Spring wheat nullisomic-tetrasomic lines; 
     FIGS.  10 ( i )- 10 ( iii ) show the DNA (SEQ ID No: 1) and predicted amino acid sequence (SEQ ID No: 2) of part of SBEII-1 clone 5A1; 
     FIGS.  11 ( i )- 11 ( v ) show aligned amino acid sequence data for the wheat SBEII-1 sequence of the invention, from clone 5AI (OsbeII-1ALL) (SEQ ID No: 11), wheat SBEI-D2 (SEQ ID No: 32) of Rahman et al 1997 (TASBEID2), wheat SBE1 of Rapellin et al 1997 (SEQ ID No: 33) (TASBEI) and wheat SBEII-2 of Nair et al 1997 (SEQ ID No: 34) (wheat SBEII-2), with residues exactly matching the consensus (majority) (SEQ ID No: 54) highlighted; 
     FIG. 11 a  is a residue weight table showing the percent similarity and percent divergence of the sequences shown in FIG. 11; 
     FIG. 12 illustrates northern blotting of wheat grains harvested at various different intervals after anthesis and probed with SBEII-1 and SBEII-2 fragments; 
     FIG. 13 is a restriction map of plasmid pWxGS+; 
     FIG. 13 a  shows the sequence (SEQ ID No: 55) of the promoter (HindIII-BamH1 fragment) in pWxGS+; 
     FIG. 14 is a restriction map of plasmid pSRWXGUS1; 
     FIG. 15 is a restriction map of plasmid pVTWXGUS2; 
     FIG. 16 is a restriction map of plasmid pPBI-97-2; 
     FIG. 17 is a restriction map of plasmid pSR97-26A−; 
     FIG. 18 is a restriction map of plasmid pSR97-29A−; 
     FIG. 19 is a restriction map of plasmid pSR97-50A−; 
     FIG. 20 is a restriction map of plasmid pSR97-53A−; 
     FIG. 21 is a restriction map of plasmid p97-2C; 
     FIG. 22 is a restriction map of plasmid p97-2CWT1; 
     FIG. 23 is a restriction map of plasmid pSC98-1; 
     FIG. 24 is a restriction map of plasmid pSC98-2; 
     FIG. 25 is a restriction map of plasmid pUNI; 
     FIG. 26 shows the DNA sequence of the NptII Sac1 fragment of pUNI (SEQ ID No: 35); and 
     FIG. 27 is a restriction map of plasmid pUSN99-1; 
     FIG. 28 is a restriction map of plasmid pUSN99-2; 
     FIGS.  29 ( i )- 29 ( iv ) show is a partial restriction map of the predicted sequence (SEQ ID No: 52) of a cloned fragment of p97-U3; 
     FIG. 30 is a restriction map of plasmid pPBI96-36; 
     FIG. 31 is a restriction map of plasmid p97-dUG1; 
     FIG. 32 is a restriction map of plasmid p97-2BdUN1; 
     FIG. 33 is a schematic illustration of a particle bombardment chamber (not to scale); 
     FIG. 34 shows histochemical localisation of Ubi-GUS expression in seed (panel A), stem (panel B), floral (panel C) and leaf tissues (panel D) of wheat transformed with plasmid pAHC25; 
     FIG. 35 is a Southern blot of 26 progeny plants of transformant BW119 which had been transformed with pAHC25. 
     FIG. 36 shows histochemical localisation of waxy-GUS expression in endosperm tissue of two independent transgenic wheat lines (in panels A and B) transformed with the plasmid pWxGS+; and 
     FIG. 37 is a Southern blot of genomic DNA of putative primary transformants digested with Sac1 and probed with the 1 kb Sac1 SBEII-1 probe. 
    
    
     DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS 
     EXAMPLES 
     Amplification and Characterisation of Two Classes of SBEII cDNA Clones 
     A PCR based cloning strategy was devised for isolating starch branching enzymes from wheat using conserved domains within the known cloned gene sequences. Starch branching enzymes have been cloned from a number of plant species and FIG. 2 shows amino acid sequence data, obtained from the European Molecular Biology Laboratory (EMBL) nucleotide database for various known starch branching enzymes as follows: 
     Wheat SBEII-2 for  Triticum aestivum  (SEQ ID No: 12) 
     ZM SBE2a (maize) for  Zea mays  (SEQ ID No: 13) 
     ZM SBE2b (maize) for  Zea mays  (SEQ ID No: 14) 
     Barley SBEIIa (SEQ ID No: 15) 
     Barley SBEIIb (SEQ ID No: 16) 
     RICBCE3 (rice SBEII type enzyme) for  Oryza saliva  (SEQ ID No: 17) 
     RICESBE-1/97 (as above, including transit peptide sequence) (SEQ ID No: 18) 
     PSSBEIGEN (pea SBEI, which is in fact an SBEII- type sequence) for  Pisum sativum  (SEQ ID No: 19) 
     STSBE (potato SBEI type) for  Solanum tuberosum  (SEQ ID No: 20) 
     TASBEI (wheat SBEI-2) for  Triticum aestivum  (SEQ ID No: 21) 
     TASBEI D2 (SEQ ID No: 22) 
     ZMSBEI (maize SBEI) for i Zea mays (SEQ ID No: 23) 
     RICBEI (rice SBEI) for  Oryza sativa  (SEQ ID No: 24) 
     PSSBEIIGN (pea SBEII, which is in fact an SBEI-type sequence) for  Pisum sativum  (SEQ ID No: 25) 
     FIG. 2 also shows sequence information for a novel wheat SBEII-1 sequence of the invention, identified as OsbeII-1ALL (SEQ ID No: 11). 
     The alignment report of FIG. 2, and also FIGS. 3,  4 ,  8  and  11 , was prepared using Clustal method, with PAM 250 residue weight table for amino acid sequences and weighted residue weight table for DNA sequences. Sequence pair distances expressed as % similarity shown in FIGS. 2A and 3A,  4 A,  8 A and  11 A are determined using a ‘MEGALIGN’ program of DNAStar software, and correspond to sequence homology percentages as specified above. 
     Alignment of the sequences shown in FIG. 2 reveals several domains which are highly conserved. One such domain, MDKDMYD (SEQ ID No: 36), was almost completely conserved and it was assumed that this domain would also be present in wheat starch branching enzyme genes. This motif was chosen as a target for an oligonucleotide sense primer (SBEA). 3′RACE PCR was carried out on endosperm first strand cDNA using the primers Ro and SBE A. 
     Two populations of PCR products of approximately 1 kb and 1.2 Kb were cloned into the plasmid vector pT7Blue (Novagen). Plasmid DNA from 36 putative recombinant clones was purified and the insert size estimated by restriction analysis. Fifteen clones harbouring inserts of between approximately 1 Kb and 1.2 Kb were selected for sequencing. Alignment of the sequence data obtained, using the MEGALIGN program of DNAStar, indicated that the 15 selected clones could be divided on the basis of degrees of homology into two different classes, which we have designated SBEII-1 and SBEII-2. Furthermore, both the SBEI1-I and SBEII-2 classes may each be further subdivided into three sub-classes, based on sequence differences (Table 1). It is thought the sub-division into three sub-classes probably arises because wheat comprises three homoeologous genomes. 
     
       
         
               
               
               
               
             
               
               
               
               
             
           
               
                   
                 TABLE 1 
               
               
                   
                   
               
               
                   
                 Class 
                 Sub-Class 
                 Clone Number 
               
               
                   
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 SBEII-1 
                 A 
                 B2, B5, B6, B7, B12 
               
               
                   
                 SBEII-1 
                 B 
                 B10 
               
               
                   
                 SBEII-1 
                 C 
                 A1, A13, B4 
               
               
                   
                 SBEII-2 
                 A 
                 B11 
               
               
                   
                 SBEII-2 
                 B 
                 B1, B9 
               
               
                   
                 SBEII-2 
                 C 
                 A2, C5 
               
               
                   
                   
               
             
          
         
       
     
     Comparison between sequences within either of the SBEII-1 or SBEII-2 classes showed between 90 and 96.8% similarity. In contrast, sequence similarity between representatives of SBEII-1 and SBEII-2 classes only display between 58.8 and 60.0% homology in the region of comparison (FIGS. 3 and 3 a ). 
     Furthermore, we have compared representative sequences from each SBEII-1 and SBEII-2 class with the previously reported wheat SBEII clones, pWBE6 (Mousley, 1994) and the very recently published SBEII (Nair et al., 1997). The results showed that each of the previously isolated SBEII clones are highly homologous (&gt;90%) to our SBEII-2 class (data not shown). Significantly, neither of the previously reported wheat sequences showed high homology to our SBEII-1 sequence. The isolation and characterisation of three forms of SBEII-1 (SBEII-1, sub-classes A, B &amp; C) is novel. The SBEII-2 sub-class B is also novel, sub-classes A and C corresponding to the sequences previously disclosed by Mousley (1994) and Nair et al (1997) respectively. 
     Alignment of the predicted amino acid sequences from representative clones, B6 and B11 of the wheat SBEII-1 and SBEII-2 sequences (respectively) against the corresponding regions of the maize SBEIIa and SBEIIb amino acid sequences (FIGS. 4 and 4 a ) indicate that the wheat SBEII-1 sequence (clone B6) is more similar to the maize SBEIIb sequence (88.7% similarity) than to the wheat SBEII-2 sequence and the maize SBEIIa sequence (82.2% &amp; 82.6% similarity respectively) and similarly that the wheat SBEII-2 sequence is more similar to the maize SBEIIa sequence (86.9% similarity) than to the wheat SBEII-1 and maize SBEIIb sequences (82.2% and 81.7% similarity respectively). We thus hypothesise that the wheat SBEII-1 is phylogenetically more related to the maize SBEIIb and that the wheat SBEII-2 is phylogenetically related to the maize SBEIIa sequences and that the corresponding wheat and maize sequences are likely to exhibit similar functional properties. 
     While the coding sequences of clones B2, B10 and B4 have strong sequence homology to the maize SBEIIb gene, there is much greater divergence in the 3′ untranslated parts of the sequences. FIGS. 5,  6  and  7  show the 3′ untranslated sequences of clones B2, B10 and B4, respectively, and FIG. 8 compares these sequences with the corresponding sequence of maize SBEIIb. 
     Considering matters in more detail, experimental details were as follows. 
     Plant Material 
       Triticum aestivum  cultivar Rialto was grown in a glass house under supplementary lighting and temperature control to maintain a 16 hour day-length at 18+/−1° C. 
     Recombinant DNA Manipulations and Sequencing 
     Standard procedures were performed essentially according to Sambrook et al., (1989). DNA sequencing was performed on an ABI automated sequencer and sequences analysed using DNASTAR software for Macintosh. 
     RNA Isolation for cDNA Cloning 
     RNA was extracted from  Triticum aestivum  cultivar Rialto endosperm, using a Purescript RNA isolation kit (Flowgen) essentially according to the manufacturers recommendations. Briefly, endosperm tissue was frozen in liquid nitrogen and ground, for 2 min, to a fine powder using a dismembrenator (Braun Biotech International). The ground tissue was stored in liquid nitrogen prior to extraction. Approx. 100 mg of ground tissue was transferred to a 1.5 ml microcentrifuge tube and 1.2 ml of ‘Lysis buffer’ was added to the tissue before mixing by inversion and placing on ice for 10 minutes. Protein and DNA were precipitated from the cell lysate by adding 0.4 ml of ‘Protein-DNA Precipitation Solution’ and mixing by inversion before centrifuging at 13,000×g at 4° C. for 20 minutes. The supernatant was divided between two fresh 1.5 ml tubes each containing 600 μl of isopropanol. The RNA precipitate was pelleted by centrifugation at 13,000×g at 4° C. for 10 minutes, the supernatant was discarded and the pellets washed with 70% ethanol by inverting the tube several times. The ethanol was discarded and the pellet air dried for 15-20 minutes before the RNA was resuspended in 7.5 ml of ‘RNA Hydration Solution’. 
     Preparation of Wheat Endosperm cDNA Pool 
     Wheat endosperm cDNA pool was prepared from total RNA, extracted as described above, using Superscript™ reverse transcriptase (Life Technologies) essentially according to manufacturers instructions. Briefly, five microgrammes of RNA. 10 pMol RoRidT17 [AAGGATCCGTCGACATCGATAATACGACTCACTATAGGGA(T17)] (SEQ ID No: 37) and sterile distilled water to a reaction volume of 12 μl, in a 500 μl microcentrifuge tube, was heated to 70° C. for 10 minutes before being quick chilled on ice. The contents of the tube were collected by brief centrifugation before adding 4 μl 5× First Strand Buffer, 2 μl 0.1M DTT and 1 μl 10 mM dNTPs and, after mixing, incubating at 42° C. for 2 min. 1 μl of Superscript™ was added and, after mixing, incubation continued for 1 hour. The reaction was inactivated by heating to 70° C. for 15 min. 150 μl of T 10 E 1  was added to the reaction mix and the resulting cDNA pool was used as a template for amplification in PCR. 
     PCR Amplification of SBEII Sequences from Endosperm cDNA Pool 
     SBEII sequences were amplified from the endosperm cDNA pool using primers Ro [AAGGATCCGTCGACATC] (SEQ ID No: 38), which is complementary to the Ro region of the RoRidT17 primer used to synthesise the cDNA pool, and the SBEII specific primer, SBEA [ATGGACAAGGATATGTATGA] (SEQ ID No: 39). SBEA was designed to be homologous to the MDKDMYD (SEQ ID No: 36) motif which is situated approx. 1 kb from the 3′ end of the mature peptide coding sequence. PCR was carried out in a 50 μl reaction, comprising 5 μl of the cDNA pool, 25 pmol Ro, 50 pmol SBEA, 5 μl 5× Taq buffer, 4 μl 25 mM Mg 2+ , 0.5 μl 20 mM dNTPs, and 1.25 u Taq polymerase. All of the reaction components were mixed, except for the Taq polymerase, before being pre-heated to 94° C. for 7 min and then cooled to 75° C. for 5 min. Whilst the reaction mixtures were held at 75° C. the Taq polymerase was added and, after mixing well, the reactions were thermocycled at (94° C.—30 sec, 50° C.—30 sec, 72° C.—1 min)×30 cycles, followed by a final 10 min extension step at 72° C. 
     PCR products were purified by phenol/chloroform and chloroform extraction before ligation with pT7 Blue (Novagen) according to manufacturers recommendations. Putative SBE clones were initially characterised by standard plasmid DNA purification methods and restriction digestion. Representative clones harbouring a range of different sized inserts were selected for sequencing. 
     Chromosomal Location of SBE Genes in Wheat 
     The Chinese Spring wheat nullisomic-tetrasomic lines as described in Sears (1966) were used for assignment of the SBE sequences chromosome locations. Ditelosomic lines (Sears, 1966) were used to determine the chromosome arm location. The Betzes barley ditelosomic addition lines in wheat are described in Islarn (1983). 
     The chromosomal location of the two families of SBEII sequences (SBEII-1, SBEII-2) was determined by probing wheat nulli-tetra and ditelosomic stock lines with gel-purified inserts of the various clones. FIG. 9 a  shows the hybridisation obtained with an SBEII-2 (clone B1) probe on HindIII digested DNA. The euploid Chinese Spring gives 3 bands, one of which is missing in turn in the lines nullisomic for chromosomes 2A, 2B and 2D. The same blot was re-probed with a SBEII-1 specific probe (clone B2). This yields an entirely different hybridisation profile (FIG. 9 b ), demonstrating the specificity of the probe used. Again bands are missing in each of the lines nullisomic for 2A, 2B and 2D the same banding pattern was observed using the SBEII-1 clones B2 and B4. Thus the SBEII sub-family 1 and 2 gene sequences lie on the wheat group 2 set of homeologous chromosomes. 
     Ditelosomic addition lines were used to identify the arm location of these genes (data not shown). This revealed that the SBEII-1 and SBEII-2 sequences are both located on the long arms of the homeologous group 2 chromosomes of wheat. 
     Barley addition lines were used to determine whether homologous sequences are present in barley. These showed that sequences homologous to the wheat SBEII-1 and SBEII-2 sequences are located on the long arms of barley chromosome 2H. 
     RNA Isolation and Northern Blotting 
     Wheat grains were harvested at appropriate intervals and frozen in liquid Nitrogen before grinding to a fine powder using either a Braun Mikrodismembrator™ or a pestle and mortar. Total RNA was isolated using the RNAqueous™ (Ambion Inc) Kit according to the manufacturers instructions, or with the following method. Frozen powdered grain was mixed with a 10× volume of 0.2M Tris-HCl pH9, 0.4M NaCl, 25 mM EDTA, 1% SDS, 1% PVPP, 0.25% Antifoam A, and 0.1M DTT. This mixture was extracted twice with an equal volume of phenol/chloroform/isoamyl alcohol (25:24:1), the nucleic acids precipitated from the aqueous phase by the addition of 0.8 volumes of isopropanol, and the resulting pellet dissolved in H 2 O. The RNA was then selectively precipitated by the addition of 1 volume of 4M LiCl, incubated at 4° C. overnight, and the resulting pellet dissolved in sterile distilled H 2 O. 15 μg of total RNA was electrophoresed on a 1% agarose, 2.21M Formaldehyde, 40 mM MOPS pH7.0, 10 mM sodium acetate, 1 mM EDTA gel, in a 40 mM MOPS pH7, 10 mM sodium acetate, 1 mM EDTA running buffer at 1 V/cm overnight. Gels were placed in a 50 ng/ml solution of Ethidium Bromide in water for 30 minutes, de-stained in water for 2 hours, and visualised and photographs under UV light. The gels were then washed briefly in sterile distilled H 2 O, then blotted onto HyBond N + ™ (Amersham International), according to standard protocols (Sambrook et al, 1989) overnight. Blots were then dismantled and air-dried before UV fixing at 312 nm for 2 minutes. 
     Probe Isolation and Purification 
     5-10 μg of the plasmids pUN1 and pSR98-29 were digested with Sst1 (Life Technologies Ltd) according to the manufacturers instructions, to release fragments of approximately 0.8kb (NptII) and 1 kb (SBEII-1) respectively. 5-10 μg of the plasmid pVT96-54 was digested with BamH1 to release a SBEII-2 fragment of approximately 1.2 kb. Digests were electrophoresed on 1% low melting point agarose gels. The gene specific fragments were excised and the DNA purified using a Wizard™ Gel Purification Kit (Promega). 
     Probe Labelling and Hybridization 
     25 ng of the appropriate probe (Maize Waxy promoter, NptII, Wheat SBEII-1 or Wheat SBEII-2 fragments) were radiolabelled using the Rediprime 11™ system (Amersham International) using α 32 PdCTP (Amersham International) according to manufacturers instructions. Blots were hybridized overnight at 65° C. in 0.6M NaCl, 20 mM Pipes, 4 mM Na 2 EDTA2H 2 O, 0.2% gelatin, 0.2% Ficoll 400, 0.2% PVP-360, 10 mM Na 4 P 2 O 7 10H 2 O, 0.8% SDS, 0.5 mg/ml denatured salmon sperm DNA. Post hybridization washes were carried out in 30 mM NaCl, 2 Mm NaH 2 PO 4 .2H 2 O, 0.2 mM Na 2 EDTA.2H 2 O, 0.1% SDS at room temperature for 7 minutes, then 65° C. for 10 minutes. Filters were exposed to Kodak BioMax MR™ (Amersham International) film at −70° C. Blots were stripped by washing in 15 mM NaCl, 1 mM NaH 2 PO 4 .2H 2 O, 0.1 mM EDTA at 90° C. for 10 minutes, or until no counts above background remained. 
     Extension of the SBEII-1 3′ Sequence Towards the 5′ End of the Mature Peptide 
     We have exploited the sequence divergence between our wheat SBEII-1 and SBEII-2 sequences to design the SBEII-1 specific 3′ primer, Sb4. This primer was used in conjunction with an SBEII specific 5′ primer to extend the novel SBEII-1 sequence using a PCR-based approach. 
     To extend the SBEII-1 3′ sequence towards the 5′ end of the mature peptide, a second conserved domain was identified and an oligonucleotide sense primer, AGSBEI, designed. PCR amplification from the endosperm first strand cDNA pool was carried out using the AGSBEI-Sb4 primer pair. Separation of the amplification products by electrophoresis through a 1% (w/v) agarose gel (data not shown) showed that the reaction yielded a distinct band of approx. 2.2 kb. The approx 2.2 kb amplification products were excised from the gel, ligated with PT7Blue and transformed into competent Novablue  E. coli  cells. Following overnight culture, nine putative recombinant clones were selected for further analysis. Screening of each of the selected clones using vector specific primers indicated that clones 5A1, 5A2, 5A5 and 5A9 harboured inserts of the predicted size. Of these clone 5A1 (which falls in sub-class C) was selected for sequencing (FIG.  10 ). The amino acid sequence of FIG. 10 corresponds to the OsbeII-1ALL sequence of FIG.  2 . Although not full length the predicted open reading frame includes nucleotides 44 through to 1823 and encodes a 593 amino acid peptide. Based on similarities with the maize genes, it is estimated that this sequence is missing approximately 230 amino acids out of a predicted total of approximately 830 amino acids. On this basis, the partial sequence represents about 70% of the coding sequence. Multiple sequence alignment of this SBEII-1 sequence with recently published wheat SBEII-2 (Nair et al., 1997), SBEI (Rapellin et al., 1997) and SBEI-D2 (Rahman et al., 1997) sequences showed that the SBEII-1 sequence has similarity indices of 69.6%, 31.2% and 46.7% to SBEII-2, SBEI and SBEI-D2 respectively (FIGS. 11 and 11 a ). This demonstrates that the SBEII-1 sequence differs from the published wheat SBE sequences, and confirms the analysis of the 3′ sequence alignment (FIG.  3 ). The increase in relative homology when compared to the values obtained following 3′ sequence alignment results from the fact that the central domain of SBEs is highly conserved (Burton et al., 1995; Gao et al., 1997). However, it is clear that this cloned wheat SBEII-1 sequence is significantly different from previously published wheat SBE sequences and represents a novel sequence. 
     Full experimental details were as follows. 
     SBEII-1 sequences were extended toward the 5′ end of the mature peptide by amplification from the endosperm cDNA pool using the SBEII-1 specific primer Sb4 [TTTTCTCACAACGCCCTGGG] (SEQ ID No: 40) in conjunction with the primer AGSBEI [TGTTTGGGAGATCTTCCTCCC] (SEQ ID No: 41). AGSBEI was designed to be homologous to the GVWEIFLP (SEQ ID No: 42) motif which is conserved in all known SBE sequences and is situated toward the 5′ end of the mature peptide coding sequence. PCR was carried out in a 50 μl reaction, comprising 5 μl of the cDNA pool, 50 pmol Sb4, 50 pmol SBEA1, 5 μl 5× Taq buffer, 4 μl 25 mM Mg 2+ , 0.5 μl 20 mM dNTPs, and 1.25 u Taq polymerase. All of the reaction components were mixed, before thermocycling at (94° C.—45 sec, 55° C.—30 sec, 72° C.—1 min 30 sec)×30 cycles, followed a final 10 min extension step at 72° C. Amplification products were separated by electrophoresis through a 1%(w/v) agarose gel and specific amplification products of the predicted size were excised from the gel. The DNA was eluted from the gel slice using QIAGEN&#39;s gel extraction kit according to the manufacturers recommendations before ligation with pT7 Blue (Novagen). Ligation was carried out in a 10 μl reaction volume comprising 7.5 μl purified amplification product, 1 μl 10× ligation buffer, 1 μl pT7Blue and 0.5 μl T4 DNA ligase (Amersham). The reaction components were mixed well before being placed at 4° C. overnight. Following overnight incubation, half of the ligation reaction was used to transform competent Novablue  E.coli  cells (Novagen). Transformed cells were plated out onto LB plates supplemented with X−gal (40 μgml −1 ), IPTG (0.1 mM), Carbenicillin (100 μgml −1 ), and Tetracycline (12 μgml −1 ), before placing at 37° C. overnight. Putative recombinant clones were initially screened for the presence of an insert by colony PCR using the vector specific primers T7B and U19. Insert positive clones were then screened using an insert specific primer in conjunction with either T7B or U19 primers to determine the orientation of the insert within the multiple cloning site prior to sequencing. 
     Southern Blot Analysis 
     Southern analyses of the pre-made nulli-tetra and ditelosomic blots were carried out essentially as described in Jack et al (1994). 
     The SBEII-1 clones discussed above have been cloned into transformation vectors for transformation of wheat. 
     Northern Blot Analysis 
     Northern blots were prepared from total RNA from developing wheat grains of the cultivar Bobwhite. FIG. 12 shows a northern blot of RNA from wheat grains of the cultivar Bobwhite grown in the glasshouse as described and harvested between 5 and 29 days after anthesis. The blot was probed with the 1 kb Sac1 SBEII-1 fragment and subsequently (following blot stripping) with the 1.2 kb BamH1 SBEII-2 fragment, both fragments purified and labelled as described. In FIG. 12 panel A shows the Ethidium Bromide-stained RNA gel prior to northern transfer. Panel B shows the results of probing with the SBEII-1 probe and panel C shows the results of probing with the SBEII-2 probe. Comparing within and between panels B and C differences can be observed in the relative intensities of the signals at the different time points. In particular a relatively stronger signal intensity is observed with the SBEII-2 probe for the 5 day time point than with the SBEII-1 probe, indicating that the transcript profiles for SBEII-1 and SBEII-2 are distinct, suggesting that the two gene families (SBEII-1 and SBEII-2) are differentially expressed during grain development. The size of the transcripts observed for both SBEII-1 and SBEII-2 is approximately 3.5 kb. However the SBEII-2 transcript is slightly smaller than the SBEII-1 transcript. 
     Plasmid Constructions 
     Standard molecular biology procedures (Sambrook et al, 1989) were used for plasmid constructions. 
     pWxGS+ (FIG. 13) comprising a maize granule bound starch synthase gene (Shure et al 1983) promoter-GUS-Nos fusion was obtained as a gift to Unilever Research from Sue Wessler (University of Georgia, Athens, USA) and may be obtained on request from that source. The promoter in pWxGS+ is approximately 1.5 kb in length and represents a truncated version of a similar, but larger promoter fragment described in Russell &amp; Fromm (1997). The sequence of the promoter (HindIII-BamH1 fragment) in pWxGS+ is presented in FIG. 13A (SEQ ID No: 55). 
     pSRWXGUS1 (FIG. 14) was produced by inserting a Sac 1 linker [d(pCGAGCTCG)0] (New England Biolabs [NEB]) (NEB catalogue No 1044) into the Sma1 site in pWxGS+. 
     pVTWXGUS2 (FIG. 15) was produced by inserting a BamH1 linker [d(pCGGGATCCCG)] (SEQ ID No: 43) (NEB catalogue No. 1071) into the Ec1136II (an isoschizomer of Sac1 which gives blunt ends) site of pWxGS+. 
     A Sac1 linker was inserted at the XbaI site (which had been blunted using Klenow+dNTps) of the SBEII-1 Clone B6 in the plasmid pT7Blue to produce an intermediate clone. The SBE sequence was then purified from this intermediate clone as a Sac1 fragment and ligated into the Sac1 sites of pSRWXGUS1 replacing the GUS gene sequence to produce the plasmids pSR96-26 and pSR96-29 representing antisense and sense orientations of the SBEII-1 sequence downstream of the Waxy promoter, respectively. 
     A BamH1 linker was inserted at the XbaI site (which had been blunted using Klenow+dNTps) of the SBEII-2 Clone B11 in pT7Blue to produce an intermediate clone. The SBE sequence was then purified from this intermediate as a BamH1 fragment and inserted into the BamH1 sites of pVTWXGUS2, replacing the GUS gene sequence, to produce the plasmids pVT96-50 and pVT96-53 representing antisense and sense orientations, respectively, of the SBEII-2 sequence downstream of the Waxy promoter. 
     pVT96-54. A BamH1 linker was inserted at the Xba1 site (which had been blunted using Klenow+dNTPs) of the SBEII-2 clone B9 (equivalent to clone B1) in pT7Blue to produce an intermediate clone. The SBEII-2 sequence was then purified from this intermediate clone as a BamH 1 fragment and inserted into the BamH1 sites of pVTWXGUS2, replacing the GUS gene sequence, to produce the plasmid pVT96-54. 
     The Waxy-SBE-NOS sequences in the plasmids pSR96-26 and pSR96-29 and pVT96-50 and pVT96-53 were purified as HindIII/EcoRI fragments and inserted into the EcoRI/HindIII sites of plasmid pPBI-97-2 (also known as p97-2) (FIG.  16 ). Plasmid pPBI-97-2 is described in European Patent Application No. 97305694.8 (published as WO 99/06570). Following removal of the ampicillin resistance marker gene the resulting plasmids were designated pSR97-26A− (clone B6 (SBEII-1, sub-class A) in antisense orientation), pSR97-29A− (clone B6 in sense orientation), and pSR97-50A− (clone B11 (SBEII-2, sub-class A) in antisense orientation) and pSR97-53A− (clone B11 in sense orientation) as illustrated in FIGS. 17,  18 ,  19  and  20 , respectively. 
     p97-2C (FIG. 21) was produced by digesting the polylinker sites Ec1136 II to SmaI in the plasmid pPBI97-2 (FIG.  16 ), ligating and selecting recombinants in which the polylinker region from SmaI to Ec1136 II had reinserted in the opposite orientation. 
     The Waxy-NOS sequences in pSRWXGUS1 were transferred as a HindIII/EcoRI fragment into the HindIII/EcoRI sites of plasmid p97-2C to produce the plasmid p97-2CWT1 (FIG.  22 ). 
     pSC98-1 and pSC98-2. The 5′ extended SBEII-1 clone 5A1 in pT7Blue (comprising SBE sequence from coordinate 43 to 2003 bp in FIG. 10) was digested with EcoRI and Xba1, followed by ‘in-fill’ of overhangs using Klenow polymerase and dNTPs. The resulting blunt ended SBE fragment was gel purified and ligated to p97-2CWT1 (FIG. 22) which bad been digested with Ec1136II and dephophorylated using calf intestinal phosphatase. The resulting recombinants were screened by restriction digest anaylsis arid clones comprising both orientations of the SBE sequence (with respect to the waxy promoter) were identified. pSC98-1 (FIG. 23) is an antisense version and pSC98-2 (FIG. 24) is a sense version. Following removal of the ampicillin marker gene the resulting plasmids were designated pSC98-1A− and pSC98-2A− respectively. 
     Ubiquitin Promoter—NptII Selection Construct:pUN1 
     pUN1 was made in the following way: 
     A Sac1 linker was inserted at the SmaI site of the plasmid pAHC25 (Christensen and Quail 1996) to produce an intermediate plasmid. The GUS gene was removed from this intermediate plasmid by digesting with SacI followed by self ligation and identification of recombinant molecules lacking the GUS sequence to produce the plasmid pPBI95-9. pPBI95-9 was digested with EcoRI and following self ligation recombinant molecules lacking the Ubi-BAR sequences were identified. The resulting plasmid is designated pPBI96-23. An NptII sequence was amplified as a PCR product using the primers AG95-7: 5′GATGAGCTCCGTTTCGCATGATTGAACAAGATGG (SEQ ID No: 44) and AG95-8: 5′GTCGAGCTCAGAAGAACTCGTCAAGAAGGC (SEQ ID No: 45), using pPBIBAG3 (Goldsbrough et al 1994 as template for the NptII sequence. The amplified product was cloned into the SstI site of pBluescript (Stratagene) and sequenced. The sequencing revealed that the NptI sequence was of the ‘mutant’ form rather than the wild-type as had been expected. The ‘mutant’ form carries a single base change which is flanked by unique Nco1and Sph1 sites. The pBluescript clone was digested with Nco1 and Sph1 to remove the region containing the single base change. Two oligonucleotides, (Npt1:CCCGACGGCGAGGATCTCGTCGTGACC (SEQ ID No: 46) and Npt2: CATGGGTCACGACGAGATCCTCGCCGTCGGGCATG) (SEQ ID No: 47) were then annealed to each other to form an Nco1/Sph1 fragment. This was cloned into the Nco1/Sph1 digested Bluescript/NptII clone, and the resulting clone was sequenced to confirm that the gene was now of the wild type form. 
     The NptII sequences was then purified as a SacI fragment and inserted at the SacI site of pPBI96-23 to produce pUN1 (FIG.  25 ). pUN1 includes the wild-type ubiquitin promoter (Ubi promoter), which is also referred to as the ubiquitin regulatory system (abbreviated to URS). The orientation of the NptII sequence in pUN1 was determined by restriction digest analysis. The sequence of the NptII SacI fragment is presented in FIG. 26 (SEQ ID No: 35). 
     pUSN99-1 and pUSN99-2. The SBEII-1 (clone B6) sequence was purified as a Sac1 fragment from the plasmid pSR96-26 and inserted at the Sac1 site of pPBI96-23 to produce the plasmids pUSN99-1 and pUSN99-2 (FIGS. 27 and 28) representing sense and antisense orientations of the SBEII-1 sequences respectively. 
     pPBI97-2BdUN1. pPBI92-2BdUN1 (also sometimes referred to as p97-2BdUN1) comprises a reconstituted ubiquitin regulatory system (referred to hereafter as a modified ubiquitin promoter or a modified ubiquitin regulatory system (mURS)) which lacks the two overlapping ‘consensus heatshock elements’ discussed in EP 0342926 and U.S. Pat. No. 5,614,399. The modified ubiquitin promoter was prepared via PCR amplification of two DNA fragments using maize genomic DNA as template, followed by ligation of the two fragments to produce a single fragment lacking the consensus heatshock (HS) elements. A Kpn1 restriction site was engineered in place of the HS elements. The primers used were designed from sequence information published by Liu et al 1995 (EMBL DNA database accession ZMU29159). To delete the HS elements and to replace with a diagnostic Kpn1 site the ubiquitin promoter and intron sequences were amplified as two fragments using the primer combinations HS1+Ubi3-3 and HS2+Ubi5-2, the sequences of which are given below. Primers Ubi5-2 and Ubi3-3 are homologous to sequences in the sequence published by Liu et al 1995. Primers HS1 and HS2 are homologous to sequences located immediately 3′ and 5′ respectively of the two overlapping HS elements in the ubiquitin promoter as described in EP 0342926 and U.S. Pat. No. 5,361,399. Both of these primers have a Kpn1 tail at their 5′ ends. 
     Primers 
     HS1: 5-ATTAGGTACCGGACTTGCTCCGCTGTCGGC-3 (SEQ ID No: 48) 
     HS2: 5-TATAGGTACCGAGGCAGCGACAGAGATGCC-3 (SEQ ID No: 49) 
     Ubi5-2: 5-AGCTGAATCCGGCGGCATGGC-3 (SEQ ID No: 50) 
     Ubi3-3: 5-TGATAGTCTTGCCAGTCAGGG-3 (SEQ ID No: 51) 
     The amplified products were subcloned into pGEM TEasy (Promega) to produce the plasmids p97-U1 and p97-U2. The full-length (approx. 2 Kb) modified ubiquitin promoter was reconstructed by subcloning the Kpn1-Sac1 fragment from p97-U1 into the Kpn1/Sac1 sites of p97-U2 to produce p97-U3. A partial restriction map of the predicted sequence (SEQ ID No: 52) of the cloned fragment in p97-U3 is presented in FIG.  29 . (The modified ubiquitin promotor (or mURS) is the subject of a copending European Patent Application filed by the present applicants on the same day as the present application, under the reference C1235.01/M). The modified ubiquitin promoter was transferred as a PstI fragment from p97-U3 into plasmid pPBI96-36. The plasmid pBI96-36 (FIG. 30) comprises the GUS-Nos reporter gene fusion under the control of the wild-type ubiquitin promoter (derived from pAHC25) in a pUC plasmid backbone. The promoter replaces the wild-type ubiquitin regulatory system in pPBI96-36 to produce an intermediary plasmid p97-dUG1 (FIG.  31 ). 
     Construction of pPBI97-2BdUN1 
     The Ubi-Nos sequences in pPBI96-23 were transferred as an EcoRI-HindIII fragment into the EcoRI and HindIII sites of p97-2B (plasmid p97-2B is described in European Patent Application No. 97305694.8 published as WO 99/06570) to produce the plasmid p97-2BUbiNos. The modified ubiquitin promoter was purified as a HindIII/SacI fragment from p97-dUG1 (FIG. 31) and transferred into the HindIII and SacI sites of p97-2BUbiNos, replacing the wild-type ubiquitin promoter to produce p97-2BdUbiNos. The NptII sequence in pUN1 was purified as a SacI fragment and transferred into the SacI site of p97-2BdUbiNos to produce pPBI97-2BdUN1 (FIG.  32 ). Following removal of the ampicillin resistance marker using the method as described in WO 99/06570, the resulting plasmid as used for wheat transformation was designated p97-2BdUN1A−. 
     pCaineo 
     pCaiNeo comprises the NptII gene under control of a CaMV35S promoter and maize Adh1 intron. The plasmid is described in Fromm et al 1986. 
     Transformation of Wheat 
     The following plasmid combinations (co-bombardments) have been used in the transformation of wheat plants: 
     
       
         
               
             
               
               
               
             
           
               
                 TABLE 2 
               
             
             
               
                   
               
               
                 Plasmid combinations used in wheat transformation experiments. 
               
             
          
           
               
                   
                 Starch gene construct/s 
                 Selection marker construct 
               
               
                   
                   
               
               
                   
                   
                 pAHC25 
               
               
                   
                 pWXGS+ 
                 pUN1 
               
               
                   
                 pSR97-26A- antisense 
                 pUN1 or 
               
               
                   
                   
                 p97-2BdUN1 
               
               
                   
                 pSR97-29A- sense 
                 p97-2BdUN1 or pCaiNeo 
               
               
                   
                 pSC98-1A- antisense 
                 p97-2BdUN1 
               
               
                   
                 pUSN-1 sense 
                 p97-2BdUN1 
               
               
                   
                 pUSN-2 antisense 
                 p97-2BdUN1 
               
               
                   
                 pUSN-1 sense &amp; pUSN-2 antisense 
                 pUN1 
               
               
                   
                 pSC98-2A- sense 
                 p97-2BdUN1 
               
               
                   
                   
               
             
          
         
       
     
     The wheat transformation methods used and described here are largely based on those described by Barcelo and Lazzeri, 1995. 
     Embryo wheat plants of the spring cultivar Bobwhite and the winter cultivar Florida were grown in a glasshouse with 16 hr day length supplemented with lights to maintain a minimum light intensity of 500 umol m− 2 s −1  at 0.5M above flag leaf. Glasshouse temperatures were maintained at 19° C.+/−1° C. during the day and 14° C.+/−1° C. at night. 
     Immature embryos of wheat were harvested from developing grain. The seeds were harvested and embryos were cultured at approximately 12 days after anthesis when the embryos were approximately 1 mm in length. Seeds were first rinsed in 70% ethanol for 5 minutes and then sterilised in a 10% solution of Domestos bleach (Domestos is a Trade Mark) for 15 minutes followed by 6 washes with sterile distilled water. Following removal of the embryonic axis the embryos were placed axis surface face down on agargel (Sigma catalogue no. A-3301) solidified MM1 media. The general recipe for MM1 is given in Appendix 1, and the recipes for the various constituents in Appendix 2. The embryos were maintained in darkness for one to two days at 24° C.+/−1° C. prior to bombardment. 
     The plasmids pAHC25, pCAiNeo, pUN1 and p97-2BdUN1 were used to provide selection markers in the combinations with starch gene constructs as detailed in Table 2. pAHC25 (Christensen and Quail 1996) contains a chimeric Ubi-BAR gene which provides selection of transformants to phosphinothricin, the active ingredient in herbicides BASTA™ and Bialophos (see Block, M.de. et al 1987). The plasmids pCAiNeo (Fromm et al., 1986), pUN1 and p97-2BdUN1 contain chimeric promoter-NptII gene fusions and provide selection of transformants against a range of aminoglycoside antibiotics including kanamycin, neomycin, geneticin and paromycin. 
     Particle bombardments was used to introduce plasmids into plant cells. The following method was used to precipitate plasmid DNA onto 0.6 μm gold particles (BIO-RAD catalogue number 165-2262): A total of 5 μg of plasmid DNA was added to a 50 μl sonicated for one minute suspension of gold particle @ 10 mg/ml) in a 1.5 ml microfuge tube. Following a brief vortex for three seconds 50 μl of a 0.5M solution of calcium chloride and 20 μl of a 0.05M solution of spermidine free base were added to the opposite sides of the microfuge tube lid. The tube contents were mixed together by closing the lid and tapping the calcium chloride and spermidine to the bottom of the tube. Following a vortex for three seconds the suspension was centrifuged at 13,000 rpm for 5 seconds. The supernatant was then removed and the pellet resuspended in 150 μl of absolute ethanol. This requires scraping the gold particles off the inside of the tube using a pipette tip. Following a further three second vortex, the sample was centrifuged again and the pellet resuspended in a total volume of 85 μl in absolute ethanol. The particles were vortexed briefly and sonicated for 5 seconds in a Camlab Trisonic T310 water bath sonicator to ensure fine dispersion. An aliquot of 5 μl of the DNA coated gold particles were placed in the centre of a macrocarrier (BIO-RAD catalogue no. 115-2335) and allowed to dry for 30 mins. Particle bombardment was performed by using a Biolisit™ PDS-1000/He (BIO-RAD Instruments, Hercules Calif.) chamber which is illustrated schematically in FIG. 33, using helium pressure of 650 and 900 psi (rupture discs: BIO-RAD catalogue numbers 165-2327 and 165-2328 respectively). 
     Referring to FIG. 33, the illustrated vacuum chamber comprises a housing  10 , the inner side walls of which include a series of recesses  12  for receiving shelves such as sample shelf  14  shown at the fourth level down from the top of the housing. A rupture disc  16  is supported in a He pressure shock tube  18  near the top of the housing. A support  20 , resting in the second set of recesses  12  down from the top of the housing, carries unit  22  that includes a stopping screen and a number of rings  24 , with 11 rings below the support  20  and 3-4 rings above the support  20 . Macrocarrier  26  is supported at the top of unit  22 . The approximate distance from the rupture disc  16  to the macrocarrier  26  is 25 mm, with the approximate distance from the macrocarrier  26  to the stopping screen being 7 mm, and the approximate distance from the stopping screen to the sample shelf  14  being 67 mm. The top of unit  22  is about 21 mm from the bottom of the shock tube  18 , and the bottom unit  22  is about 31 mm from the top of sample shelf  14 . 
     Immature embryos were bombarded between 1 and 2 days after culture. For bombardment the immature embryos were grouped into a circular area of approximately 1 cm in diameter comprising 20-100 embryos, axis side face down on the MM1 media. The Petri dish (not shown) containing the tissue was placed in the chamber on shelf  14 , on the fourth shelf level down from the top, as illustrated in FIG.  33 . The air in the chamber was then evacuated to a vacuum of 28.5 inches of Hg. The macrocarrier  26  was accelerated with a helium shock wave using rupture membranes that burst when the He pressure in the shock tube  18  reaches 650 or 900 psi. Within 1 hour after bombardment the bombarded embryos were plated on MM1 media at 10 embryos per 9 cm petri dish and then maintained in constant darkness at 24° C. for 2-3 weeks. During this period somatic embryogenic callus was produced on the bombarded embryos. 
     After 2-3 weeks the embryos were transferred onto agar-solidified regeneration media, known as R media, and incubated under 16 hr daylength at 24° C. The general recipe for R media is given in Appendix 1. Embryos were transferred on fresh plates at 2-3 week intervals. The composition of the regeneration media varied depending on which selection regime was to be used. For transformants bombarded with the BAR gene the 3 amino solution was omitted and PPT (phosphinothricin) at 1 mg/L, rising to 3 mg/L over a period of three 2-3 week transfers was used for selection. For selection of transformants using the NptII gene three different regimes were used: 1) Geneticin (GIBCO-BRL catalogue no. 10131-019) was incorporated (at 50 mg/L) immediately on transfer to regeneration media and maintained at 50 mg/L on subsequent transfers to regeneration media. 2) &amp; 3) Embryos were first transferred to regeneration media without selection for 12 days and 2-3 weeks, respectively, and thereafter transferred on to media containing Geneticin at 50 mg/L. After 2-3 passages on regeneration media regenerating shoots were transferred to individual culture tubes containing 15 ml of regeneration media at half salt strength with selection at 3 mg/L PPT or 35 mg/L geneticin depending on whether the BAR gene of NptII gene had been used in the original bombardments. Following root formation the regenerated plants were transferred to soil and the glasshouse. 
     Genomic DNA Isolation and Southern Analyses 
     Southern analyses of primary transformants and progeny material were carried out as follows: Freeze dried leaf tissues were ground briefly in a Kontes™ pestle and mortar, and genomic DNA extracted as described in Fulton et al, 1995. 5 μg of DNA were digested with an appropriate restriction enzyme according to the manufacturers instructions, and electrophoresed overnight on a 1% agarose gel, after which the gel was then photographed, washed and blotted onto Hybond N+™ (Amersham International) according to the method of Southern using standard procedures (Sambrook et al 1989). Following blotting, the filters were air dried, baked at 65° C. for 1-2 hours and UV fixed at 312nm for 2 minutes. 
     Probe preparation and labelling for the Southern analyses of transformed material was carried out as described above. 
     GUS histochemistry was performed essentially as described in Jefferson (1987). 
     Evaluation of the Ubiquitin Promoter for Constitutive Expression of Associated Transgenes 
     The plasmid pAHC25 (Christensen and Quail, 1996) was transformed into wheat as described in previous sections. Transformants were selected on the basis of resistance to phosphinothricin. Southern blot analyses were carried out on the primary transformants to confirm integration of the plasmid sequences (data not shown). GUS histochemical analyses were also carried out and demonstrated that the ubiquitin promoter is capable of mediating high levels of GUS expression in a range of wheat tissues. FIG. 34 A, B, C &amp; D show histochemical localisation of GUS expression in the seed, stem, floral and leaf tissues respectively. Southern blot and GUS histochemical analyses were also carried out on self progeny from primary transformants to confirm that the transformation system used is capable of producing transgenic plants which stably transmit the integrated plasmid sequences to progeny plants. FIG. 35 shows a Southern blot of 26 progeny plants of transformant BW119 which had been transformed with pAHC25. In this example genomic DNA from the progeny plants was digested with the restriction enzyme Sac1 and the blot was probed with the GUS gene coding sequence. The Southern blot results are suggestive of the presence of two independently segregating integration loci, each comprising concatamers of pAHC25 plasmid sequences. 
     Evaluation of the Maize Waxy Promoter for Endosperm-specific Expression of Associated Transgenes 
     The plasmids pWxGS+ and pUN1 were co-transformed into wheat as described in previous sections. Transformants were selected on the basis of resistance to geneticin. Southern blot analyses were carried out on the primary transformants to confirm integration of the plasmid sequences (data not shown). Gus histochemical analyses were also carried out to determine the expression profile mediated by the maize waxy promoter. The majority of the transformants that expressed GUS exhibited expression specifically in endosperm tissue, demonstrating the suitability of this promoter for mediating endosperm expression of associated transgenes. FIG. 36 A &amp; B shows endosperm specific expression of GUS in seeds from two independent transformants. We did not observe GUS expression in pollen grains as was seen by Russell and Fromm (1997), however the construct they used also incorporated the maize hsp 70 intron which may conceivably have influenced expression both quantitatively and qualitatively. 
     Transformation of Wheat with Starch Gene Constructs 
     The various construct combinations detailed in Table 2 were co-transformed into wheat using the procedures as described in previous sections. Transformants were selected on the basis of resistance to geneticin. The primary transformants were confirmed positive by Southern blot analysis. Blots were sequentially probed with an NptII coding sequence probe and a SBE coding region probe. FIG. 37 shows an example of a Southern blot which comprises 22 putative transformants which had been co-bombarded with pSR97-29A− or pSR97-26A− and pUN1 or p97-2BdUN1. Genomic DNAs on this blot had been digested with Sac1. The blot was first probed with the NptII probe. Lanes marked with an asterisk correspond to transformants which give a positive signal with the NptII probe. The blot shown in FIG. 37 was probed with the SBEII-1 1 kb SacI fragment. The Sac1 digest is expected to release a 1 kb SBEII-1 hybridising band from both pSR97-29A− and pSR97-26A− plasmid sequences, and the intensity of this band will vary depending on the copy number of inserted plasmid sequences. As can be seen in FIG. 37 several additional SBEII-1 hybridising bands are also observed. Five of these bands are present in all lanes and result from hybridisation to endogenous wheat SBEII-1 sequences. The additional bands of varying size which are observed in the majority of lanes which show the 1 kb hybridising band most likely result from integration events in which one or more copies of the plasmid had been linearised within the 1 kb SBEII-1 sequence prior to integration. In the example shown in FIG. 37, of the 20 NptII positive plants, 16 were found to be co-transformed with the SBEII-1 sequences, representing a co-transformation efficiency of 80%. 
     Differential Scanning Calorimetry (DSC) 
     When heated, an aqueous suspension of starch in excess water undergoes a co-operative endothermic transition known as gelatinisation, as discussed above, entailing a melting of the starch crystallites. Differential scanning calorimetry (DSC) measures the amount of energy (heat) absorbed or released by a sample as it is heated, cooled or held in a constant (isothermal) temperature. DSC has been widely used to study the gelatinisation and retrogradation of starch. 
     DSC analyses were carried out on single grains or pools of 5 grains from primary transformants generated through transformation using each of the gene construct combinations detailed in Table 2. 
     Two different sample preparation and DSC methodologies were used: 
     Method 1 
     Individual seed samples were crushed and ground using a pestle and mortar. The resulting bran was then separated and samples weighed into 50 μm aluminium DSC pans. Water, three times by weight, was added and the sample pans sealed. Analyses were performed using a Perkin-Elmer DSC-7 Robotic™ system equipped with an Intercooler II™, for sub-ambient conditions. Samples were heated from 25° C. to 80° C. at a heating rate of 5° C. min −1 . Gelatinisation enthalpy, onset and peak and end temperatures were recorded. The thermograms were analysed using the Perkin-Elmer software programs (Thermal Analysis Software 7). Gelatinisation enthalpy is expressed in Joules (J)/gram (g) of sample. 
     Method 2 
     Pools of 5 seeds from a single primary transformant, or single seeds from primary transformants, were milled using a Cemotec 1090™ Sample Mill. The milled sample was then passed through a 250 micron sieve to separate the bran from endosperm. Approximately 5 mg of the sieved samples was then accurately weighed into 50 μl aluminium DSC pans. Water, three times by weight, was added and the sample pans sealed. Analyses were performed using a Perkin-Elmer Pyris 1™ DSC equipped with autosampler and Intracooler IP. Samples were heated from 40° C. to 85° C. at a heating rate of 10° C. per minute. The thermograms were analysed using the Perkin-Elmer software programs (Pyris Software for Windows v 3.5). Gelatinisation enthalpy, onset and peak and end temperatures were recorded. 
     Using method 1, DSC analyses were performed on individual mature grains of primary transformants, transformed with the plasmid combinations pSR97-26A−/pUN1, pSR97-26A−/p97-2BdUN1 and pSR97-29A−/p97-2BdUN1. Data obtained were compared to data from control material which had been transformed with one of the NptII selectable marker plasmids, but did not contain any of the ‘starch’ plasmids. Table 3 summarises the average onset, peak, end and enthalpy values for the selected material. The majority of samples showed similar values to the control material. However, as can be seen from Table 3 onset, peak and end temperatures were higher for a number of the transgenic samples compared to the control material. For example, transformant BW 326 exhibits a 6.7° C., 4.9° C. and 4.6° C. increase in onset, peak and end temperatures (respectively) compared to the control sample. 
     Using method 2 a further series of DSC analyses were carried out on pools of 5 grains from primary transformants, transformed with the plasmid combinations pSC98-1A−/p97-2BdUN1, pUSN-1p97-2BdUN1, pUSN-2/p97-2BdUN1 and pUSN-1/pUSN-2/pUNI. Data obtained were compared to data from control material which had been transformed with one of the NptII selectable marker plasmids, but did not contain any of the ‘starch’ plasmids. Table 4 summaries the onset, peak, end and enthalpy values for the selected pooled samples. In many cases there is evidence that the ‘starch’ transgenic material shows onset, peak and end temperatures which are greater than those observed for the control material. For example, transformant BW727 exhibits a 9.8° C., 8.7° C. and 9.1° C. increase in onset, peak and end temperatures (respectively) compared to the BW control sample 3, and a 7.6° C., 6.8° C. and 7.8° C. increase in onset, peak and end temperatures (respectively) compared to the BW control sample 2. 
     
       
         
               
             
               
               
               
               
               
               
             
           
               
                 TABLE 3 
               
             
             
               
                   
               
               
                 Results of DSC analyses on single grains using method 1. Data shown 
               
               
                 are the averages of between 2 and 6 individual grain samples (T o , T p   
               
               
                 and T f  are onset, peak and end temperatures respectively). 
               
             
          
           
               
                   
                 Line 
                 T o   
                 T p   
                 T f   
                 ΔH 
               
               
                 Plasmid combination 
                 Code 
                 (° C.) 
                 (° C.) 
                 (° C.) 
                 (J/g) 
               
               
                   
               
               
                 BW control sample 1 
                   
                 55.2 
                 59.7 
                 66.5 
                 4.66 
               
               
                 pSR97-26A-/pUN1 
                 BW283 
                 57.1 
                 60.4 
                 65.0 
                 2.12 
               
               
                   
                 BW135 
                 57.2 
                 62.1 
                 68.6 
                 4.86 
               
               
                   
                 BW324 
                 57.8 
                 62.1 
                 69.1 
                 5.33 
               
               
                   
                 BW325 
                 58.4 
                 61.8 
                 68.7 
                 3.90 
               
               
                   
                 BW326 
                 61.9 
                 64.6 
                 71.1 
                 2.46 
               
               
                   
                 BW348 
                 60.7 
                 63.4 
                 69.7 
                 3.76 
               
               
                 pSR97-26A-/p97-2BdUN1 
                 F227 
                 57.4 
                 61.4 
                 67.3 
                 2.65 
               
               
                 pSR97-29A-/p97-2BdUN1 
                 F310 
                 62.1 
                 63.7 
                 69.2 
                 6.75 
               
               
                   
                 F312 
                 59.0 
                 62.3 
                 66.8 
                 1.16 
               
               
                   
                 BW335 
                 56.2 
                 60.8 
                 69.1 
                 4.63 
               
               
                   
                 BW353 
                 59.5 
                 62.7 
                 70.8 
                 3.21 
               
               
                   
                 BW354 
                 55.4 
                 61.7 
                 68.9 
                 4.28 
               
               
                   
                 BW355 
                 57.9 
                 61.5 
                 68.0 
                 3.95 
               
               
                   
                 BW357 
                 55.3 
                 60.6 
                 68.0 
                 3.74 
               
               
                   
                 BW363 
                 56.7 
                 62.5 
                 67.9 
                 1.13 
               
               
                   
                 BW367 
                 59.0 
                 62.5 
                 68.2 
                 2.17 
               
               
                   
                 BW369 
                 57.9 
                 60.9 
                 65.9 
                 1.04 
               
               
                   
                 BW370 
                 53.7 
                 59.4 
                 67.5 
                 6.00 
               
               
                   
                 BW375 
                 57.2 
                 61.5 
                 70.0 
                 4.14 
               
               
                   
                 BW376 
                 54.0 
                 58.1 
                 68.0 
                 3.39 
               
               
                   
                 BW377 
                 53.4 
                 60.9 
                 69.2 
                 2.60 
               
               
                   
                 BW380 
                 54.6 
                 61.6 
                 67.6 
                 2.16 
               
               
                   
                 BW390 
                 56.8 
                 61.2 
                 68.5 
                 1.29 
               
               
                   
                 BW399 
                 57.4 
                 62.7 
                 67.9 
                 1.77 
               
               
                   
                 BW400 
                 60.6 
                 63.6 
                 68.1 
                 0.64 
               
               
                   
                 BW341 
                 51.6 
                 59.0 
                 66.4 
                 1.97 
               
               
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
               
               
               
               
             
           
               
                 TABLE 4 
               
             
             
               
                   
               
               
                 Results of DSC analyses on pools of 5 grains using method 2. T o , 
               
               
                 T p  and T f  are onset, peak and end temperatures respectively 
               
             
          
           
               
                   
                 Line 
                 T o   
                 T p   
                 T f   
                 ΔH 
               
               
                 Plasmid combination 
                 Code 
                 (° C.) 
                 (° C.) 
                 (° C.) 
                 (J/g) 
               
               
                   
               
               
                 F control sample 1 
                   
                 60.1 
                 63.9 
                 68.0 
                  6.30 
               
               
                 BW control sample 2 
                   
                 59.3 
                 64.0 
                 68.4 
                  5.94 
               
               
                 BW control sample 3 
                   
                  57.08 
                  62.09 
                  67.08 
                  4.28 
               
               
                 pSC98-1A-/p97-2BdUN1 
                 BW449 
                 59.3 
                 62.9 
                 67.9 
                  3.95 
               
               
                   
                 BW477 
                 57.7 
                 63.6 
                 70.6 
                  8.30 
               
               
                   
                 F492 
                 62.3 
                 66.4 
                 70.2 
                  7.60 
               
               
                   
                 F494 
                 63.6 
                 67.3 
                 71.0 
                  5.73 
               
               
                   
                 BW511 
                 59.6 
                 63.8 
                 67.2 
                  0.98 
               
               
                   
                 BW518 
                 60.2 
                 64.9 
                 69.2 
                  3.57 
               
               
                   
                 BW519 
                 58.4 
                 63.6 
                 68.5 
                  4.13 
               
               
                   
                 BW527 
                 58.7 
                 63.7 
                 69.0 
                  6.38 
               
               
                   
                 BW549 
                 59.9 
                 64.8 
                 69.3 
                  4.48 
               
               
                   
                 BW550 
                 60.2 
                 64.6 
                 68.9 
                  5.06 
               
               
                   
                 BW552 
                 60.8 
                 62.9 
                 67.9 
                  3.74 
               
               
                   
                 BW553 
                 59.5 
                 63.9 
                 67.5 
                  3.60 
               
               
                   
                 BW555 
                 61.0 
                 66.1 
                 68.2 
                  5.43 
               
               
                   
                 BW557 
                 62.7 
                 66.9 
                 71.0 
                  5.08 
               
               
                   
                 BW559 
                 61.6 
                 65.9 
                 70.8 
                  5.08 
               
               
                   
                 BW563 
                 61.4 
                 65.1 
                 69.4 
                  1.90 
               
               
                   
                 BW564 
                 59.4 
                 64.5 
                 73.2 
                  7.08 
               
               
                   
                 BW576 
                 61.8 
                 65.6 
                 69.3 
                  2.65 
               
               
                   
                 BW587 
                 61.3 
                 65.4 
                 69.4 
                  5.36 
               
               
                   
                 BW614 
                 63.9 
                 67.9 
                 71.8 
                  5.83 
               
               
                   
                 BW618 
                 61.3 
                 65.6 
                 69.7 
                  3.54 
               
               
                   
                 BW583a 
                 58.9 
                 63.7 
                 68.0 
                  3.54 
               
               
                   
                 BW631 
                 61.5 
                 65.6 
                 69.7 
                  4.52 
               
               
                   
                 BW633 
                 61.9 
                 66.0 
                 70.2 
                  5.12 
               
               
                   
                 BW634a 
                 60.8 
                 64.9 
                 70.2 
                  5.10 
               
               
                   
                 BW637a 
                 62.8 
                 67.2 
                 72.0 
                  5.16 
               
               
                   
                 BW639 
                 61.8 
                 65.1 
                 68.9 
                  2.15 
               
               
                   
                 BW640a 
                 62.2 
                 66.7 
                 71.0 
                  3.23 
               
               
                   
                 BW642 
                 63.2 
                 67.2 
                 70.9 
                  4.90 
               
               
                   
                 BW698 
                 62.9 
                 67.0 
                 70.9 
                  4.48 
               
               
                   
                 BW700a 
                 63.8 
                 67.6 
                 71.2 
                  3.41 
               
               
                   
                 BE524a 
                 59.4 
                 64.3 
                 68.9 
                  4.05 
               
               
                 pUSN-1/p97-2BdUN1 
                 BW622 
                 59.0 
                 64.1 
                 68.7 
                  4.32 
               
               
                   
                 BW628 
                 56.2 
                 63.3 
                 66.0 
                  6.09 
               
               
                   
                 BW645 
                 57.5 
                 65.6 
                 69.5 
                  5.97 
               
               
                   
                 BW646 
                 61.6 
                 66.4 
                 67.7 
                  3.99 
               
               
                   
                 BW647 
                 61.3 
                 65.4 
                 69.0 
                  3.47 
               
               
                   
                 BW648 
                 59.8 
                 64.4 
                 68.8 
                  4.65 
               
               
                   
                 BW649 
                 61.3 
                 65.6 
                 70.1 
                  5.07 
               
               
                   
                 BW656 
                 59.9 
                 64.6 
                 69.2 
                  5.38 
               
               
                   
                 BW660 
                 62.0 
                 67.3 
                 71.0 
                  4.23 
               
               
                   
                 BW661 
                 61.5 
                 65.8 
                 69.6 
                  3.88 
               
               
                   
                 BW664 
                 61.1 
                 66.1 
                 70.8 
                  4.81 
               
               
                   
                 BW665 
                 61.6 
                 66.5 
                 69.4 
                  5.25 
               
               
                   
                 BW667 
                 63.0 
                 67.1 
                 70.8 
                  3.91 
               
               
                   
                 BW672 
                 63.0 
                 68.1 
                 71.9 
                  5.43 
               
               
                   
                 BW673A 
                 63.1 
                 67.7 
                 71.6 
                  4.83 
               
               
                   
                 BW675 
                 62.1 
                 66.4 
                 71.3 
                 10.97 
               
               
                   
                 BW676 
                 59.8 
                 67.3 
                 71.2 
                  4.21 
               
               
                   
                 BW678 
                 63.0 
                 66.3 
                 69.3 
                  1.20 
               
               
                   
                 BW680 
                 60.8 
                 65.3 
                 70.1 
                  4.94 
               
               
                   
                 BW701 
                 62.3 
                 67.5 
                 72.2 
                  4.70 
               
               
                   
                 BW706 
                 63.0 
                 67.3 
                 71.3 
                  4.94 
               
               
                   
                 BW707 
                 60.9 
                 65.8 
                 70.0 
                  4.77 
               
               
                   
                 BW705 
                 61.7 
                 65.5 
                 68.8 
                  6.11 
               
               
                   
                 BW726 
                 62.6 
                 67.5 
                 71.3 
                  5.44 
               
               
                   
                 BW755 
                 60.8 
                 65.8 
                 70.6 
                  5.18 
               
               
                   
                 BW702 
                 61.9 
                 67.0 
                 71.0 
                  4.44 
               
               
                   
                 BW756 
                 62.3 
                 66.1 
                 69.7 
                  4.83 
               
               
                 pUSN-2/p97-2BdUN1 
                 BW625 
                 62.7 
                 68.2 
                 73.8 
                  4.27 
               
               
                   
                 BW653 
                 60.4 
                 65.3 
                 70.1 
                  6.52 
               
               
                   
                 BW704 
                 60.9 
                 66.2 
                 70.2 
                  4.19 
               
               
                   
                 BW718 
                 61.3 
                 66.9 
                 71.2 
                  4.15 
               
               
                   
                 BW719 
                 62.2 
                 67.2 
                 71.7 
                  5.32 
               
               
                   
                 BW722 
                 64.8 
                 67.5 
                 70.0 
                  2.14 
               
               
                   
                 BW740 
                 63.4 
                 67.9 
                 72.3 
                  5.67 
               
               
                   
                 BW741 
                 62.6 
                 66.9 
                 70.5 
                  5.30 
               
               
                   
                 BW742 
                 64.6 
                 67.9 
                 72.0 
                  6.66 
               
               
                   
                 BW752 
                 62.3 
                 66.3 
                 70.0 
                  4.63 
               
               
                 pUSN-1/pUSN-2/pUN1 
                 BW685 
                 62.6 
                 65.5 
                 69.0 
                  2.60 
               
               
                   
                 BW686A 
                 61.9 
                 66.3 
                 70.2 
                  4.45 
               
               
                   
                 BW714 
                 63.0 
                 67.6 
                 71.3 
                  3.53 
               
               
                   
                 BW727 
                 66.9 
                 70.8 
                 76.2 
                  5.19 
               
               
                   
                 BW728 
                 62.0 
                 66.3 
                 70.4 
                  5.70 
               
               
                   
                 BW731 
                 63.3 
                 67.9 
                 73.0 
                  4.90 
               
               
                   
                 BW732 
                 63.5 
                 66.8 
                 70.8 
                  4.11 
               
               
                   
                 BW748 
                 62.1 
                 67.4 
                 71.9 
                  5.38 
               
               
                   
                 BW794 
                 62.8 
                 67.5 
                 71.8 
                  5.17 
               
               
                   
               
             
          
         
       
     
     Appendix 1 
     Recipe for 2× concentrated MM1 media 
     
       
         
               
               
             
               
               
               
             
           
               
                   
               
               
                   
                 Volume of stock per litre of 2× 
               
               
                 Constituent 
                 concentrated media 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 Macrosalts MS (10× stock) 
                 200 
                 ml 
               
               
                 Microsalts L (1000× stock) 
                 2 
                 ml 
               
               
                 FeNaDTA MS (100× stock) 
                 20 
                 ml 
               
               
                 [Sigma catalogue F-0518] 
               
               
                 Modified Vits MS (×1000) 
                 1 
                 ml 
               
               
                 3 amino acid solution (25× stock) 
                 40 
                 ml 
               
               
                 myo inositol 
                 0.2 
                 g 
               
               
                 (Sigma catalogue number I-3011) 
               
               
                 sucrose 
                 180 
                 g 
               
               
                 AgNO 3  (20 mg/ml stock) 
                 1 
                 ml 
               
               
                 Added after filter sterilisation 
               
               
                 Picloram (1 m/ml stock) 
                 4 
                 ml 
               
               
                 Added after filter sterilisation 
               
               
                   
               
             
          
         
       
     
     Filter sterilise and add to an equal volume of moulten 2× agargel (10 g/L). 
     Recipe for 2× concentrated R media 
     
       
         
               
               
             
               
               
               
             
           
               
                   
               
               
                   
                 Volume of stock per litre of 2× 
               
               
                 Constituent 
                 concentrated media 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 Macrosalts L7 (10× stock) 
                 200 
                 ml 
               
               
                 Microsalts L (1000× stock) 
                 2 
                 ml 
               
               
                 FeNaEDTA MS (100× stock) 
                 20 
                 ml 
               
               
                 Vits/Inositol L2 (200× stock) 
                 10 
                 ml 
               
               
                 3 amino acid solution (25× stock) 
                 40 
                 ml 
               
               
                 Maltose 
                 60 
                 g 
               
               
                 2,4-D (1 mg/ml stock) 
                 200 
                 ml 
               
               
                 added after filter sterilisation 
               
               
                 Zeatin cis trans mi× ed isomers 
                 2 
                 ml 
               
               
                 (Melford labs catalogue no. 
               
               
                 Z-0917) (5 mg/ml stock) added 
               
               
                 after filter sterilisation 
               
               
                   
               
             
          
         
       
     
     Filter sterilise and add to an equal volume of moulten 2× agar (16 g/liter) 
     Appendix 2 
     Recipes for constituents of MM1 and R media 
     Microsalts L (1000× stock) 
     
       
         
               
               
             
               
               
               
               
             
           
               
                   
                   
               
               
                   
                 per 100 ml 
               
               
                   
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 MnSO 4 .7H 2 O 
                 1.34 
                 g 
               
               
                   
                 H 3 BO 3   
                 0.5 
                 g 
               
               
                   
                 ZnSO 4 .7H 2 O 
                 0.75 
                 g 
               
               
                   
                 KI 
                 75 
                 mg 
               
               
                   
                 Na 2 MoO 4 .2H 2 O 
                 25 
                 mg 
               
               
                   
                 CuSO 4 .5H 2 O 
                 2.5 
                 mg 
               
               
                   
                 CoCl 2 .6H 2 O 
                 2.5 
                 mg 
               
               
                   
                   
               
             
          
         
       
     
     Filter sterilise through a 22 μm membrane filter 
     Store at 4° C. 
     Macrosalts MS (10× stock) 
     
       
         
               
               
             
               
               
               
               
             
           
               
                   
                   
               
               
                   
                 per litre 
               
               
                   
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 NH 4 NO 3   
                 16.5 
                 g 
               
               
                   
                 KNO 3   
                 19.0 
                 g 
               
               
                   
                 KH 2 PO 4   
                 1.7 
                 g 
               
               
                   
                 MgSO 4 .7H 2 O 
                 3.7 
                 g 
               
               
                   
                 CaCl 2 .2H 2 O 
                 4.4 
                 g 
               
               
                   
                   
               
             
          
         
       
     
     NB: Dissolve CaCl, before mixing with other components 
     NB: Make up KH 2 PO 4  separately in sterile H 2 0, and add last. 
     Store solution at 4° C. after autoclaving 
     Modified MS Vits (1000× stock) 
     
       
         
               
               
             
               
               
               
               
             
           
               
                   
                   
               
               
                   
                 Per 100 ml 
               
               
                   
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Thiamine HCl 
                 10 
                 mg 
               
               
                   
                 Pyridoxine HCl 
                 50 
                 mg 
               
               
                   
                 Nicotinic acid 
                 50 
                 mg 
               
               
                   
                   
               
             
          
         
       
     
     Store solution in 10 ml aliquots at −20° C. 
     3 amino acid solution (25× stock) 
     
       
         
               
               
             
               
               
               
               
             
           
               
                   
                   
               
               
                   
                 Per litre 
               
               
                   
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 L-Glutamine 
                 18.75 
                 g 
               
               
                   
                 L-Proline 
                 3.75 
                 g 
               
               
                   
                 L-Asparagine 
                 2.5 
                 g 
               
               
                   
                   
               
             
          
         
       
     
     Store solution in 40ml aliquots at −20° C. 
     Macrosalts L7 (10× stock) 
     
       
         
               
               
             
               
               
               
               
             
           
               
                   
                   
               
               
                   
                 per litre 
               
               
                   
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 NH 4 NO 3   
                 2.1 
                 g 
               
               
                   
                 KNO 3   
                 15.0 
                 g 
               
               
                   
                 KH 2 PO 4   
                 2.0 
                 g 
               
               
                   
                 MgSO 4 .7H 2 O 
                 3.5 
                 g 
               
               
                   
                 CaCl 2 .H 2 O 
                 4.5 
                 g 
               
               
                   
                   
               
             
          
         
       
     
     NB: Dissolve CaCl 2  before mixing with other components 
     NB: Make up KH2PO 4  separately in 50 ml H 2 0 and add last 
     Store solution at 4° C. after autoclaving 
     Vits/Insitol (200× stock) 
     
       
         
               
               
               
             
               
               
               
               
             
           
               
                   
                   
               
               
                   
                 200× Stock 
                 Per 100 ml 
               
               
                   
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Inositol 
                 4.0 
                 g 
               
               
                   
                 Thiamine HCl 
                 0.2 
                 g 
               
               
                   
                 Pyridoxine HCl 
                 0.02 
                 g 
               
               
                   
                 Nicotinic acid 
                 0.02 
                 g 
               
               
                   
                 Ca-pantothenate 
                 0.02 
                 g 
               
               
                   
                 Ascorbic acid 
                 0.02 
                 g 
               
               
                   
                   
               
             
          
         
       
     
     Store solution in 40 ml aliquots at −20° C. 
     REFERENCES 
     Atwell, W. A., Hood, L. F., Lineback, D. R., Varriano-Marston, E., and Zobel, H. F. (1988). The terminology and methodology associated with basic starch penomena. Cereal Foods World 33, 306-311. 
     Baba, T., Kimura, K., Mizuno, K., Etoh, H., Ishida, Y., Shida, O., and Arai, Y. (1991). Sequence conservation of the catalytic regions of amylolytic enzymes in maize branching enzyme-I. Biochem. Biophys. Res. Comm. 181(1): 87-94. 
     Barcelo P., and Lazzeri, P. (1995). Transformation of cereals by microprojectile bombardment of immature inflorescence and scutellum tissues. Methods in Molecular Biology—Plant Gene Transfer and Expression Protocols (vol 49), 113-123. Jones H [ed] Humana Press Inc., Totowa, N.J. 
     Block, M.de et al. (1987). EMBO J 6:2513-2518 
     Boyer, C. D. and Priess, J. (1978). Multiple forms of (1-4)-α-D-Glucan, (1-4)-α-D-Glucan 6-glycosyl transferase from developing  Zea mays  L. kernals. Carbohydrate Res 61:321-334. 
     Burton, R. A., Bewley, J. D., Smith, A. M., Bhattacharyya, M. K., Tatge, H., Ring, S., Bull, V., Hamilton, W. D. O., and Martin, C. (1995). Starch branching enzymes belonging to distinct enzyme families are differentially expressed during pea embryo development. Plant J 7:1-13 
     Christensen, A. H., and Quail, P. H. (1996). Transgenic research. 5:213-218. 
     Donovan, J. W. (1979). Phase transitions of the starch-water system. Biopolym. 18, 263-275. 
     Fisher, D. K., Boyer, C. D., and Hannah, L. C. (1993). Starch branching enzyme II from maize endosperm. Plant Physiol. 102:1045-1046. 
     Fisher, D. K., Gao, M., Kim, K-N., Boyer, C. D., and Guiltinan, M. J. (1996). Two closely related cDNAs encoding starch branching enzyme from  Arabidopis thaliana.  Plant Mol. Biol. 30:97-108 
     Fromm M E, Taylor L P &amp; Walbot V. (1986). Stable transformation of maize after gene transfer by electroporation. Nature 319: 791-793, 
     Fulton, T. M., Chunwongse, J., and Tanksley, S. D. (1995). Miniprep Protocol for Extraction of DNA from Tomato and other Herbaceous Plants, Plant Molecular Biology Reporter, 13 (3), pages 225-227. 
     Gao, M., Fisher, D. K., Kim, K-N., Shannon, J. C., and Guiltinan, M. J. (1997). Independent genetic control of maize starch-branching enzymes IIa and IIb: Isolation and characterisation of a Sbe2a cDNA. Plant Physiol. 114: 69-78. 
     Gaun, H. P., and Preiss, J. (1993). Differentiation of the properties of the branching isoenzymes from maize ( Zea mays ). Plant Physiol. 102: 1269-1273. 
     Goldsbrough A P, Belzile F, Yoder J I (1994). Complementation of the tomato anthocyanin without (aw) mutant using the dihydroflavonol 4-reductase gene. Plant Physiology 105, 491-496. 
     Islam, A. K. M. R. (1983). Ditelosomic additions of barley chromosomes to wheat. In Proc. 6th Int. Wheat Genet. Symp.  233-238. Kyoto, Japan.    
     Jack, P. L., Dimitrijevic, T. A. F., and Mayes S. (1994) Assessment of nuclear, mitochondrial and chloroplast RFLP markers in oil palm ( Elaeis guineensis  Jacq.) Theor. Appl. Genet. 90:643-649. 
     Jefferson R A (1987). Assaying chimaeric genes in plants: The GUS gene fusion system. Plant Molecular Biology Reporter 5 (4) 387-405. 
     Kennedy, J. F. and Cabalda, V. M. B. (1991). Bioactive carbohydrates in food. Chemistry in Britain, November, 1017-1019, 1026. 
     Khoshnoodi, J., Ek, B., Rask, L., and Larsson, H. (1993). Characterisation of the 97 and 103 kDa forms of starch branching enzyme from potato tubers. FEBS 332 (1,2):132-138. 
     Kossmann, J., Visser, R. G. F., Muller-Rober, B., Willmitzer, L., and Sonnewald, U. (1991). Cloning and expression anaylsis of a potato cDNA that encodes branching enzyme: evidence for co-expression of starch biosynthetic genes. Mol. Gren. Genet 230: 39-44. 
     Liu et al (1995). Biochem Cell Biol 73: 19-30. 
     Martin, C., and Smith, A. M. (1995). Starch biosynthesis. Plant Cell 7:971-985. 
     Matzke &amp; Matzke (1995). Plant Physiol. 107, 679-685. 
     Mizuno, K., Kawasaki, T., Shimada, H., Satoh, H., Kobayashi, E., Okumura, S., Arai, Y., and Baba, T. (1993). Alteration of the structural properties of starch components by the lack of an isoform of starch branching enzyme in rice seeds. J.Biol. Chem. 268(25): 19084-19091. 
     Morell, M. K., Rahman, S., Abrahams, S. L., and Appels, R. (1995). The biochemistry and molecular biology of starch synthesis in cereals. Auist.J. Plant Physiol. 22: 647-660. 
     Morell, M., Blennow, A., Kosar-Hashemi, B., and Samuel, M. S. (1997). Differential expression and properties of starch branching enzyme isoforms in developing wheat endosperm. Plant Physiol. 113:201-208. 
     Mousley, C. G. (1994). PhD Thesis, Wye College, University of London. 
     Nair, R. B., Baga, M., Scoles, G. J., Kartha, K. K., and Chibbar, R. N. (1997). Isolation, characterisation and expression analysis of a starch branching enzyme II cDNA from wheat. Plant Sci. 122:153-163. 
     Nakamura, Y., Takeichi, T., Kawaguchi, K., and Yamanouchi, H. (1992). Purification of two forms of starch branching enzyme (Q-enzyme) from developing rice endosperm. Physiol. Plant 84: 329-335. 
     Nakamura, Y., and Yamanouchi, H. (1992). Nucleotide sequence of a cDNA encoding starch-branching enzyme, or Q-enzyme I, from rice endosperm. Plant Physiol. 99:1265-1266. 
     Preiss, J. (1991). Biology and molecular biology of starch synthesis and regulation. In Oxford Surveys Plant Mol. Cell Biol. 7:59-114. 
     Rahman, S., Abrahams, S., Abbott, D., Mukai, Y., Samuel, M., Morrell, M., and Appels, R. (1997). A complex arrangement of genes at a starch branching enzyme I locus in the D-genome donor of wheat. Genome 40:465-474. 
     Rapellin, A et al 1997. Isolation of a starch branching enzyme I cDNA from a wheat endosperm library. EMBL database accession Y12320. 
     Russell, D. A. and Fromm, M. E. (1977). Tissue-specific expression in transgenic maize of four endosperm promoters from maize and rice. Transgenic Research 6: 157-168. 
     Sambrook J, Fritsch E F, Maniatis T (1989) Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbour Laboratory Press, Cold Spring Harbour, N.Y. 
     Salehuzzaman, S. N. I. M., Jacobsen, E., and Visser, R. G. F. (1992). Cloning, partial sequencing and expression of a cDNA coding for branching enzyme in cassava. Plant Mol. Biol. 20:809-819. 
     Sears, E. R. (1966). In Chromosome manipulations and plant genetics Edited by R. Riley and K. R. Lavis, Oliver and Boyd, Edinburgh. pp29-45. 
     Sheehy et al (1988) PNAS 85, 9905-8809. 
     Smith, A. M. (1988). Major differences in isoforms of starch-branching enzyme between developing embryos of round- and wrinkled-seeded peas ( Pisum sativum  L.). Planta 175:270-279. 
     Shure, M et al 1983. Cell 35: 225-233. 
     Sun, C., Sathish, P., Deiber, A., and Jansson, C. (1995). Multiple starch branching enzymes in barley endosperm. In Photosynthesis from light to biosphere. P. Mathis (ed.) Vol. V:317-320. 
     Sun, C., Sathish, P., Ahlandsberg, S., and Jansson, C. (1998). The two genes encoding starch-branching enzymes IIa and IIb are differentially expressed in Barley. Plant Physiol. 118:3749 (earliest electronic publication date Sep. 10, 1998). 
     Svegmark, K. and Hermansson, A-M. (1990). Shear induced changes in the viscoelastic behaviour of heat treated potato starch dispersions. Carboyd. Polym. 13, 29-45. 
     Takeda, Y., Gaun, H. P., and Preiss, J. (1993). Branching of amylose by the branching isoenzymes of maize endosperm. Carbohydrate Res. 240:252-263. 
     Van der Krol et al, Mol Gen. Genet. 220, 204-212. 
     
       
         
           
             55 
           
           
             1 
             2307 
             DNA 
             Triticum aestivum 
             
               misc_feature 
               (2036)..(2270) 
               N = any nucleotide 
             
           
            1
catygacggc cagtgacttc gagctcggta cccggggatc cgatttggtg tgtgggagat     60
gttcttgcca aacaatgcag atggttcgcc accaattcct cacggctcac gggtgaaggt    120
gagaatggat actccatctg ggataaagga ttcaattcct gcttggatca agtactccgt    180
gcagactcca ggagatatac catacaatgg aatatattat gatcctcccg aagaggagaa    240
gtatgtattc aagcatcctc aacctaaacg accaaaatca ttgcggatat atgaaacaca    300
tgttggcatg agtagcccgg aaccaaagat caacacatat gcaaacttca gggatgaggt    360
gcttccaaga attaaaagac ttggatacaa tgcagtgcaa ataatggcaa tccaggagca    420
ctcatactat ggaagctttg ggtaccatgt taccaatttc tttgcaccaa gtagccgttt    480
tgggtcccca gaagatttaa aatctttgat tgatagagct cacgagcttg gcttggttgt    540
cctcatggat gttgttcaca gtcacgcgtc aaataatacc ttggacgggt tgaatggttt    600
tgatggcacg gatacacatt acttccatgg cggttcacgg ggccatcact ggatgtggga    660
ttcccgtgtg tttaactatg ggaataagga agttataagg tttctacttt ccaatgcaag    720
atggtggcta gaggagtata agtttgatgg tttccgattc gatggcgcga cctccatgat    780
gtatacccat catggattac aagtaacctt tacaggaagc taccatgaat attttggctt    840
tgccactgat gtagatgcgg tcgtttactt gatgctgatg aatgatctaa ttcatgggtt    900
ttatcctgaa gccgtaacta tcggtgaaga tgttagtgga atgcctacat ttgcccttcc    960
tgttcaagtt ggtggggttg gttttgacta tcgcttacat atggctgttg ccgacaaatg   1020
gattgaactt ctcaaaggaa acgatgaagc ttgggagatg ggtaatattg tgcacacact   1080
aacaaacaga aggtggccgg aaaagtgtgt tacttatgct gaaagtcacg atcaagcact   1140
ggttggagac aagactattg cattctggtt gatggacaag gatatgtatg atttcatggc   1200
tctgaacgga ccttcgacac ctagtattga tcgtggaata gcactgcata aaatgattag   1260
acttatcaca atgggtttag gaggagaggg ttatcttaac tttatgggaa atgagttcgg   1320
gcatcctgaa tggatagact ttccaagagg cccacaagta cttccaactg gtaagttcat   1380
cccaggaaac aacaacagtt acgacaaatg ccgtcgaaga tttgaccagg gtgatgcaga   1440
atttcttagg tatcatggta tgcagcagtt tgatcaggcg atgcagcatc ttgaggaaaa   1500
atatggcttt atgacatcag accaccagta cgtatctcgg aaacatgagg aagataaggt   1560
gatcgtgttt gaaaaagggg acttggtatt tgtgttcaac ttccactgga gtaatagcta   1620
tttcgactac cgggttggct gtttaaagcc tgggaagtac aaggttgtct tagactcaga   1680
cgccggactc tttggtggat ttggtaggat ccatcacact gcagagcact tcacttctga   1740
ctgccaacat gacaacaggc cccattcgtt ctcagtgtac actcctagca gaacctgtgt   1800
tgtctatgct ccaatgaact aaacagcaaa gtgcagcata cgcatgcacg ctgttgttgc   1860
tagcactagc aagaaaaaat cgtatggtca atacaaccag gtgcaaggtt taataagggt   1920
ttgcttcaac gagtcctgga tagacaagac aacatgatga tgtgctctgt gctcccaaat   1980
tcccagggcg ttgtggagaa aaaatgctca tctgtgttat tttatggatc agggangaaa   2040
cctcccccaa anaccccttt tttttttgaa aggnggatag gcccccggtn tctgcatntg   2100
gatgcctcct taaatntttg tagccataaa ccattgctag tgtcctntaa attgacagtt   2160
tagaatagng gttntacttt tgtattttnt ttttgacagt tagactgtat tcctcaaata   2220
atcgacatgt tgtttactcg aagntgagaa ataaaatcag agattgnagn aaaaaaaaaa   2280
aaaaaaaaaa aaaaaaaaaa aaaaaaa                                       2307
 
           
             2 
             758 
             PRT 
             Triticum aestivum 
             
               PEPTIDE 
               (675)..(746) 
               Xaa = any amino acid 
             
           
            2
Ile Asp Gly Gln Leu Arg Ala Arg Tyr Pro Gly Ile Arg Phe Gly Val
1               5                   10                  15
Trp Glu Met Phe Leu Pro Asn Asn Ala Asp Gly Ser Pro Pro Ile Pro
            20                  25                  30
His Gly Ser Arg Val Lys Val Arg Met Asp Thr Pro Ser Gly Ile Lys
        35                  40                  45
Asp Ser Ile Pro Ala Trp Ile Lys Tyr Ser Val Gln Thr Pro Gly Asp
    50                  55                  60
Ile Pro Tyr Asn Gly Ile Tyr Tyr Asp Pro Pro Glu Glu Glu Lys Tyr
65                  70                  75                  80
Val Phe Lys His Pro Gln Pro Lys Arg Pro Lys Ser Leu Arg Ile Tyr
                85                  90                  95
Glu Thr His Val Gly Met Ser Ser Pro Glu Pro Lys Ile Asn Thr Tyr
            100                 105                 110
Ala Asn Phe Arg Asp Glu Val Leu Pro Arg Ile Lys Arg Leu Gly Tyr
        115                 120                 125
Asn Ala Val Gln Ile Met Ala Ile Gln Glu His Ser Tyr Tyr Gly Ser
    130                 135                 140
Phe Gly Tyr His Val Thr Asn Phe Phe Ala Pro Ser Ser Arg Phe Gly
145                 150                 155                 160
Ser Pro Glu Asp Leu Lys Ser Leu Ile Asp Arg Ala His Glu Leu Gly
                165                 170                 175
Leu Val Val Leu Met Asp Val Val His Ser His Ala Ser Asn Asn Thr
            180                 185                 190
Leu Asp Gly Leu Asn Gly Phe Asp Gly Thr Asp Thr His Tyr Phe His
        195                 200                 205
Gly Gly Ser Arg Gly His His Trp Met Trp Asp Ser Arg Val Phe Asn
    210                 215                 220
Tyr Gly Asn Lys Glu Val Ile Arg Phe Leu Leu Ser Asn Ala Arg Trp
225                 230                 235                 240
Trp Leu Glu Glu Tyr Lys Phe Asp Gly Phe Arg Phe Asp Gly Ala Thr
                245                 250                 255
Ser Met Met Tyr Thr His His Gly Leu Gln Val Thr Phe Thr Gly Ser
            260                 265                 270
Tyr His Glu Tyr Phe Gly Phe Ala Thr Asp Val Asp Ala Val Val Tyr
        275                 280                 285
Leu Met Leu Met Asn Asp Leu Ile His Gly Phe Tyr Pro Glu Ala Val
    290                 295                 300
Thr Ile Gly Glu Asp Val Ser Gly Met Pro Thr Phe Ala Leu Pro Val
305                 310                 315                 320
Gln Val Gly Gly Val Gly Phe Asp Tyr Arg Leu His Met Ala Val Ala
                325                 330                 335
Asp Lys Trp Ile Glu Leu Leu Lys Gly Asn Asp Glu Ala Trp Glu Met
            340                 345                 350
Gly Asn Ile Val His Thr Leu Thr Asn Arg Arg Trp Pro Glu Lys Cys
        355                 360                 365
Val Thr Tyr Ala Glu Ser His Asp Gln Ala Leu Val Gly Asp Lys Thr
    370                 375                 380
Ile Ala Phe Trp Leu Met Asp Lys Asp Met Tyr Asp Phe Met Ala Leu
385                 390                 395                 400
Asn Gly Pro Ser Thr Pro Ser Ile Asp Arg Gly Ile Ala Leu His Lys
                405                 410                 415
Met Ile Arg Leu Ile Thr Met Gly Leu Gly Gly Glu Gly Tyr Leu Asn
            420                 425                 430
Phe Met Gly Asn Glu Phe Gly His Pro Glu Trp Ile Asp Phe Pro Arg
        435                 440                 445
Gly Pro Gln Val Leu Pro Thr Gly Lys Phe Ile Pro Gly Asn Asn Asn
    450                 455                 460
Ser Tyr Asp Lys Cys Arg Arg Arg Phe Asp Gln Gly Asp Ala Glu Phe
465                 470                 475                 480
Leu Arg Tyr His Gly Met Gln Gln Phe Asp Gln Ala Met Gln His Leu
                485                 490                 495
Glu Glu Lys Tyr Gly Phe Met Thr Ser Asp His Gln Tyr Val Ser Arg
            500                 505                 510
Lys His Glu Glu Asp Lys Val Ile Val Phe Glu Lys Gly Asp Leu Val
        515                 520                 525
Phe Val Phe Asn Phe His Trp Ser Asn Ser Tyr Phe Asp Tyr Arg Val
    530                 535                 540
Gly Cys Leu Lys Pro Gly Lys Tyr Lys Val Val Leu Asp Ser Asp Ala
545                 550                 555                 560
Gly Leu Phe Gly Gly Phe Gly Arg Ile His His Thr Ala Glu His Phe
                565                 570                 575
Thr Ser Asp Cys Gln His Asp Asn Arg Pro His Ser Phe Ser Val Tyr
            580                 585                 590
Thr Pro Ser Arg Thr Cys Val Val Tyr Ala Pro Met Asn Thr Ala Lys
        595                 600                 605
Cys Ser Ile Arg Met His Ala Val Val Ala Ser Thr Ser Lys Lys Lys
    610                 615                 620
Ser Tyr Gly Gln Tyr Asn Gln Val Gln Gly Leu Ile Arg Val Cys Phe
625                 630                 635                 640
Asn Glu Ser Trp Ile Asp Lys Thr Thr Cys Ala Leu Cys Ser Gln Ile
                645                 650                 655
Pro Arg Ala Leu Trp Arg Lys Asn Ala His Leu Cys Tyr Phe Met Asp
            660                 665                 670
Gln Gly Xaa Asn Leu Pro Gln Xaa Pro Leu Phe Phe Leu Lys Gly Gly
        675                 680                 685
Ala Pro Gly Xaa Cys Xaa Trp Met Pro Pro Xaa Phe Val Ala Ile Asn
    690                 695                 700
His Cys Cys Pro Xaa Asn Gln Phe Arg Ile Xaa Val Xaa Leu Leu Tyr
705                 710                 715                 720
Phe Xaa Phe Asp Ser Thr Val Phe Leu Lys Ser Thr Cys Cys Leu Leu
                725                 730                 735
Glu Xaa Glu Lys Asn Gln Arg Leu Xaa Xaa Lys Lys Lys Lys Lys Lys
            740                 745                 750
Lys Lys Lys Lys Lys Asn
        755
 
           
             3 
             1036 
             DNA 
             Triticum aestivum 
             
               misc_feature 
               (77)..(1036) 
               N = any nucleotide 
             
           
            3
atgtatgatt tcatggctct gaacggacct tcgacgccta atattgatcg tggaatagca     60
ctgcataaaa tgattanact tatcacaatg ggtttaggcg gagagggtta tcttaacttt    120
atgggaaatg agttcgggca tcctgaatgg atagactttc caagaggccc acaagtactt    180
ccaagtggta agttcatccc aggaaacagc aacagttacg acaaatgccg tcgaagattt    240
gacctgggtg atgcagaatt tcttaggtat catggtatgc agcagtttga tcaggcaatg    300
cagcatcttg aggaaaaata tggttttatg acatcagacc accagtacgt atctcggaaa    360
cacgaggaag ataaggtgat cgtgtttgaa aaaggggact tggtatttgt gttcaacttc    420
cactggagta atagctattt cgactaccgg gtcggctgtt taaagcctgg gaagtacaag    480
gtggtcttag actcagacgc tggactcttt ggtggatttg gtaggatcca tcacactgca    540
gagcacttca cttctgactg ccaacatgac aacaggcccc attcgttctc agtgtacact    600
cctagcagaa cctgtgttgt ctatgctcca atgaactaac agcaaggtgc agcatacgcg    660
tgcgcgctgt tgttgctagt agcaagaaaa atcgtacggt caatacagcc aggtgcaagg    720
tttaataagg attttttgct tcaacgagtc ctggatagac aagacaacat gatgttgtgg    780
cgtgtgctcc caatccccag ggcgttgtga agaaaacatg ctcatctgtg ttatgatttt    840
atggatcagc gacgaaactt cccccaaata cccatgcctc cttaaatctt tgtggccgta    900
aaccattgct agtgtcctct aaattgacag tttagcatag aggttttact tttgtatctt    960
ctttttgaca gttagacttt attcctcaaa taatcgacca gtcgtttact cgaaaaaaaa   1020
aaaaaaaaaa aaaaan                                                   1036
 
           
             4 
             1087 
             DNA 
             Triticum aestivum 
             
               misc_feature 
               (201)..(857) 
               N = any nucleotide 
             
           
            4
atgtatgatt tcatggctct gaacggacct tcgacaccta atattgatcg tggaatagca     60
ctgcataaaa tgattagact tatcacaatg ggtttaggag gagagggtta tcttaacttt    120
atgggaaatg agttcgggca tcctgaatgg atagactttc caagaggccc acaagtactt    180
ccaactggta agttcatccc nngaaacaac aacagttacg acaaatgccg tcgaaaattt    240
gacctgggtg atgcagaatt tcttaggtat catggtatgc agcagtttga tcaggcgatg    300
cagcatcttg aggaaaaata tggctttatg acatcagacc accagtacgt atctcggaaa    360
catgaggaag ataaggtgat cgtgtttgaa aaaggggact tggtatttgt gttcaacttc    420
cactggagta atagctattt cggctaccgg gttggctgtt taaagcctgg gaagtacaag    480
gttgtcttag actcagacgc cggactcttt ggtggatttg gtaggatcca tcacactgca    540
gagcacttca cttctgactg ccaacatgac aacaggcccc attcgttctc agtgtacact    600
cctagcagaa cctgtgttgt ctatgctcca atgaactaaa cagcaaagtg cagcatacgc    660
atgcacgctg ttgttgctag cactagcaag aaaaaatcgt atggtcaata caaccaggtg    720
caaggtttaa taagggtttt tgcttcaacg agtcctggat agacaagaca acatgatgat    780
gtgctctgtg ctcccaaatt cccagggcgt tgngnggaaa acatgctcat ctgtgttatc    840
attttatgga tcagngngga aacctccccc aaatacccat gcctccttaa acttttgtgg    900
tcctaaacca tggctactat cctctaaatt ggcagtttag catagaggtt ttacttttgt    960
aaattttttt tgacagttaa tagactctat tcctcaaata attgacatgt cctttacaag   1020
aagatgagaa ataaaatcag ggattgaaga atcccaaaag ctaaaaaaaa aaaaaaaaaa   1080
aaaaaaa                                                             1087
 
           
             5 
             1120 
             DNA 
             Triticum aestivum 
             
               misc_feature 
               (802)..(1083) 
               N = any nucleotide 
             
           
            5
atgtatgatt tcatggcgct gaacggacct tcgacgccta atattgatcg tggaatagca     60
ctgcataaaa tgattagact tatcacaatg ggtctaggag gagagggtta tcttaacttt    120
atgggaaatg agttcgggca tcctgaatgg atagactttc caagaggccc acaagtactt    180
ccaagtggta agttcatccc aggaaacaac aacagttacg acaaatgccg tcgaagattt    240
gacctgggtg atgcagaatt tcttaggtat catggtatgc agcagtttga tcaggcaatg    300
cagcatcttg aggaaaaata tggttttatg acatcagacc accagtacgt ttctcggaaa    360
catgaggaag ataaggtgat cgtgtttgaa aaaggggact tggtatttgt gttcaacttc    420
cactggagta gtagctattt cgactaccgg gtcggctgtt taaagcctgg gaagtacaag    480
gtggtcttag actcggacgc tggactcttt ggtggatttg gtaggatcca tcacactgca    540
gagcacttca cttctgactg ccaacatgac aacaggcccc attcattctc agtgtacact    600
cctagcagaa cctgtgttgt ctatgctcca atgaactaac agcaaagtgc agcatacgcg    660
tgcgcgctgt tgttgctagt agcaagaaaa atcgtatggt caatacaacc aggtgcaagg    720
tttaataagg atttttgctt caacgagtcc tggatagaca agacaacatg atgttgtgct    780
gtgtgctccc aatccccagg gngttgtgaa gaaaacatgc tcatctgtgt tattttatgg    840
atcagggang aaacctcccc caaanacccc tttttttttt gaaaggngga taggcccccg    900
gtntctgcat ntggatgcct ccttaaatnt ttgtagccat aaaccattgc tagtgtcctn    960
taaattgaca gtttagaata gnggttntac ttttgtattt tntttttgac agttagactg   1020
tattcctcaa ataatcgaca tgttgtttac tcgaagntga gaaataaaat cagagattgn   1080
agnaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa                         1120
 
           
             6 
             979 
             DNA 
             Triticum aestivum 
             
               misc_feature 
               (763)..(763) 
               N = any nucleotide 
             
           
            6
atatgtatga tttcatggct ctggataggc cttcaactcc tcgcattgat cgtggcatag     60
cattacataa aatgatcagg cttgtcacca tgggtttagg tggtgaaggc tatcttaact    120
tcatgggaaa tgagtttggg catcctgaat ggatagattt tccaagaggc ccacaaactc    180
ttccaaccgg caaagttctc cctggaaata acaatagtta tgataaatgc cgccatagat    240
ttgatcttgg agatgcagat tttcttagat atcgtggtat gcaagagttc gatcaggcaa    300
tgcagcatct tgaggaaaaa tatgggttta tgacatctga gcaccagtat gtttcacgga    360
aacatgagga agataaggtg atcttcttcg aaagaggaga tttggtattt gttttcaact    420
tccactggag caatagcttt tttgactacc gtgttgggtg ttccaagcct gggaagtaca    480
aggtggcctt ggactccgac gatgcactct ttggtggatt cagcaggctt gatcatgatg    540
tcgactactt cacaaccgaa catccgcatg acaacaggcc gcactctttc tcggtgtaca    600
ctccgagcag aactgcggtc gtgtatgccc ttacagagta agaaccagca gcggcttgtt    660
acaaggcaaa gagagaactc cagagagctc gtggatcgtg agcgaagcga cgggcaacgg    720
cgcgaggctg ctccaagcgc catgactggg aggggatcgt gcntcttccc cagatgccag    780
gaggagcaga tggataggta gcttgttggt gagcgctcga aagaaaatgg acgggcctgg    840
gtgtttgttg tgctgcactg aaccctcctc ctatcttgca cattcccggt tgtttttgta    900
catataacta ataattgccc gtgcgcttca acatgaacat ataaatattc taataggtta    960
aaaaaaaaaa aaaaaaaaa                                                 979
 
           
             7 
             212 
             PRT 
             Triticum aestivum 
           
            7
Met Tyr Asp Phe Met Ala Leu Asn Gly Pro Ser Thr Pro Asn Ile Asp
1               5                   10                  15
Arg Gly Ile Ala Leu His Lys Met Ile Arg Leu Ile Thr Met Gly Leu
            20                  25                  30
Gly Gly Glu Gly Tyr Leu Asn Phe Met Gly Asn Glu Phe Gly His Pro
        35                  40                  45
Glu Trp Ile Asp Phe Pro Arg Gly Pro Gln Val Leu Pro Ser Gly Lys
    50                  55                  60
Phe Ile Pro Gly Asn Ser Asn Ser Tyr Asp Lys Cys Arg Arg Arg Phe
65                  70                  75                  80
Asp Leu Gly Asp Ala Glu Phe Leu Arg Tyr His Gly Met Gln Gln Phe
                85                  90                  95
Asp Gln Ala Met Gln His Leu Glu Glu Lys Tyr Gly Phe Met Thr Ser
            100                 105                 110
Asp His Gln Tyr Val Ser Arg Lys His Glu Glu Asp Lys Val Ile Val
        115                 120                 125
Phe Glu Lys Gly Asp Leu Val Phe Val Phe Asn Phe His Trp Ser Asn
    130                 135                 140
Ser Tyr Phe Asp Tyr Arg Val Gly Cys Leu Lys Pro Gly Lys Tyr Lys
145                 150                 155                 160
Val Val Leu Asp Ser Asp Ala Gly Leu Phe Gly Gly Phe Gly Arg Ile
                165                 170                 175
His His Thr Ala Glu His Phe Thr Ser Asp Cys Gln His Asp Asn Arg
            180                 185                 190
Pro His Ser Phe Ser Val Tyr Thr Pro Ser Arg Thr Cys Val Val Tyr
        195                 200                 205
Ala Pro Met Asn
    210
 
           
             8 
             378 
             DNA 
             Triticum aestivum 
           
            8
actaacagca aggtgcagca tacgcgtgcg cgctgttgtt gctagtagca agaaaaatcg     60
tacggtcaat acagccaggt gcaaggttta ataaggattt tttgcttcaa cgagtcctgg    120
atagacaaga caacatgatg ttgtggcgtg tgctcccaat ccccagggcg ttgtgaagaa    180
aacatgctca tctgtgttat gattttatgg atcagcgacg aaacttcccc caaataccca    240
tgcctcctta aatctttgtg gccgtaaacc attgctagtg tcctctaaat tgacagttta    300
gcatagaggt tttacttttg tatcttcttt ttgacagtta gactttattc ctcaaataat    360
cgaccagtcg tttactcg                                                  378
 
           
             9 
             449 
             DNA 
             Triticum aestivum 
             
               misc_feature 
               (169)..(447) 
               N = any nucleotide 
             
           
            9
aactaacagc aaagtgcagc atacgcgtgc gcgctgttgt tgctagtagc aagaaaaatc     60
gtatggtcaa tacaaccagg tgcaaggttt aataaggatt tttgcttcaa cgagtcctgg    120
atagacaaga caacatgatg ttgtgctgtg tgctcccaat ccccagggng ttgtgaagaa    180
aacatgctca tctgtgttat tttatggatc agggangaaa cctcccccaa anaccccttt    240
tttttttgaa aggnggatag gcccccggtn tctgcatntg gatgcctcct taaatntttg    300
tagccataaa ccattgctag tgtcctntaa attgacagtt tagaatagng gttntacttt    360
tgtattttnt ttttgacagt tagactgtat tcctcaaata atcgacatgt tgtttactcg    420
aagntgagaa ataaaatcag agattgnag                                      449
 
           
             10 
             428 
             DNA 
             Triticum aestivum 
             
               misc_feature 
               (178)..(223) 
               N = any nucleotide 
             
           
            10
actaaacagc aaagtgcagc atacgcatgc acgctgttgt tgctagcact agcaagaaaa     60
aatcgtatgg tcaatacaac caggtgcaag gtttaataag ggtttttgct tcaacgagtc    120
ctggatagac aagacaacat gatgatgtgc tctgtgctcc caaattccca gggcgttgng    180
nggaaaacat gctcatctgt gttatcattt tatggatcag ngnggaaacc tcccccaaat    240
acccatgcct ccttaaactt ttgtggtcct aaaccatggc tactatcctc taaattggca    300
gtttagcata gaggttttac ttttgtaaat tttttttgac agttaataga ctctattcct    360
caaataattg acatgtcctt tacaagaaga tgagaaataa aatcagggat tgaagaatcc    420
caaaagct                                                             428
 
           
             11 
             592 
             PRT 
             Triticum aestivum 
           
            11
Phe Gly Val Trp Glu Met Phe Leu Pro Asn Asn Ala Asp Gly Ser Pro
1               5                   10                  15
Pro Ile Pro His Gly Ser Arg Val Lys Val Arg Met Asp Thr Pro Ser
            20                  25                  30
Gly Ile Lys Asp Ser Ile Pro Ala Trp Ile Lys Tyr Ser Val Gln Thr
        35                  40                  45
Pro Gly Asp Ile Pro Tyr Asn Gly Ile Tyr Tyr Asp Pro Pro Glu Glu
    50                  55                  60
Glu Lys Tyr Val Phe Lys His Pro Gln Pro Lys Arg Pro Lys Ser Leu
65                  70                  75                  80
Arg Ile Tyr Glu Thr His Val Gly Met Ser Ser Pro Glu Pro Lys Ile
                85                  90                  95
Asn Thr Tyr Ala Asn Phe Arg Asp Glu Val Leu Pro Arg Ile Lys Arg
            100                 105                 110
Leu Gly Tyr Asn Ala Val Gln Ile Met Ala Ile Gln Glu His Ser Tyr
        115                 120                 125
Tyr Gly Ser Phe Gly Tyr His Val Thr Asn Phe Phe Ala Pro Ser Ser
    130                 135                 140
Arg Phe Gly Ser Pro Glu Asp Leu Lys Ser Leu Ile Asp Arg Ala His
145                 150                 155                 160
Glu Leu Gly Leu Val Val Leu Met Asp Val Val His Ser His Ala Ser
                165                 170                 175
Asn Asn Thr Leu Asp Gly Leu Asn Gly Phe Asp Gly Thr Asp Thr His
            180                 185                 190
Tyr Phe His Gly Gly Ser Arg Gly His His Trp Met Trp Asp Ser Arg
        195                 200                 205
Val Phe Asn Tyr Gly Asn Lys Glu Val Ile Arg Phe Leu Leu Ser Asn
    210                 215                 220
Ala Arg Trp Trp Leu Glu Glu Tyr Lys Phe Asp Gly Phe Arg Phe Asp
225                 230                 235                 240
Gly Ala Thr Ser Met Met Tyr Thr His His Gly Leu Gln Val Thr Phe
                245                 250                 255
Thr Gly Ser Tyr His Glu Tyr Phe Gly Phe Ala Thr Asp Val Asp Ala
            260                 265                 270
Val Val Tyr Leu Met Leu Met Asn Asp Leu Ile His Gly Phe Tyr Pro
        275                 280                 285
Glu Ala Val Thr Ile Gly Glu Asp Val Ser Gly Met Pro Thr Phe Ala
    290                 295                 300
Leu Pro Val Gln Val Gly Gly Val Gly Phe Asp Tyr Arg Leu His Met
305                 310                 315                 320
Ala Val Ala Asp Lys Trp Ile Glu Leu Leu Lys Gly Asn Asp Glu Ala
                325                 330                 335
Trp Glu Met Gly Asn Ile Val His Thr Leu Thr Asn Arg Arg Trp Pro
            340                 345                 350
Glu Lys Cys Val Thr Tyr Ala Glu Ser His Asp Gln Ala Leu Val Gly
        355                 360                 365
Asp Lys Thr Ile Ala Phe Trp Leu Met Asp Lys Asp Met Tyr Asp Phe
    370                 375                 380
Met Ala Leu Asn Gly Pro Ser Thr Pro Ser Ile Asp Arg Gly Ile Ala
385                 390                 395                 400
Leu His Lys Met Ile Arg Leu Ile Thr Met Gly Leu Gly Gly Glu Gly
                405                 410                 415
Tyr Leu Asn Phe Met Gly Asn Glu Phe Gly His Pro Glu Trp Ile Asp
            420                 425                 430
Phe Pro Arg Gly Pro Gln Val Leu Pro Thr Gly Lys Phe Ile Pro Gly
        435                 440                 445
Asn Asn Asn Ser Tyr Asp Lys Cys Arg Arg Arg Phe Asp Gln Gly Asp
    450                 455                 460
Ala Glu Phe Leu Arg Tyr His Gly Met Gln Gln Phe Asp Gln Ala Met
465                 470                 475                 480
Gln His Leu Glu Glu Lys Tyr Gly Phe Met Thr Ser Asp His Gln Tyr
                485                 490                 495
Val Ser Arg Lys His Glu Glu Asp Lys Val Ile Val Phe Glu Lys Gly
            500                 505                 510
Asp Leu Val Phe Val Phe Asn Phe His Trp Ser Asn Ser Tyr Phe Asp
        515                 520                 525
Tyr Arg Val Gly Cys Leu Lys Pro Gly Lys Tyr Lys Val Val Leu Asp
    530                 535                 540
Ser Asp Ala Gly Leu Phe Gly Gly Phe Gly Arg Ile His His Thr Ala
545                 550                 555                 560
Glu His Phe Thr Ser Asp Cys Gln His Asp Asn Arg Pro His Ser Phe
                565                 570                 575
Ser Val Tyr Thr Pro Ser Arg Thr Cys Val Val Tyr Ala Pro Met Asn
            580                 585                 590
 
           
             12 
             771 
             PRT 
             Triticum aestivum 
           
            12
Ser Arg Ala Ala Ser Pro Gly Lys Val Leu Val Pro Asp Gly Glu Ser
1               5                   10                  15
Asp Asp Leu Ala Ser Pro Ala Gln Pro Glu Glu Leu Gln Ile Pro Glu
            20                  25                  30
Asp Ile Glu Glu Gln Thr Ala Glu Val Asn Met Thr Gly Gly Thr Ala
        35                  40                  45
Glu Lys Leu Glu Ser Ser Glu Pro Thr Gln Gly Ile Val Glu Thr Ile
    50                  55                  60
Thr Asp Gly Val Thr Lys Gly Val Lys Glu Leu Val Val Gly Glu Lys
65                  70                  75                  80
Pro Arg Val Val Pro Lys Pro Gly Asp Gly Gln Lys Ile Tyr Glu Ile
                85                  90                  95
Asp Pro Thr Leu Lys Asp Phe Arg Ser His Leu Asp Tyr Arg Tyr Ser
            100                 105                 110
Glu Tyr Arg Arg Ile Arg Ala Ala Ile Asp Gln His Glu Gly Gly Leu
        115                 120                 125
Glu Ala Phe Ser Arg Gly Tyr Glu Lys Leu Gly Phe Thr Arg Ser Ala
    130                 135                 140
Glu Gly Ile Thr Tyr Arg Glu Trp Ala Pro Gly Ala His Ser Ala Ala
145                 150                 155                 160
Leu Val Gly Asp Phe Asn Asn Trp Asn Pro Asn Ala Asp Thr Met Thr
                165                 170                 175
Arg Asp Asp Tyr Gly Val Trp Glu Ile Phe Leu Pro Asn Asn Ala Asp
            180                 185                 190
Gly Ser Pro Ala Ile Pro His Gly Ser Arg Val Lys Ile Arg Met Asp
        195                 200                 205
Thr Pro Ser Gly Val Lys Asp Ser Ile Ser Ala Trp Ile Lys Phe Ser
    210                 215                 220
Val Gln Ala Pro Gly Glu Ile Pro Phe Asn Gly Ile Tyr Tyr Asp Pro
225                 230                 235                 240
Pro Glu Glu Glu Lys Tyr Val Phe Gln His Pro Gln Pro Lys Arg Pro
                245                 250                 255
Glu Ser Leu Arg Ile Tyr Glu Ser His Ile Gly Met Ser Ser Pro Glu
            260                 265                 270
Pro Lys Ile Asn Ser Tyr Ala Asn Phe Arg Asp Glu Val Leu Pro Arg
        275                 280                 285
Ile Lys Arg Leu Gly Tyr Asn Ala Val Gln Ile Met Ala Ile Gln Glu
    290                 295                 300
His Ser Tyr Tyr Ala Ser Phe Gly Tyr His Val Thr Asn Phe Phe Ala
305                 310                 315                 320
Pro Ser Ser Arg Phe Gly Thr Pro Glu Asp Leu Lys Ser Leu Ile Asp
                325                 330                 335
Arg Ala His Glu Leu Gly Leu Ile Val Leu Met Asp Ile Val His Ser
            340                 345                 350
His Ser Ser Asn Asn Thr Leu Asp Gly Leu Asn Gly Phe Asp Gly Thr
        355                 360                 365
Asp Thr His Tyr Phe His Gly Gly Pro Arg Gly His His Trp Met Trp
    370                 375                 380
Asp Ser Arg Leu Phe Asn Tyr Gly Ser Trp Glu Val Leu Arg Phe Leu
385                 390                 395                 400
Leu Ser Asn Ala Arg Trp Trp Leu Glu Glu Tyr Lys Phe Asp Gly Phe
                405                 410                 415
Arg Phe Asp Gly Val Thr Ser Met Met Tyr Thr His His Gly Leu Gln
            420                 425                 430
Met Thr Phe Thr Gly Asn Tyr Gly Glu Tyr Phe Gly Phe Ala Thr Asp
        435                 440                 445
Val Asp Ala Val Val Tyr Leu Met Leu Val Asn Asp Leu Ile His Gly
    450                 455                 460
Leu His Pro Asp Ala Val Ser Ile Gly Glu Asp Val Ser Gly Met Pro
465                 470                 475                 480
Thr Phe Cys Ile Pro Val Pro Asp Gly Gly Val Gly Leu Asp Tyr Arg
                485                 490                 495
Leu His Met Ala Val Ala Asp Lys Trp Ile Glu Leu Leu Lys Gln Ser
            500                 505                 510
Asp Glu Ser Trp Lys Met Gly Asp Ile Val His Thr Leu Thr Asn Arg
        515                 520                 525
Arg Trp Leu Glu Lys Cys Val Thr Tyr Ala Glu Ser His Asp Gln Ala
    530                 535                 540
Leu Val Gly Asp Lys Thr Ile Ala Phe Trp Leu Met Asp Lys Asp Met
545                 550                 555                 560
Tyr Asp Phe Met Ala Leu Asp Arg Pro Ser Thr Pro Arg Ile Asp Arg
                565                 570                 575
Gly Ile Ala Leu His Lys Met Ile Arg Leu Val Thr Met Gly Leu Gly
            580                 585                 590
Gly Glu Gly Tyr Leu Asn Phe Met Gly Asn Glu Phe Gly His Pro Glu
        595                 600                 605
Trp Ile Asp Phe Pro Arg Gly Pro Gln Thr Leu Pro Thr Gly Lys Val
    610                 615                 620
Leu Pro Gly Asn Asn Asn Ser Tyr Asp Lys Cys Arg Arg Arg Phe Asp
625                 630                 635                 640
Leu Gly Asp Ala Asp Phe Leu Arg Tyr His Gly Met Gln Glu Phe Asp
                645                 650                 655
Gln Ala Met Gln His Leu Glu Glu Lys Tyr Gly Phe Met Thr Ser Glu
            660                 665                 670
His Gln Tyr Val Ser Arg Lys His Glu Glu Asp Lys Val Ile Ile Phe
        675                 680                 685
Glu Arg Gly Asp Leu Val Phe Val Phe Asn Phe His Trp Ser Asn Ser
    690                 695                 700
Phe Phe Asp Tyr Arg Val Gly Cys Ser Arg Pro Gly Lys Tyr Lys Val
705                 710                 715                 720
Ala Leu Asp Ser Asp Asp Ala Leu Phe Gly Gly Phe Ser Arg Leu Asp
                725                 730                 735
His Asp Val Asp Tyr Phe Thr Thr Glu His Pro His Asp Asn Arg Pro
            740                 745                 750
Arg Ser Phe Ser Val Tyr Thr Pro Ser Arg Thr Ala Val Val Tyr Ala
        755                 760                 765
Leu Thr Glu
    770
 
           
             13 
             797 
             PRT 
             Zea mays 
           
            13
Ser Cys Ala Gly Ala Pro Gly Lys Val Leu Val Pro Gly Gly Gly Ser
1               5                   10                  15
Asp Asp Leu Leu Ser Ser Ala Glu Pro Val Val Asp Thr Gln Pro Glu
            20                  25                  30
Glu Leu Gln Ile Pro Glu Ala Glu Leu Thr Val Glu Lys Thr Ser Ser
        35                  40                  45
Ser Pro Thr Gln Thr Thr Ser Ala Val Ala Glu Ala Ser Ser Gly Val
    50                  55                  60
Glu Ala Glu Glu Arg Pro Glu Leu Ser Ser Glu Val Ile Gly Val Gly
65                  70                  75                  80
Gly Thr Gly Gly Thr Lys Ile Asp Gly Ala Gly Ile Lys Ala Lys Ala
                85                  90                  95
Pro Leu Val Glu Glu Lys Pro Arg Val Ile Pro Pro Pro Gly Asp Gly
            100                 105                 110
Gln Arg Ile Tyr Glu Ile Asp Pro Met Leu Glu Gly Phe Arg Gly His
        115                 120                 125
Leu Asp Tyr Arg Tyr Ser Glu Tyr Lys Arg Leu Arg Ala Ala Ile Asp
    130                 135                 140
Gln His Glu Gly Gly Leu Asp Ala Phe Ser Arg Gly Tyr Glu Lys Leu
145                 150                 155                 160
Gly Phe Thr Arg Ser Ala Glu Gly Ile Thr Tyr Arg Glu Trp Ala Pro
                165                 170                 175
Gly Ala Tyr Ser Ala Ala Leu Val Gly Asp Phe Asn Asn Trp Asn Pro
            180                 185                 190
Asn Ala Asp Ala Met Ala Arg Asn Glu Tyr Gly Val Trp Glu Ile Phe
        195                 200                 205
Leu Pro Asn Asn Ala Asp Gly Ser Pro Ala Ile Pro His Gly Ser Arg
    210                 215                 220
Val Lys Ile Arg Met Asp Thr Pro Ser Gly Val Lys Asp Ser Ile Pro
225                 230                 235                 240
Ala Trp Ile Lys Phe Ser Val Gln Ala Pro Gly Glu Ile Pro Tyr Asn
                245                 250                 255
Gly Ile Tyr Tyr Asp Pro Pro Glu Glu Glu Lys Tyr Val Phe Lys His
            260                 265                 270
Pro Gln Pro Lys Arg Pro Lys Ser Leu Arg Ile Tyr Glu Ser His Val
        275                 280                 285
Gly Met Ser Ser Pro Glu Pro Lys Ile Asn Thr Tyr Ala Asn Phe Arg
    290                 295                 300
Asp Glu Val Leu Pro Arg Ile Lys Lys Leu Gly Tyr Asn Ala Val Gln
305                 310                 315                 320
Ile Met Ala Ile Gln Glu His Ser Tyr Tyr Ala Ser Phe Gly Tyr His
                325                 330                 335
Val Thr Asn Phe Phe Ala Pro Ser Ser Arg Phe Gly Thr Pro Glu Asp
            340                 345                 350
Leu Lys Ser Leu Ile Asp Lys Ala His Glu Leu Gly Leu Leu Val Leu
        355                 360                 365
Met Asp Ile Val His Ser His Ser Ser Asn Asn Thr Leu Asp Gly Leu
    370                 375                 380
Asn Gly Phe Asp Gly Thr Asp Thr His Tyr Phe His Gly Gly Pro Arg
385                 390                 395                 400
Gly His His Trp Met Trp Asp Ser Arg Leu Phe Asn Tyr Gly Ser Trp
                405                 410                 415
Glu Val Leu Arg Phe Leu Leu Ser Asn Ala Arg Trp Trp Leu Glu Glu
            420                 425                 430
Tyr Lys Phe Asp Gly Phe Arg Phe Asp Gly Val Thr Ser Met Met Tyr
        435                 440                 445
Thr His His Gly Leu Gln Val Thr Phe Thr Gly Asn Tyr Gly Glu Tyr
    450                 455                 460
Phe Gly Phe Ala Thr Asp Val Asp Ala Val Val Tyr Leu Met Leu Val
465                 470                 475                 480
Asn Asp Leu Ile Arg Gly Leu Tyr Pro Glu Ala Val Ser Ile Gly Glu
                485                 490                 495
Asp Val Ser Gly Met Pro Thr Phe Cys Ile Pro Val Gln Asp Gly Gly
            500                 505                 510
Val Gly Phe Asp Tyr Arg Leu His Met Ala Val Pro Asp Lys Trp Ile
        515                 520                 525
Glu Leu Leu Lys Gln Ser Asp Glu Tyr Trp Glu Met Gly Asp Ile Val
    530                 535                 540
His Thr Leu Thr Asn Arg Arg Trp Leu Glu Lys Cys Val Thr Tyr Cys
545                 550                 555                 560
Glu Ser His Asp Gln Ala Leu Val Gly Asp Lys Thr Ile Ala Phe Trp
                565                 570                 575
Leu Met Asp Lys Asp Met Tyr Asp Phe Met Ala Leu Asp Arg Pro Ser
            580                 585                 590
Thr Pro Arg Ile Asp Arg Gly Ile Ala Leu His Lys Met Ile Arg Leu
        595                 600                 605
Val Thr Met Gly Leu Gly Gly Glu Gly Tyr Leu Asn Phe Met Gly Asn
    610                 615                 620
Glu Phe Gly His Pro Glu Trp Ile Asp Phe Pro Arg Gly Pro Gln Ser
625                 630                 635                 640
Leu Pro Asn Gly Ser Val Ile Pro Gly Asn Asn Asn Ser Phe Asp Lys
                645                 650                 655
Cys Arg Arg Arg Phe Asp Leu Gly Asp Ala Asp Tyr Leu Arg Tyr Arg
            660                 665                 670
Gly Met Gln Glu Phe Asp Gln Ala Met Gln His Leu Glu Gly Lys Tyr
        675                 680                 685
Glu Phe Met Thr Ser Asp His Ser Tyr Val Ser Arg Lys His Glu Glu
    690                 695                 700
Asp Lys Val Ile Ile Phe Glu Arg Gly Asp Leu Val Phe Val Phe Asn
705                 710                 715                 720
Phe His Trp Ser Asn Ser Tyr Phe Asp Tyr Arg Val Gly Cys Phe Lys
                725                 730                 735
Pro Gly Lys Tyr Lys Ile Val Leu Asp Ser Asp Asp Gly Leu Phe Gly
            740                 745                 750
Gly Phe Ser Arg Leu Asp His Asp Ala Glu Tyr Phe Thr Ala Asp Trp
        755                 760                 765
Pro His Asp Asn Arg Pro Cys Ser Phe Ser Val Tyr Ala Pro Ser Arg
    770                 775                 780
Thr Ala Val Val Tyr Ala Pro Ala Gly Ala Glu Asp Glu
785                 790                 795
 
           
             14 
             747 
             PRT 
             Zea mays 
           
            14
Ala Ala Ala Ala Ala Arg Lys Ala Val Met Val Pro Glu Gly Glu Asn
1               5                   10                  15
Asp Gly Leu Ala Ser Arg Ala Asp Ser Ala Gln Phe Gln Ser Asp Glu
            20                  25                  30
Leu Glu Val Pro Asp Ile Ser Glu Glu Thr Thr Cys Gly Ala Gly Val
        35                  40                  45
Ala Asp Ala Gln Ala Leu Asn Arg Val Arg Val Val Pro Pro Pro Ser
    50                  55                  60
Asp Gly Gln Lys Ile Phe Gln Ile Asp Pro Met Leu Gln Gly Tyr Lys
65                  70                  75                  80
Tyr His Leu Glu Tyr Arg Tyr Ser Leu Tyr Arg Arg Ile Arg Ser Asp
                85                  90                  95
Ile Asp Glu His Glu Gly Gly Leu Glu Ala Phe Ser Arg Ser Tyr Glu
            100                 105                 110
Lys Phe Gly Phe Asn Ala Ser Ala Glu Gly Ile Thr Tyr Arg Glu Trp
        115                 120                 125
Ala Pro Gly Ala Phe Ser Ala Ala Leu Val Gly Asp Val Asn Asn Trp
    130                 135                 140
Asp Pro Asn Ala Asp Arg Met Ser Lys Asn Glu Phe Gly Val Trp Glu
145                 150                 155                 160
Ile Phe Leu Pro Asn Asn Ala Asp Gly Thr Ser Pro Ile Pro His Gly
                165                 170                 175
Ser Arg Val Lys Val Arg Met Asp Thr Pro Ser Gly Ile Lys Asp Ser
            180                 185                 190
Ile Pro Ala Trp Ile Lys Tyr Ser Val Gln Ala Pro Gly Glu Ile Pro
        195                 200                 205
Tyr Asp Gly Ile Tyr Tyr Asp Pro Pro Glu Glu Val Lys Tyr Val Phe
    210                 215                 220
Arg His Ala Gln Pro Lys Arg Pro Lys Ser Leu Arg Ile Tyr Glu Thr
225                 230                 235                 240
His Val Gly Met Ser Ser Pro Glu Pro Lys Ile Asn Thr Tyr Val Asn
                245                 250                 255
Phe Arg Asp Glu Val Leu Pro Arg Ile Lys Lys Leu Gly Tyr Asn Ala
            260                 265                 270
Val Gln Ile Met Ala Ile Gln Glu His Ser Tyr Tyr Gly Ser Phe Gly
        275                 280                 285
Tyr His Val Thr Asn Phe Phe Ala Pro Ser Ser Arg Phe Gly Thr Pro
    290                 295                 300
Glu Asp Leu Lys Ser Leu Ile Asp Arg Ala His Glu Leu Gly Leu Leu
305                 310                 315                 320
Val Leu Met Asp Val Val His Ser His Ala Ser Ser Asn Thr Leu Asp
                325                 330                 335
Gly Leu Asn Gly Phe Asp Gly Thr Asp Thr His Tyr Phe His Ser Gly
            340                 345                 350
Pro Arg Gly His His Trp Met Trp Asp Ser Arg Leu Phe Asn Tyr Gly
        355                 360                 365
Asn Trp Glu Val Leu Arg Phe Leu Leu Ser Asn Ala Arg Trp Trp Leu
    370                 375                 380
Glu Glu Tyr Lys Phe Asp Gly Phe Arg Phe Asp Gly Val Thr Ser Met
385                 390                 395                 400
Met Tyr Thr His His Gly Leu Gln Val Thr Phe Thr Gly Asn Phe Asn
                405                 410                 415
Glu Tyr Phe Gly Phe Ala Thr Asp Val Asp Ala Val Val Tyr Leu Met
            420                 425                 430
Leu Val Asn Asp Leu Ile His Gly Leu Tyr Pro Glu Ala Val Thr Ile
        435                 440                 445
Gly Glu Asp Val Ser Gly Met Pro Thr Phe Ala Leu Pro Val His Asp
    450                 455                 460
Gly Gly Val Gly Phe Asp Tyr Arg Met His Met Ala Val Ala Asp Lys
465                 470                 475                 480
Trp Ile Asp Leu Leu Lys Gln Ser Asp Glu Thr Trp Lys Met Gly Asp
                485                 490                 495
Ile Val His Thr Leu Thr Asn Arg Arg Trp Leu Glu Lys Cys Val Thr
            500                 505                 510
Tyr Ala Glu Ser His Asp Gln Ala Leu Val Gly Asp Lys Thr Ile Ala
        515                 520                 525
Phe Trp Leu Met Asp Lys Asp Met Tyr Asp Phe Met Ala Leu Asp Arg
    530                 535                 540
Pro Ser Thr Pro Thr Ile Asp Arg Gly Ile Ala Leu His Lys Met Ile
545                 550                 555                 560
Arg Leu Ile Thr Met Gly Leu Gly Gly Glu Gly Tyr Leu Asn Phe Met
                565                 570                 575
Gly Asn Glu Phe Gly His Pro Glu Trp Ile Asp Phe Pro Arg Gly Pro
            580                 585                 590
Gln Arg Leu Pro Ser Gly Lys Phe Ile Pro Gly Asn Asn Asn Ser Tyr
        595                 600                 605
Asp Lys Cys Arg Arg Arg Phe Asp Leu Gly Asp Ala Asp Tyr Leu Arg
    610                 615                 620
Tyr His Gly Met Gln Glu Phe Asp Gln Ala Met Gln His Leu Glu Gln
625                 630                 635                 640
Lys Tyr Glu Phe Met Thr Ser Asp His Gln Tyr Ile Ser Arg Lys His
                645                 650                 655
Glu Glu Asp Lys Val Ile Val Phe Glu Lys Gly Asp Leu Val Phe Val
            660                 665                 670
Phe Asn Phe His Cys Asn Asn Ser Tyr Phe Asp Tyr Arg Ile Gly Cys
        675                 680                 685
Arg Lys Pro Gly Val Tyr Lys Val Val Leu Asp Ser Asp Ala Gly Leu
    690                 695                 700
Phe Gly Gly Phe Ser Arg Ile His His Ala Ala Glu His Phe Thr Ala
705                 710                 715                 720
Asp Cys Ser His Asp Asn Arg Pro Tyr Ser Phe Ser Val Tyr Thr Pro
                725                 730                 735
Ser Arg Thr Cys Val Val Tyr Ala Pro Val Glu
            740                 745
 
           
             15 
             50 
             PRT 
             Hordeum vulgare 
           
            15
Asn Asp Leu Gly Ile Trp Glu Ile Phe Leu Pro Asn Asn Ala Asp Gly
1               5                   10                  15
Ser Pro Pro Ile Pro His Gly Ser Arg Val Lys Val Arg Met Asp Thr
            20                  25                  30
Pro Ser Gly Thr Lys Asp Ser Ile Pro Ala Trp Ile Lys Phe Ser Val
        35                  40                  45
Gln Ala
    50
 
           
             16 
             50 
             PRT 
             Hordeum vulgare 
           
            16
Asp Asp Tyr Gly Val Trp Glu Ile Phe Leu Pro Asn Asn Ala Asp Gly
1               5                   10                  15
Ser Pro Ala Ile Pro His Gly Ser Arg Val Lys Ile Arg Met Asp Thr
            20                  25                  30
Pro Ser Gly Val Lys Asp Ser Ile Ser Ala Trp Ile Lys Phe Ser Val
        35                  40                  45
Gln Ala
    50
 
           
             17 
             760 
             PRT 
             Oryza sativa 
           
            17
Ala Ala Gly Ala Ser Gly Glu Val Met Ile Pro Glu Gly Glu Ser Asp
1               5                   10                  15
Gly Met Pro Val Ser Ala Gly Ser Asp Asp Leu Gln Leu Pro Ala Leu
            20                  25                  30
Asp Asp Glu Leu Ser Thr Glu Val Gly Ala Glu Val Glu Ile Glu Ser
        35                  40                  45
Ser Gly Ala Ser Asp Val Glu Gly Val Lys Arg Val Val Glu Glu Leu
    50                  55                  60
Ala Ala Glu Gln Lys Pro Arg Val Val Pro Pro Thr Gly Asp Gly Gln
65                  70                  75                  80
Lys Ile Phe Gln Met Asp Ser Met Leu Asn Gly Tyr Lys Tyr His Leu
                85                  90                  95
Glu Tyr Arg Tyr Ser Leu Tyr Arg Arg Leu Arg Ser Asp Ile Asp Gln
            100                 105                 110
Tyr Glu Gly Gly Leu Glu Thr Phe Ser Arg Gly Tyr Glu Lys Phe Gly
        115                 120                 125
Phe Asn His Ser Ala Glu Gly Val Thr Tyr Arg Glu Trp Ala Pro Gly
    130                 135                 140
Ala His Ser Ala Ala Leu Val Gly Asp Phe Asn Asn Trp Asn Pro Asn
145                 150                 155                 160
Ala Asp Arg Met Ser Lys Asn Glu Phe Gly Val Trp Glu Ile Phe Leu
                165                 170                 175
Pro Asn Asn Ala Asp Gly Ser Ser Pro Ile Pro His Gly Ser Arg Val
            180                 185                 190
Lys Val Arg Met Glu Thr Pro Ser Gly Ile Lys Asp Ser Ile Pro Ala
        195                 200                 205
Trp Ile Lys Tyr Ser Val Gln Ala Ala Gly Glu Ile Pro Tyr Asn Gly
    210                 215                 220
Ile Tyr Tyr Asp Pro Pro Glu Glu Glu Lys Tyr Ile Phe Lys His Pro
225                 230                 235                 240
Gln Pro Lys Arg Pro Lys Ser Leu Arg Ile Tyr Glu Thr His Val Gly
                245                 250                 255
Met Ser Ser Thr Glu Pro Lys Ile Asn Thr Tyr Ala Asn Phe Arg Asp
            260                 265                 270
Glu Val Leu Pro Arg Ile Lys Lys Leu Gly Tyr Asn Ala Val Gln Ile
        275                 280                 285
Met Ala Ile Gln Glu His Ala Tyr Tyr Gly Ser Phe Gly Tyr His Val
    290                 295                 300
Thr Asn Phe Phe Ala Pro Ser Ser Arg Phe Gly Thr Pro Glu Asp Leu
305                 310                 315                 320
Lys Ser Leu Ile Asp Lys Ala His Glu Leu Gly Leu Val Val Leu Met
                325                 330                 335
Asp Val Val His Ser His Ala Ser Asn Asn Thr Leu Asp Gly Leu Asn
            340                 345                 350
Gly Phe Asp Gly Thr Asp Thr His Tyr Phe His Ser Gly Ser Arg Gly
        355                 360                 365
His His Trp Met Trp Asp Ser Arg Leu Phe Asn Tyr Gly Asn Trp Glu
    370                 375                 380
Val Leu Arg Phe Leu Leu Ser Asn Ala Arg Trp Trp Leu Glu Glu Tyr
385                 390                 395                 400
Lys Phe Asp Gly Phe Arg Phe Asp Gly Val Thr Ser Met Met Tyr Thr
                405                 410                 415
His His Gly Leu Gln Val Ala Phe Thr Gly Asn Tyr Ser Glu Tyr Phe
            420                 425                 430
Gly Phe Ala Thr Asp Ala Asp Ala Val Val Tyr Leu Met Leu Val Asn
        435                 440                 445
Asp Leu Ile His Gly Leu Tyr Pro Glu Ala Ile Thr Ile Gly Glu Asp
    450                 455                 460
Val Ser Gly Met Pro Thr Phe Ala Leu Pro Val Gln Asp Gly Gly Val
465                 470                 475                 480
Gly Phe Asp Tyr Arg Leu His Met Ala Val Pro Asp Lys Trp Ile Glu
                485                 490                 495
Leu Leu Lys Gln Ser Asp Glu Ser Trp Lys Met Gly Asp Ile Val His
            500                 505                 510
Thr Leu Thr Asn Arg Arg Trp Ser Glu Lys Cys Val Thr Tyr Ala Glu
        515                 520                 525
Ser His Asp Gln Ala Leu Val Gly Asp Lys Thr Ile Ala Phe Trp Leu
    530                 535                 540
Met Asp Lys Asp Met Tyr Asp Phe Met Ala Leu Asp Arg Pro Ala Thr
545                 550                 555                 560
Pro Ser Ile Asp Arg Gly Ile Ala Leu His Lys Met Ile Arg Leu Ile
                565                 570                 575
Thr Met Gly Leu Gly Gly Glu Gly Tyr Leu Asn Phe Met Gly Asn Glu
            580                 585                 590
Phe Gly His Pro Glu Trp Ile Asp Phe Pro Arg Ala Pro Gln Val Leu
        595                 600                 605
Pro Asn Gly Lys Phe Ile Pro Gly Asn Asn Asn Ser Tyr Asp Lys Cys
    610                 615                 620
Arg Arg Arg Phe Asp Leu Gly Asp Ala Asp Tyr Leu Arg Tyr Arg Gly
625                 630                 635                 640
Met Leu Glu Phe Asp Arg Ala Met Gln Ser Leu Glu Glu Lys Tyr Gly
                645                 650                 655
Phe Met Thr Ser Asp His Gln Tyr Ile Ser Arg Lys His Glu Glu Asp
            660                 665                 670
Lys Met Ile Ile Phe Glu Lys Gly Asp Leu Val Phe Val Phe Asn Phe
        675                 680                 685
His Trp Ser Asn Ser Tyr Phe Asp Tyr Arg Val Gly Cys Leu Lys Pro
    690                 695                 700
Gly Lys Tyr Lys Val Val Leu Asp Ser Asp Ala Gly Leu Phe Gly Gly
705                 710                 715                 720
Phe Gly Arg Ile His His Thr Ala Glu His Phe Thr Ala Asp Cys Ser
                725                 730                 735
His Asp Asn Arg Pro Tyr Ser Phe Ser Val Tyr Ser Pro Ser Arg Thr
            740                 745                 750
Cys Val Val Tyr Ala Pro Ala Glu
        755                 760
 
           
             18 
             844 
             PRT 
             Oryza sativa 
           
            18
Val Glu Ala Glu Arg Gly Gly Cys Arg Gly Ile Arg Ser Gly Cys Gly
1               5                   10                  15
Ala Gly Glu Met Ala Ala Pro Ala Ser Ala Val Pro Gly Ser Ala Ala
            20                  25                  30
Gly Leu Arg Ala Gly Ala Val Arg Phe Pro Val Pro Ala Gly Ala Arg
        35                  40                  45
Ser Trp Arg Ala Ala Ala Glu Leu Pro Thr Ser Arg Ser Leu Leu Ser
    50                  55                  60
Gly Arg Arg Phe Pro Gly Ala Val Arg Val Gly Gly Ser Gly Gly Arg
65                  70                  75                  80
Val Ala Val Arg Ala Ala Gly Ala Ser Gly Glu Val Met Ile Pro Glu
                85                  90                  95
Gly Glu Ser Asp Gly Met Pro Val Ser Ala Gly Ser Asp Asp Leu Gln
            100                 105                 110
Leu Pro Ala Leu Asp Asp Glu Leu Ser Thr Glu Val Gly Ala Glu Val
        115                 120                 125
Glu Ile Glu Ser Ser Gly Ala Ser Asp Val Glu Gly Val Lys Arg Val
    130                 135                 140
Val Glu Glu Leu Ala Ala Glu Gln Lys Pro Arg Val Val Pro Pro Thr
145                 150                 155                 160
Gly Asp Gly Gln Lys Ile Phe Gln Met Asp Ser Met Leu Asn Gly Tyr
                165                 170                 175
Lys Tyr His Leu Glu Tyr Arg Tyr Ser Leu Tyr Arg Arg Leu Arg Ser
            180                 185                 190
Asp Ile Asp Gln Tyr Glu Gly Gly Leu Glu Thr Phe Ser Arg Gly Tyr
        195                 200                 205
Glu Lys Phe Gly Phe Asn His Ser Ala Glu Gly Val Thr Tyr Arg Glu
    210                 215                 220
Trp Ala Pro Gly Ala His Ser Ala Ala Leu Val Gly Asp Phe Asn Asn
225                 230                 235                 240
Trp Asn Pro Asn Ala Asp Arg Met Ser Lys Asn Glu Phe Gly Val Trp
                245                 250                 255
Glu Ile Phe Leu Pro Asn Asn Ala Asp Gly Ser Ser Pro Ile Pro His
            260                 265                 270
Gly Ser Arg Val Lys Val Arg Met Glu Thr Pro Ser Gly Ile Lys Asp
        275                 280                 285
Ser Ile Pro Ala Trp Ile Lys Tyr Ser Val Gln Ala Ala Gly Glu Ile
    290                 295                 300
Pro Tyr Asn Gly Ile Tyr Tyr Asp Pro Pro Glu Glu Glu Lys Tyr Ile
305                 310                 315                 320
Phe Lys His Pro Gln Pro Lys Arg Pro Lys Ser Leu Arg Ile Tyr Glu
                325                 330                 335
Thr His Val Gly Met Ser Ser Thr Glu Pro Lys Ile Asn Thr Tyr Ala
            340                 345                 350
Asn Phe Arg Asp Glu Val Leu Pro Arg Ile Lys Lys Leu Gly Tyr Asn
        355                 360                 365
Ala Val Gln Ile Met Ala Ile Gln Glu His Ala Tyr Tyr Gly Ser Phe
    370                 375                 380
Gly Tyr His Val Thr Asn Phe Phe Ala Pro Ser Ser Arg Phe Gly Thr
385                 390                 395                 400
Pro Glu Asp Leu Lys Ser Leu Ile Asp Lys Ala His Glu Leu Gly Leu
                405                 410                 415
Val Val Leu Met Asp Val Val His Ser His Ala Ser Asn Asn Thr Leu
            420                 425                 430
Asp Gly Leu Asn Gly Phe Asp Gly Thr Asp Thr His Tyr Phe His Ser
        435                 440                 445
Gly Ser Arg Gly His His Trp Met Trp Asp Ser Arg Leu Phe Asn Tyr
    450                 455                 460
Gly Asn Trp Glu Val Leu Arg Phe Leu Leu Ser Asn Ala Arg Trp Trp
465                 470                 475                 480
Leu Glu Glu Tyr Lys Phe Asp Gly Phe Arg Phe Asp Gly Val Thr Ser
                485                 490                 495
Met Met Tyr Thr His His Gly Leu Gln Val Ala Phe Thr Gly Asn Tyr
            500                 505                 510
Ser Glu Tyr Phe Gly Phe Ala Thr Asp Ala Asp Ala Val Val Tyr Leu
        515                 520                 525
Met Leu Val Asn Asp Leu Ile His Gly Leu Tyr Pro Glu Ala Ile Thr
    530                 535                 540
Ile Gly Glu Asp Val Ser Gly Met Pro Thr Phe Ala Leu Pro Val Gln
545                 550                 555                 560
Asp Gly Gly Val Gly Phe Asp Tyr Arg Leu His Met Ala Val Pro Asp
                565                 570                 575
Lys Trp Ile Glu Leu Leu Lys Gln Ser Asp Glu Ser Trp Lys Met Gly
            580                 585                 590
Asp Ile Val His Thr Leu Thr Asn Arg Arg Trp Ser Glu Lys Cys Val
        595                 600                 605
Thr Tyr Ala Glu Ser His Asp Gln Ala Leu Val Gly Asp Lys Thr Ile
    610                 615                 620
Ala Phe Trp Leu Met Asp Lys Asp Met Tyr Asp Phe Met Ala Leu Asp
625                 630                 635                 640
Arg Pro Ala Thr Pro Ser Ile Asp Arg Gly Ile Ala Leu His Lys Met
                645                 650                 655
Ile Arg Leu Ile Thr Met Gly Leu Gly Gly Glu Gly Tyr Leu Asn Phe
            660                 665                 670
Met Gly Asn Glu Phe Gly His Pro Glu Trp Ile Asp Phe Pro Arg Ala
        675                 680                 685
Pro Gln Val Leu Pro Asn Gly Lys Phe Ile Pro Gly Asn Asn Asn Ser
    690                 695                 700
Tyr Asp Lys Cys Arg Arg Arg Phe Asp Leu Gly Asp Ala Asp Tyr Leu
705                 710                 715                 720
Arg Tyr Arg Gly Met Leu Glu Phe Asp Arg Ala Met Gln Ser Leu Glu
                725                 730                 735
Glu Lys Tyr Gly Phe Met Thr Ser Asp His Gln Tyr Ile Ser Arg Lys
            740                 745                 750
His Glu Glu Asp Lys Met Ile Ile Phe Glu Lys Gly Asp Leu Val Phe
        755                 760                 765
Val Phe Asn Phe His Trp Ser Asn Ser Tyr Phe Asp Tyr Arg Val Gly
    770                 775                 780
Cys Leu Lys Pro Gly Lys Tyr Lys Val Val Leu Asp Ser Asp Ala Gly
785                 790                 795                 800
Leu Phe Gly Gly Phe Gly Arg Ile His His Thr Ala Glu His Phe Thr
                805                 810                 815
Ala Asp Cys Ser His Asp Asn Arg Pro Tyr Ser Phe Ser Val Tyr Ser
            820                 825                 830
Pro Ser Arg Thr Cys Val Val Tyr Ala Pro Ala Glu
        835                 840
 
           
             19 
             857 
             PRT 
             Pisum sativum 
           
            19
Lys Val Leu Ile Pro Glu Asp Gln Asp Asn Ser Val Ser Leu Ala Asp
1               5                   10                  15
Gln Leu Glu Asn Pro Asp Ile Thr Ser Glu Asp Ala Gln Asn Leu Glu
            20                  25                  30
Asp Leu Thr Met Lys Asp Gly Asn Lys Tyr Asn Ile Asp Glu Ser Thr
        35                  40                  45
Ser Ser Tyr Arg Glu Val Gly Asp Glu Lys Gly Ser Val Thr Ser Ser
    50                  55                  60
Ser Leu Val Asp Val Asn Thr Asp Thr Gln Ala Lys Lys Thr Ser Val
65                  70                  75                  80
His Ser Asp Lys Lys Val Lys Val Asp Lys Pro Lys Ile Ile Pro Pro
                85                  90                  95
Pro Gly Thr Gly Gln Lys Ile Tyr Glu Ile Asp Pro Leu Leu Gln Ala
            100                 105                 110
His Arg Gln His Leu Asp Phe Arg Tyr Gly Gln Tyr Lys Arg Ile Arg
        115                 120                 125
Glu Glu Ile Asp Lys Tyr Glu Gly Gly Leu Asp Ala Phe Ser Arg Gly
    130                 135                 140
Tyr Glu Lys Phe Gly Phe Thr Arg Ser Ala Thr Gly Ile Thr Tyr Arg
145                 150                 155                 160
Glu Trp Ala Pro Gly Ala Lys Ser Ala Ala Leu Val Gly Asp Phe Asn
                165                 170                 175
Asn Trp Asn Pro Asn Ala Asp Val Met Thr Lys Asp Ala Phe Gly Val
            180                 185                 190
Trp Glu Ile Phe Leu Pro Asn Asn Ala Asp Gly Ser Pro Pro Ile Pro
        195                 200                 205
His Gly Ser Arg Val Lys Ile His Met Asp Thr Pro Ser Gly Ile Lys
    210                 215                 220
Asp Ser Ile Pro Ala Trp Ile Lys Phe Ser Val Gln Ala Pro Gly Glu
225                 230                 235                 240
Ile Pro Tyr Asn Gly Ile Tyr Tyr Asp Pro Pro Glu Glu Glu Lys Tyr
                245                 250                 255
Val Phe Lys His Pro Gln Pro Lys Arg Pro Gln Ser Ile Arg Ile Tyr
            260                 265                 270
Glu Ser His Ile Gly Met Ser Ser Pro Glu Pro Lys Ile Asn Thr Tyr
        275                 280                 285
Ala Asn Phe Arg Asp Asp Val Leu Pro Arg Ile Lys Lys Leu Gly Tyr
    290                 295                 300
Asn Ala Val Gln Ile Met Ala Ile Gln Glu His Ser Tyr Tyr Ala Ser
305                 310                 315                 320
Phe Gly Tyr His Val Thr Asn Phe Phe Ala Pro Ser Ser Arg Phe Gly
                325                 330                 335
Thr Pro Glu Asp Leu Lys Ser Leu Ile Asp Arg Ala His Glu Leu Gly
            340                 345                 350
Leu Leu Val Leu Met Asp Ile Val His Ser His Ser Ser Asn Asn Thr
        355                 360                 365
Leu Asp Gly Leu Asn Met Phe Asp Gly Thr Asp Gly His Tyr Phe His
    370                 375                 380
Pro Gly Ser Arg Gly Tyr His Trp Met Trp Asp Ser Arg Leu Phe Asn
385                 390                 395                 400
Tyr Gly Ser Trp Glu Val Leu Arg Tyr Leu Leu Ser Asn Ala Arg Trp
                405                 410                 415
Trp Leu Asp Glu Tyr Lys Phe Asp Gly Phe Arg Phe Asp Gly Val Thr
            420                 425                 430
Ser Met Met Tyr Thr His His Gly Leu Gln Val Ser Phe Thr Gly Asn
        435                 440                 445
Tyr Ser Glu Tyr Phe Gly Leu Ala Thr Asp Val Glu Ala Val Val Tyr
    450                 455                 460
Met Met Leu Val Asn Asp Leu Ile His Gly Leu Phe Pro Glu Ala Val
465                 470                 475                 480
Ser Ile Gly Glu Asp Val Ser Gly Met Pro Thr Phe Cys Leu Pro Thr
                485                 490                 495
Gln Asp Gly Gly Ile Gly Phe Asn Tyr Arg Leu His Met Ala Val Ala
            500                 505                 510
Asp Lys Trp Ile Glu Leu Leu Lys Lys Gln Asp Glu Asp Trp Arg Met
        515                 520                 525
Gly Asp Ile Val His Thr Leu Thr Asn Arg Arg Trp Leu Glu Lys Cys
    530                 535                 540
Val Val Tyr Ala Glu Ser His Asp Gln Ala Leu Val Gly Asp Lys Thr
545                 550                 555                 560
Leu Ala Phe Trp Leu Met Asp Lys Asp Met Tyr Asp Phe Met Ala Leu
                565                 570                 575
Asp Arg Pro Ser Thr Pro Leu Ile Asp Arg Gly Ile Ala Leu His Lys
            580                 585                 590
Met Ile Arg Leu Ile Thr Met Gly Leu Gly Gly Glu Gly Tyr Leu Asn
        595                 600                 605
Phe Met Gly Asn Glu Phe Gly His Pro Glu Trp Ile Asp Phe Pro Arg
    610                 615                 620
Gly Glu Gln His Leu Pro Asn Gly Lys Ile Val Pro Gly Asn Asn Asn
625                 630                 635                 640
Ser Tyr Asp Lys Cys Arg Arg Arg Phe Asp Leu Gly Asp Ala Asp Tyr
                645                 650                 655
Leu Arg Tyr His Gly Met Gln Glu Phe Asp Arg Ala Met Gln His Leu
            660                 665                 670
Glu Glu Arg Tyr Gly Phe Met Thr Ser Glu His Gln Tyr Ile Ser Arg
        675                 680                 685
Lys Asn Glu Gly Asp Arg Val Ile Ile Phe Glu Arg Asp Asn Leu Val
    690                 695                 700
Phe Val Phe Asn Phe His Trp Thr Asn Ser Tyr Ser Asp Tyr Lys Val
705                 710                 715                 720
Gly Cys Leu Lys Pro Gly Lys Tyr Lys Ile Val Leu Asp Ser Asp Asp
                725                 730                 735
Thr Leu Phe Gly Gly Phe Asn Arg Leu Asn His Thr Ala Glu Tyr Phe
            740                 745                 750
Thr Ser Glu Gly Trp Tyr Asp Asp Arg Pro Arg Ser Phe Leu Val Tyr
        755                 760                 765
Ala Pro Ser Arg Thr Ala Val Val Tyr Ala Leu Ala Asp Gly Val Glu
    770                 775                 780
Ser Glu Pro Ile Glu Leu Ser Asp Gly Val Glu Ser Glu Pro Ile Glu
785                 790                 795                 800
Leu Ser Val Gly Val Glu Ser Glu Pro Ile Glu Leu Ser Val Glu Glu
                805                 810                 815
Ala Glu Ser Glu Pro Ile Glu Arg Ser Val Glu Glu Val Glu Ser Glu
            820                 825                 830
Thr Thr Gln Gln Ser Val Glu Val Glu Ser Glu Thr Thr Gln Gln Ser
        835                 840                 845
Val Glu Val Glu Ser Glu Thr Thr Gln
    850                 855
 
           
             20 
             779 
             PRT 
             Solanum tuberosum 
           
            20
Thr Met Ala Pro Leu Glu Glu Asp Val Lys Thr Glu Asn Ile Gly Leu
1               5                   10                  15
Leu Asn Leu Asp Pro Thr Leu Glu Pro Tyr Leu Asp His Phe Arg His
            20                  25                  30
Arg Met Lys Arg Tyr Val Asp Gln Lys Met Leu Ile Glu Lys Tyr Glu
        35                  40                  45
Gly Pro Leu Glu Glu Phe Ala Gln Gly Tyr Leu Lys Phe Gly Phe Asn
    50                  55                  60
Arg Glu Asp Gly Cys Ile Val Tyr Arg Glu Trp Ala Pro Ala Ala Gln
65                  70                  75                  80
Glu Asp Glu Val Ile Gly Asp Phe Asn Gly Trp Asn Gly Ser Asn His
                85                  90                  95
Met Met Glu Lys Asp Gln Phe Gly Val Trp Ser Ile Arg Ile Pro Asp
            100                 105                 110
Val Asp Ser Lys Pro Val Ile Pro His Asn Ser Arg Val Lys Phe Arg
        115                 120                 125
Phe Lys His Gly Asn Gly Val Trp Val Asp Arg Ile Pro Ala Trp Ile
    130                 135                 140
Lys Tyr Ala Thr Ala Asp Ala Thr Lys Phe Ala Ala Pro Tyr Asp Gly
145                 150                 155                 160
Val Tyr Trp Asp Pro Pro Pro Ser Glu Arg Tyr His Phe Lys Tyr Pro
                165                 170                 175
Arg Pro Pro Lys Pro Arg Ala Pro Arg Ile Tyr Glu Ala His Val Gly
            180                 185                 190
Met Ser Ser Ser Glu Pro Arg Val Asn Ser Tyr Arg Glu Phe Ala Asp
        195                 200                 205
Asp Val Leu Pro Arg Ile Lys Ala Asn Asn Tyr Asn Thr Val Gln Leu
    210                 215                 220
Met Ala Ile Met Glu His Ser Tyr Tyr Gly Ser Phe Gly Tyr His Val
225                 230                 235                 240
Thr Asn Phe Phe Ala Val Ser Ser Arg Tyr Gly Asn Pro Glu Asp Leu
                245                 250                 255
Lys Tyr Leu Ile Asp Lys Ala His Ser Leu Gly Leu Gln Val Leu Val
            260                 265                 270
Asp Val Val His Ser His Ala Ser Asn Asn Val Thr Asp Gly Leu Asn
        275                 280                 285
Gly Phe Asp Ile Gly Gln Gly Ser Gln Glu Ser Tyr Phe His Ala Gly
    290                 295                 300
Glu Arg Gly Tyr His Lys Leu Trp Asp Ser Arg Leu Phe Asn Tyr Ala
305                 310                 315                 320
Asn Trp Glu Val Leu Arg Phe Leu Leu Ser Asn Leu Arg Trp Trp Leu
                325                 330                 335
Glu Glu Tyr Asn Phe Asp Gly Phe Arg Phe Asp Gly Ile Thr Ser Met
            340                 345                 350
Leu Tyr Val His His Gly Ile Asn Met Gly Phe Thr Gly Asn Tyr Asn
        355                 360                 365
Glu Tyr Phe Ser Glu Ala Thr Asp Val Asp Ala Val Val Tyr Leu Met
    370                 375                 380
Leu Ala Asn Asn Leu Ile His Lys Ile Phe Pro Asp Ala Thr Val Ile
385                 390                 395                 400
Ala Glu Asp Val Ser Gly Met Pro Gly Leu Gly Arg Pro Val Ser Glu
                405                 410                 415
Gly Gly Ile Gly Phe Asp Tyr Arg Leu Ala Met Ala Ile Pro Asp Lys
            420                 425                 430
Trp Ile Asp Tyr Leu Lys Asn Lys Asn Asp Glu Asp Trp Ser Met Lys
        435                 440                 445
Glu Val Thr Ser Ser Leu Thr Asn Arg Arg Tyr Thr Glu Lys Cys Ile
    450                 455                 460
Ala Tyr Ala Glu Ser His Asp Gln Ser Ile Val Gly Asp Lys Thr Ile
465                 470                 475                 480
Ala Phe Leu Leu Met Asp Lys Glu Met Tyr Ser Gly Met Ser Cys Leu
                485                 490                 495
Thr Asp Ala Ser Pro Val Val Asp Arg Gly Ile Ala Leu His Lys Met
            500                 505                 510
Ile His Phe Phe Thr Met Ala Leu Gly Gly Glu Gly Tyr Leu Asn Phe
        515                 520                 525
Met Gly Asn Glu Phe Gly His Pro Glu Trp Ile Asp Phe Pro Arg Glu
    530                 535                 540
Gly Asn Asn Trp Ser Tyr Asp Lys Cys Arg Arg Gln Trp Asn Leu Ala
545                 550                 555                 560
Asp Ser Glu His Leu Arg Tyr Lys Phe Met Asn Ala Phe Asp Arg Ala
                565                 570                 575
Met Asn Ser Leu Asp Glu Lys Phe Ser Phe Leu Ala Ser Gly Lys Gln
            580                 585                 590
Ile Val Ser Ser Met Asp Asp Asp Asn Lys Val Val Val Phe Glu Arg
        595                 600                 605
Gly Asp Leu Val Phe Val Phe Asn Phe His Pro Lys Asn Thr Tyr Glu
    610                 615                 620
Gly Tyr Lys Val Gly Cys Asp Leu Pro Gly Lys Tyr Arg Val Ala Leu
625                 630                 635                 640
Asp Ser Asp Ala Trp Glu Phe Gly Gly His Gly Arg Thr Gly His Asp
                645                 650                 655
Val Asp His Phe Thr Ser Pro Glu Gly Ile Pro Gly Val Pro Glu Thr
            660                 665                 670
Asn Phe Asn Gly Arg Gln Ile Pro Ser Lys Cys Cys Leu Leu Arg Glu
        675                 680                 685
His Val Trp Leu Ile Thr Glu Leu Met Asn Ala Cys Gln Lys Leu Lys
    690                 695                 700
Ile Thr Arg Gln Thr Phe Val Val Ser Tyr Tyr Gln Gln Pro Ile Ser
705                 710                 715                 720
Arg Arg Val Thr Arg Asn Leu Lys Ile Arg Tyr Leu Gln Ile Ser Val
                725                 730                 735
Thr Leu Thr Asn Ala Cys Gln Lys Leu Lys Phe Thr Arg Gln Thr Phe
            740                 745                 750
Leu Val Ser Tyr Tyr Gln Gln Pro Ile Leu Arg Arg Val Thr Arg Lys
        755                 760                 765
Leu Lys Asp Ser Leu Ser Thr Asn Ile Ser Thr
    770                 775
 
           
             21 
             762 
             PRT 
             Triticum aestivum 
           
            21
Thr Met Ala Thr Ala Glu Asp Gly Val Gly Asp Leu Pro Ile Tyr Asp
1               5                   10                  15
Leu Asp Pro Lys Phe Ala Gly Phe Lys Glu His Phe Ser Tyr Arg Met
            20                  25                  30
Lys Lys Tyr Leu Asp Gln Lys His Ser Ile Glu Lys His Glu Gly Gly
        35                  40                  45
Leu Glu Glu Phe Ser Lys Gly Tyr Leu Lys Phe Gly Ile Asn Thr Glu
    50                  55                  60
Asn Asp Ala Thr Val Tyr Arg Glu Trp Ala Pro Ala Ala Met Asp Ala
65                  70                  75                  80
Gln Leu Ile Gly Asp Phe Asn Asn Trp Asn Gly Ser Gly His Arg Met
                85                  90                  95
Thr Lys Asp Asn Tyr Gly Val Trp Ser Ile Arg Ile Ser His Val Asn
            100                 105                 110
Gly Lys Pro Ala Ile Pro His Asn Ser Lys Val Lys Phe Arg Phe His
        115                 120                 125
Arg Gly Asp Gly Leu Trp Val Asp Arg Val Pro Ala Trp Ile Arg Tyr
    130                 135                 140
Ala Thr Phe Asp Ala Ser Lys Phe Gly Ala Pro Tyr Asp Gly Val His
145                 150                 155                 160
Trp Asp Pro Pro Ser Gly Glu Arg Tyr Val Phe Lys His Pro Arg Pro
                165                 170                 175
Arg Lys Pro Asp Ala Pro Arg Ile Tyr Glu Ala His Val Gly Met Ser
            180                 185                 190
Gly Glu Lys Pro Glu Val Ser Thr Tyr Arg Glu Phe Ala Asp Asn Val
        195                 200                 205
Leu Pro Arg Ile Lys Ala Asn Asn Tyr Asn Thr Val Gln Leu Met Ala
    210                 215                 220
Ile Met Glu His Ser Tyr Tyr Ala Ser Phe Gly Tyr His Val Thr Asn
225                 230                 235                 240
Phe Phe Ala Val Ser Ser Arg Ser Gly Thr Pro Glu Asp Leu Lys Tyr
                245                 250                 255
Leu Val Asp Lys Ala His Ser Leu Gly Leu Arg Val Leu Met Asp Val
            260                 265                 270
Val His Ser His Ala Ser Ser Asn Lys Thr Asp Gly Leu Asn Gly Tyr
        275                 280                 285
Asp Val Gly Gln Asn Thr Gln Glu Ser Tyr Phe His Thr Gly Glu Arg
    290                 295                 300
Gly Tyr His Lys Leu Trp Asp Ser Arg Leu Phe Asn Tyr Ala Asn Trp
305                 310                 315                 320
Glu Val Leu Arg Phe Leu Leu Ser Asn Leu Arg Tyr Trp Met Asp Glu
                325                 330                 335
Phe Met Phe Asp Gly Phe Arg Phe Asp Gly Val Thr Ser Met Leu Tyr
            340                 345                 350
Asn His His Gly Ile Asn Met Ser Phe Ala Gly Ser Tyr Lys Glu Tyr
        355                 360                 365
Phe Gly Leu Asp Thr Asp Val Asp Ala Val Val Tyr Leu Met Leu Ala
    370                 375                 380
Asn His Leu Met His Lys Leu Leu Pro Glu Ala Thr Val Val Ala Glu
385                 390                 395                 400
Asp Val Ser Gly Met Pro Val Leu Cys Arg Ser Val Asp Glu Gly Gly
                405                 410                 415
Val Gly Phe Asp Tyr Arg Leu Ala Met Ala Ile Pro Asp Arg Trp Ile
            420                 425                 430
Asp Tyr Leu Lys Asn Lys Asp Asp Leu Glu Trp Ser Met Ser Gly Ile
        435                 440                 445
Ala His Thr Leu Thr Asn Arg Arg Tyr Thr Glu Lys Cys Ile Ala Tyr
    450                 455                 460
Ala Glu Ser His Asp Gln Ser Ile Val Gly Asp Lys Thr Met Ala Phe
465                 470                 475                 480
Leu Leu Met Asp Lys Glu Met Tyr Thr Gly Met Ser Asp Leu Gln Pro
                485                 490                 495
Ala Ser Pro Thr Ile Asp Arg Gly Ile Ala Leu Gln Lys Met Ile His
            500                 505                 510
Phe Ile Thr Met Ala Leu Gly Gly Asp Gly Tyr Leu Asn Phe Met Gly
        515                 520                 525
Asn Glu Phe Gly His Pro Glu Trp Ile Asp Phe Pro Arg Glu Gly Asn
    530                 535                 540
Asn Trp Ser Tyr Asp Lys Cys Arg Arg Gln Trp Ser Leu Ala Asp Ile
545                 550                 555                 560
Asp His Leu Arg Tyr Lys Tyr Met Asn Ala Phe Asp Gln Ala Met Asn
                565                 570                 575
Ala Leu Asp Asp Lys Phe Ser Phe Leu Ser Ser Ser Lys Gln Ile Val
            580                 585                 590
Ser Asp Met Asn Glu Glu Lys Lys Ile Ile Val Phe Glu Arg Gly Asp
        595                 600                 605
Leu Val Phe Val Phe Asn Phe His Pro Ser Lys Thr Tyr Asp Gly Tyr
    610                 615                 620
Lys Val Gly Cys Asp Leu Pro Gly Lys Tyr Lys Val Ala Leu Asp Ser
625                 630                 635                 640
Asp Ala Leu Met Phe Gly Gly His Gly Arg Val Ala His Asp Asn Asp
                645                 650                 655
His Phe Thr Ser Pro Glu Gly Val Pro Gly Val Pro Glu Thr Asn Phe
            660                 665                 670
Asn Asn Arg Pro Asn Ser Phe Lys Ile Leu Ser Pro Ser Arg Thr Cys
        675                 680                 685
Val Ala Tyr Tyr Arg Val Glu Glu Lys Ala Glu Lys Pro Lys Asp Glu
    690                 695                 700
Gly Ala Ala Ser Trp Gly Lys Thr Ala Leu Gly Tyr Ile Asp Val Glu
705                 710                 715                 720
Ala Thr Gly Val Lys Asp Ala Ala Asp Gly Glu Ala Thr Ser Gly Ser
                725                 730                 735
Glu Lys Ala Ser Thr Gly Gly Asp Ser Ser Lys Lys Gly Ile Asn Phe
            740                 745                 750
Val Phe Leu Ser Pro Asp Lys Asp Asn Lys
        755                 760
 
           
             22 
             703 
             PRT 
             Triticum aestivum 
           
            22
Ser Pro Pro Thr Leu Thr Ser Pro Pro Pro Ser Ala Val Pro Ser Thr
1               5                   10                  15
Thr Met Leu Cys Leu Ser Ser Ser Leu Leu Pro Arg Pro Ser Ala Ala
            20                  25                  30
Ala Asp Arg Pro Leu Pro Gly Ile Ile Ala Gly Gly Gly Gly Gly Lys
        35                  40                  45
Arg Leu Ser Val Val Pro Ser Val Pro Phe Leu Leu Arg Trp Leu Trp
    50                  55                  60
Pro Arg Lys Ala Lys Ser Lys Ser Phe Val Ser Val Thr Ala Arg Gly
65                  70                  75                  80
Asn Lys Ile Ala Ala Thr Thr Gly Tyr Gly Ser Asp His Leu Pro Ile
                85                  90                  95
Tyr Asp Leu Asp Leu Lys Leu Ala Glu Phe Lys Asp His Phe Asp Tyr
            100                 105                 110
Thr Arg Asn Arg Tyr Ile Glu Gln Lys His Leu Ile Glu Lys His Glu
        115                 120                 125
Gly Ser Leu Glu Glu Phe Ser Lys Gly Tyr Leu Lys Phe Gly Ile Asn
    130                 135                 140
Thr Glu His Gly Ala Ser Val Tyr Arg Glu Trp Ala Pro Ala Ala Glu
145                 150                 155                 160
Glu Ala Gln Leu Val Gly Asp Phe Asn Asn Trp Asn Gly Ser Gly His
                165                 170                 175
Lys Met Ala Lys Asp Asn Phe Gly Val Trp Ser Ile Arg Ile Ser His
            180                 185                 190
Val Asn Gly Lys Pro Ala Ile Pro His Asn Ser Lys Val Lys Phe Arg
        195                 200                 205
Phe Arg His His Gly Val Trp Val Glu Gln Ile Pro Ala Trp Ile Arg
    210                 215                 220
Tyr Ala Thr Val Thr Ala Ser Glu Ser Gly Ala Pro Tyr Asp Gly Leu
225                 230                 235                 240
His Trp Asp Pro Pro Ser Ser Glu Arg Tyr Val Phe Asn His Pro Arg
                245                 250                 255
Pro Pro Lys Pro Asp Val Pro Arg Ile Tyr Glu Ala His Val Gly Val
            260                 265                 270
Ser Gly Gly Lys Leu Glu Ala Gly Thr Tyr Arg Glu Phe Pro Asp Asn
        275                 280                 285
Val Leu Pro Cys Leu Arg Ala Thr Asn Tyr Asn Thr Val Gln Leu Met
    290                 295                 300
Gly Ile Met Glu His Ser Asp Ser Ala Ser Phe Gly Tyr His Val Thr
305                 310                 315                 320
Asn Phe Phe Ala Val Ser Ser Arg Ser Gly Thr Pro Glu Asp Leu Lys
                325                 330                 335
Tyr Leu Ile Asp Lys Ala His Ser Leu Gly Leu Arg Val Leu Met Asp
            340                 345                 350
Val Val His Ser His Ala Ser Asn Asn Val Ile Asp Gly Leu Asn Gly
        355                 360                 365
Tyr Asp Val Gly Gln Ser Ala His Glu Ser Tyr Phe Tyr Thr Gly Asp
    370                 375                 380
Lys Gly Tyr Asn Lys Met Trp Asn Gly Arg Met Phe Asn Tyr Ala Asn
385                 390                 395                 400
Trp Glu Val Leu Arg Phe Leu Leu Ser Asn Leu Arg Tyr Trp Met Asp
                405                 410                 415
Glu Phe Met Phe Asp Gly Phe Arg Phe Val Gly Val Thr Ser Met Leu
            420                 425                 430
Tyr Asn His Asn Gly Ile Asn Met Ser Phe Asn Gly Asn Tyr Lys Asp
        435                 440                 445
Tyr Ile Gly Leu Asp Thr Asn Val Asp Ala Phe Val Tyr Met Met Leu
    450                 455                 460
Ala Asn His Leu Met His Lys Leu Phe Pro Glu Ala Ile Val Val Ala
465                 470                 475                 480
Val Asp Val Ser Gly Met Pro Val Leu Cys Trp Pro Val Asp Glu Gly
                485                 490                 495
Gly Leu Gly Phe Asp Tyr Arg Gln Ala Met Thr Ile Pro Asp Arg Trp
            500                 505                 510
Ile Asp Tyr Leu Glu Asn Lys Gly Asp Gln Gln Trp Ser Met Ser Ser
        515                 520                 525
Val Ile Ser Gln Thr Leu Thr Asn Arg Arg Tyr Pro Glu Lys Phe Ile
    530                 535                 540
Ala Tyr Ala Glu Arg Gln Asn His Ser Ile Ile Gly Ser Lys Thr Met
545                 550                 555                 560
Ala Phe Leu Leu Met Glu Trp Glu Thr Tyr Ser Gly Met Ser Ala Met
                565                 570                 575
Asp Pro Asp Ser Pro Thr Ile Asp Arg Ala Ile Ala Leu Gln Lys Met
            580                 585                 590
Ile His Phe Ile Thr Met Ala Phe Gly Gly Asp Ser Tyr Leu Lys Phe
        595                 600                 605
Met Gly Asn Glu Tyr Met Asn Ala Phe Val Gln Ala Val Asp Thr Pro
    610                 615                 620
Ser Asp Lys Cys Ser Phe Leu Ser Ser Ser Asn Gln Thr Ala Ser His
625                 630                 635                 640
Met Asn Glu Glu Glu Lys Gly Ser Ala Leu Thr Lys Gly Tyr Thr His
                645                 650                 655
Leu Arg Ser Gly Cys Phe Asp Pro Ser Leu Pro Ser Thr Ser Ser Cys
            660                 665                 670
Ala Phe Leu Gly Pro Ser Asn Gln Ser Pro Phe Ser Lys Pro Phe Ile
        675                 680                 685
Gly Phe Pro Gly Cys Ile Phe Cys Cys Gly Leu Phe Lys Gly Glu
    690                 695                 700
 
           
             23 
             752 
             PRT 
             Zea mays 
           
            23
Thr Met Ala Thr Ala Lys Gly Asp Val Asp His Leu Pro Ile Tyr Asp
1               5                   10                  15
Leu Asp Pro Lys Leu Glu Ile Phe Lys Asp His Phe Arg Tyr Arg Met
            20                  25                  30
Lys Arg Phe Leu Glu Gln Lys Gly Ser Ile Glu Glu Asn Glu Gly Ser
        35                  40                  45
Leu Glu Ser Phe Ser Lys Gly Tyr Leu Lys Phe Gly Ile Asn Thr Asn
    50                  55                  60
Glu Asp Gly Thr Val Tyr Arg Glu Trp Ala Pro Ala Ala Gln Glu Ala
65                  70                  75                  80
Glu Leu Ile Gly Asp Phe Asn Asp Trp Asn Gly Ala Asn His Lys Met
                85                  90                  95
Glu Lys Asp Lys Phe Gly Val Trp Ser Ile Lys Ile Asp His Val Lys
            100                 105                 110
Gly Lys Pro Ala Ile Pro His Asn Ser Lys Val Lys Phe Arg Phe Leu
        115                 120                 125
His Gly Gly Val Trp Val Asp Arg Ile Pro Ala Leu Ile Arg Tyr Ala
    130                 135                 140
Thr Val Asp Ala Ser Lys Phe Gly Ala Pro Tyr Asp Gly Val His Trp
145                 150                 155                 160
Asp Pro Pro Ala Ser Glu Arg Tyr Thr Phe Lys His Pro Arg Pro Ser
                165                 170                 175
Lys Pro Ala Ala Pro Arg Ile Tyr Glu Ala His Val Gly Met Ser Gly
            180                 185                 190
Glu Lys Pro Ala Val Ser Thr Tyr Arg Glu Phe Ala Asp Asn Val Leu
        195                 200                 205
Pro Arg Ile Arg Ala Asn Asn Tyr Asn Thr Val Gln Leu Met Ala Val
    210                 215                 220
Met Glu His Ser Tyr Tyr Ala Ser Phe Gly Tyr His Val Thr Asn Phe
225                 230                 235                 240
Phe Ala Val Ser Ser Arg Ser Gly Thr Pro Glu Asp Leu Lys Tyr Leu
                245                 250                 255
Val Asp Lys Ala His Ser Leu Gly Leu Arg Val Leu Met Asp Val Val
            260                 265                 270
His Ser His Ala Ser Asn Asn Val Thr Asp Gly Leu Asn Gly Tyr Asp
        275                 280                 285
Val Gly Gln Ser Thr Gln Glu Ser Tyr Phe His Ala Gly Asp Arg Gly
    290                 295                 300
Tyr His Lys Leu Trp Asp Ser Arg Leu Phe Asn Tyr Ala Asn Trp Glu
305                 310                 315                 320
Val Leu Arg Phe Leu Leu Ser Asn Leu Arg Tyr Trp Leu Asp Glu Phe
                325                 330                 335
Met Phe Asp Gly Phe Arg Phe Asp Gly Val Thr Ser Met Leu Tyr His
            340                 345                 350
His His Gly Ile Asn Val Gly Phe Thr Gly Asn Tyr Gln Glu Tyr Phe
        355                 360                 365
Ser Leu Asp Thr Ala Val Asp Ala Val Val Tyr Met Met Leu Ala Asn
    370                 375                 380
His Leu Met His Lys Leu Leu Pro Glu Ala Thr Val Val Ala Glu Asp
385                 390                 395                 400
Val Ser Gly Met Pro Val Leu Cys Arg Pro Val Asp Glu Gly Gly Val
                405                 410                 415
Gly Phe Asp Tyr Arg Leu Ala Met Ala Ile Pro Asp Arg Trp Ile Asp
            420                 425                 430
Tyr Leu Lys Asn Lys Asp Asp Ser Glu Trp Ser Met Gly Glu Ile Ala
        435                 440                 445
His Thr Leu Thr Asn Arg Arg Tyr Thr Glu Lys Cys Ile Ala Tyr Ala
    450                 455                 460
Glu Ser His Asp Gln Ser Ile Val Gly Asp Lys Thr Ile Ala Phe Leu
465                 470                 475                 480
Leu Met Asp Lys Glu Met Tyr Thr Gly Met Ser Asp Leu Gln Pro Ala
                485                 490                 495
Ser Pro Thr Ile Asp Arg Gly Ile Ala Leu Gln Lys Met Ile His Phe
            500                 505                 510
Ile Thr Met Ala Leu Gly Gly Asp Gly Tyr Leu Asn Phe Met Gly Asn
        515                 520                 525
Glu Phe Gly His Pro Glu Trp Ile Asp Phe Pro Arg Glu Gly Asn Asn
    530                 535                 540
Trp Ser Tyr Asp Lys Cys Arg Arg Gln Trp Ser Leu Val Asp Thr Asp
545                 550                 555                 560
His Leu Arg Tyr Lys Tyr Met Asn Ala Phe Asp Gln Ala Met Asn Ala
                565                 570                 575
Leu Asp Glu Arg Phe Ser Phe Leu Ser Ser Ser Lys Gln Ile Val Ser
            580                 585                 590
Asp Met Asn Asp Glu Glu Lys Val Ile Val Phe Glu Arg Gly Asp Leu
        595                 600                 605
Val Phe Val Phe Asn Phe His Pro Lys Lys Thr Tyr Glu Gly Tyr Lys
    610                 615                 620
Val Gly Cys Asp Leu Pro Gly Lys Tyr Arg Val Ala Leu Asp Ser Asp
625                 630                 635                 640
Ala Leu Val Phe Gly Gly His Gly Arg Val Gly His Asp Val Asp His
                645                 650                 655
Phe Thr Ser Pro Glu Gly Val Pro Gly Val Pro Glu Thr Asn Phe Asn
            660                 665                 670
Asn Arg Pro Asn Ser Phe Lys Val Leu Ser Pro Pro Arg Thr Cys Val
        675                 680                 685
Ala Tyr Tyr Arg Val Asp Glu Ala Gly Ala Gly Arg Arg Leu His Ala
    690                 695                 700
Lys Ala Glu Thr Gly Lys Thr Ser Pro Ala Glu Ser Ile Asp Val Lys
705                 710                 715                 720
Ala Ser Arg Ala Ser Ser Lys Glu Asp Lys Glu Ala Thr Ala Gly Gly
                725                 730                 735
Lys Lys Gly Trp Lys Phe Ala Arg Gln Pro Ser Asp Gln Asp Thr Lys
            740                 745                 750
 
           
             24 
             756 
             PRT 
             Oryza sativa 
           
            24
Thr Met Val Thr Val Val Glu Glu Val Asp His Leu Pro Ile Tyr Asp
1               5                   10                  15
Leu Asp Pro Lys Leu Glu Glu Phe Lys Asp His Phe Asn Tyr Arg Ile
            20                  25                  30
Lys Arg Tyr Leu Asp Gln Lys Cys Leu Ile Glu Lys His Glu Gly Gly
        35                  40                  45
Leu Glu Glu Phe Ser Lys Gly Tyr Leu Lys Phe Gly Ile Asn Thr Val
    50                  55                  60
Asp Gly Ala Thr Ile Tyr Arg Glu Trp Ala Pro Ala Ala Gln Glu Ala
65                  70                  75                  80
Gln Leu Ile Gly Glu Phe Asn Asn Trp Asn Gly Ala Lys His Lys Met
                85                  90                  95
Glu Lys Asp Lys Phe Gly Ile Trp Ser Ile Lys Ile Ser His Val Asn
            100                 105                 110
Gly Lys Pro Ala Ile Pro His Asn Ser Lys Val Lys Phe Arg Phe Arg
        115                 120                 125
His Gly Gly Gly Ala Trp Val Asp Arg Ile Pro Ala Trp Ile Arg Tyr
    130                 135                 140
Ala Thr Phe Asp Ala Ser Lys Phe Gly Ala Pro Tyr Asp Gly Val His
145                 150                 155                 160
Trp Asp Pro Pro Ala Cys Glu Arg Tyr Val Phe Lys His Pro Arg Pro
                165                 170                 175
Pro Lys Pro Asp Ala Pro Arg Ile Tyr Glu Ala His Val Gly Met Ser
            180                 185                 190
Gly Glu Glu Pro Glu Val Ser Thr Tyr Arg Glu Phe Ala Asp Asn Val
        195                 200                 205
Leu Pro Arg Ile Arg Ala Asn Asn Tyr Asn Thr Val Gln Leu Met Ala
    210                 215                 220
Ile Met Glu His Ser Tyr Tyr Ala Ser Phe Gly Tyr His Val Thr Asn
225                 230                 235                 240
Phe Phe Ala Val Ser Ser Arg Ser Gly Thr Pro Glu Asp Leu Lys Tyr
                245                 250                 255
Leu Val Asp Lys Ala His Ser Leu Gly Leu Arg Val Leu Met Asp Val
            260                 265                 270
Val His Ser His Ala Ser Asn Asn Val Thr Asp Gly Leu Asn Gly Tyr
        275                 280                 285
Asp Val Gly Gln Asn Thr His Glu Ser Tyr Phe His Thr Gly Asp Arg
    290                 295                 300
Gly Tyr His Lys Leu Trp Asp Ser Arg Leu Phe Asn Tyr Ala Asn Trp
305                 310                 315                 320
Glu Val Leu Arg Phe Leu Leu Ser Asn Leu Arg Tyr Trp Met Asp Glu
                325                 330                 335
Phe Met Phe Asp Gly Phe Arg Phe Asp Gly Val Thr Ser Met Leu Tyr
            340                 345                 350
His His His Gly Ile Asn Lys Gly Phe Thr Gly Asn Tyr Lys Glu Tyr
        355                 360                 365
Phe Ser Leu Asp Thr Asp Val Asp Ala Ile Val Tyr Met Met Leu Ala
    370                 375                 380
Asn His Leu Met His Lys Leu Leu Pro Glu Ala Thr Ile Val Ala Glu
385                 390                 395                 400
Asp Val Ser Gly Met Pro Val Leu Cys Arg Pro Val Asp Glu Gly Gly
                405                 410                 415
Val Gly Phe Asp Phe Arg Leu Ala Met Ala Ile Pro Asp Arg Trp Ile
            420                 425                 430
Asp Tyr Leu Lys Asn Lys Glu Asp Arg Lys Trp Ser Met Ser Glu Ile
        435                 440                 445
Val Gln Thr Leu Thr Asn Arg Arg Tyr Thr Glu Lys Cys Ile Ala Tyr
    450                 455                 460
Ala Glu Ser His Asp Gln Ser Ile Val Gly Asp Lys Thr Ile Ala Phe
465                 470                 475                 480
Leu Leu Met Asp Lys Glu Met Tyr Thr Gly Met Ser Asp Leu Gln Pro
                485                 490                 495
Ala Ser Pro Thr Ile Asn Arg Gly Ile Ala Leu Gln Lys Met Ile His
            500                 505                 510
Phe Ile Thr Met Ala Leu Gly Gly Asp Gly Tyr Leu Asn Phe Met Gly
        515                 520                 525
Asn Glu Phe Gly His Pro Glu Trp Ile Asp Phe Pro Arg Glu Gly Asn
    530                 535                 540
Asn Trp Ser Tyr Asp Lys Cys Arg Arg Gln Trp Ser Leu Val Asp Thr
545                 550                 555                 560
Asp His Leu Arg Tyr Lys Tyr Met Asn Ala Phe Asp Gln Ala Met Asn
                565                 570                 575
Ala Leu Glu Glu Glu Phe Ser Phe Leu Ser Ser Ser Lys Gln Ile Val
            580                 585                 590
Ser Asp Met Asn Glu Lys Asp Lys Val Ile Val Phe Glu Arg Gly Asp
        595                 600                 605
Leu Val Phe Val Phe Asn Phe His Pro Asn Lys Thr Tyr Lys Gly Tyr
    610                 615                 620
Lys Val Gly Cys Asp Leu Pro Gly Lys Tyr Arg Val Ala Leu Asp Ser
625                 630                 635                 640
Asp Ala Leu Val Phe Gly Gly His Gly Arg Val Gly His Asp Val Asp
                645                 650                 655
His Phe Thr Ser Pro Glu Gly Met Pro Gly Val Pro Glu Thr Asn Phe
            660                 665                 670
Asn Asn Arg Pro Asn Ser Phe Lys Val Leu Ser Pro Pro Arg Thr Cys
        675                 680                 685
Val Ala Tyr Tyr Arg Val Asp Glu Asp Arg Glu Glu Leu Arg Arg Gly
    690                 695                 700
Gly Ala Val Ala Ser Gly Lys Ile Val Thr Glu Tyr Ile Asp Val Glu
705                 710                 715                 720
Ala Thr Ser Gly Glu Thr Ile Ser Gly Gly Trp Lys Gly Ser Glu Lys
                725                 730                 735
Asp Asp Cys Gly Lys Lys Gly Met Lys Phe Val Phe Arg Ser Ser Asp
            740                 745                 750
Glu Asp Cys Lys
        755
 
           
             25 
             762 
             PRT 
             Pisum sativum 
           
            25
Thr Met Pro Ser Val Glu Glu Asp Phe Glu Asn Ile Gly Ile Leu Asn
1               5                   10                  15
Val Asp Ser Ser Leu Glu Pro Phe Lys Asp His Phe Lys Tyr Arg Leu
            20                  25                  30
Lys Arg Tyr Leu His Gln Lys Lys Leu Ile Glu Glu Tyr Glu Gly Gly
        35                  40                  45
Leu Gln Glu Phe Ala Lys Gly Tyr Leu Lys Phe Gly Phe Asn Arg Glu
    50                  55                  60
Glu Asp Gly Ile Ser Tyr Arg Glu Trp Ala Pro Ala Ala Gln Glu Ala
65                  70                  75                  80
Gln Ile Ile Gly Asp Phe Asn Gly Trp Asn Gly Ser Asn Leu His Met
                85                  90                  95
Glu Lys Asp Gln Phe Gly Val Trp Ser Ile Gln Ile Pro Asp Ala Asp
            100                 105                 110
Gly Asn Pro Ala Ile Pro His Asn Ser Arg Val Lys Phe Arg Phe Lys
        115                 120                 125
His Ser Asp Gly Val Trp Val Asp Arg Ile Pro Ala Trp Ile Lys Tyr
    130                 135                 140
Ala Thr Val Asp Pro Thr Arg Phe Ala Ala Pro Tyr Asp Gly Val Tyr
145                 150                 155                 160
Trp Asp Pro Pro Leu Ser Glu Arg Tyr Gln Phe Lys His Pro Arg Pro
                165                 170                 175
Pro Lys Pro Lys Ala Pro Arg Ile Tyr Glu Ala His Val Gly Met Ser
            180                 185                 190
Ser Ser Glu Pro Arg Ile Asn Ser Tyr Arg Glu Phe Ala Asp Asp Val
        195                 200                 205
Leu Pro Arg Ile Arg Glu Asn Asn Tyr Asn Thr Val Gln Leu Met Ala
    210                 215                 220
Val Met Glu His Ser Tyr Tyr Ala Ser Phe Trp Tyr His Val Thr Lys
225                 230                 235                 240
Pro Phe Phe Ala Val Ser Ser Arg Ser Gly Ser Pro Glu Asp Leu Lys
                245                 250                 255
Tyr Leu Ile Asp Lys Ala His Ser Leu Gly Leu Asn Val Leu Met Asp
            260                 265                 270
Val Ile His Ser His Ala Ser Asn Asn Val Thr Asp Gly Leu Asn Gly
        275                 280                 285
Phe Asp Val Gly Gln Ser Ser Gln Gln Ser Tyr Phe His Ala Gly Asp
    290                 295                 300
Arg Gly Tyr His Lys Leu Trp Asp Ser Arg Leu Phe Asn Tyr Ala Asn
305                 310                 315                 320
Trp Lys Ser Ser Phe Leu Leu Ser Asn Leu Arg Trp Trp Leu Glu Glu
                325                 330                 335
Tyr Lys Phe Asp Gly Phe Arg Phe Asp Gly Val Thr Ser Met Leu Tyr
            340                 345                 350
His His His Gly Ile Asn Met Ala Phe Thr Gly Asp Tyr Asn Glu Tyr
        355                 360                 365
Phe Ser Glu Glu Thr Asp Val Asp Ala Val Val Tyr Leu Met Leu Ala
    370                 375                 380
Asn Ser Leu Val His Asp Ile Leu Pro Asp Ala Thr Asp Ile Ala Glu
385                 390                 395                 400
Asp Val Ser Gly Met Pro Gly Leu Gly Arg Pro Val Ser Glu Val Gly
                405                 410                 415
Ile Gly Phe Asp Tyr Arg Leu Ala Met Ala Ile Pro Asp Lys Trp Ile
            420                 425                 430
Asp Tyr Leu Lys Asn Lys Lys Asp Ser Glu Trp Ser Met Lys Glu Ile
        435                 440                 445
Ser Leu Asn Leu Thr Asn Arg Arg Tyr Thr Glu Lys Cys Val Ser Tyr
    450                 455                 460
Ala Glu Ser His Asp Gln Ser Ile Val Gly Asp Lys Thr Ile Ala Phe
465                 470                 475                 480
Leu Leu Met Asp Glu Glu Met Tyr Ser Ser Met Ser Cys Leu Thr Met
                485                 490                 495
Leu Ser Pro Thr Ile Glu Arg Gly Ile Ser Leu His Lys Met Ile His
            500                 505                 510
Phe Ile Thr Leu Ala Leu Gly Gly Glu Gly Tyr Leu Asn Phe Met Gly
        515                 520                 525
Asn Glu Phe Gly His Pro Glu Trp Ile Asp Phe Pro Arg Glu Gly Asn
    530                 535                 540
Gly Trp Ser Tyr Glu Lys Cys Arg Leu Thr Gln Trp Asn Leu Val Asp
545                 550                 555                 560
Thr Asn His Leu Arg Tyr Lys Phe Met Asn Ala Phe Asp Arg Ala Met
                565                 570                 575
Asn Leu Leu Asp Asp Lys Phe Ser Ile Leu Ala Ser Thr Lys Gln Ile
            580                 585                 590
Val Ser Ser Thr Asn Asn Glu Asp Lys Val Ile Val Phe Glu Arg Gly
        595                 600                 605
Asp Leu Val Phe Val Phe Asn Phe His Pro Glu Asn Thr Tyr Glu Gly
    610                 615                 620
Tyr Lys Val Gly Cys Asp Leu Pro Gly Lys Tyr Arg Val Ala Leu Asp
625                 630                 635                 640
Ser Asp Ala Thr Glu Phe Gly Gly His Gly Arg Val Gly His Asp Ala
                645                 650                 655
Asp Gln Phe Thr Ser Pro Glu Gly Ile Pro Gly Ile Pro Glu Thr Asn
            660                 665                 670
Phe Asn Asn Arg Pro Asn Ser Phe Lys Val Leu Ser Pro Pro His Thr
        675                 680                 685
Cys Val Val Tyr Tyr Arg Val Asp Glu Arg Gln Glu Glu Ser Asn Asn
    690                 695                 700
Pro Asn Leu Gly Ser Val Glu Glu Thr Phe Ala Ala Ala Asp Thr Asp
705                 710                 715                 720
Val Ala Arg Ile Pro Asp Val Ser Met Glu Ser Glu Asp Ser Asn Leu
                725                 730                 735
Asp Arg Ile Glu Asp Asn Ser Glu Asp Ala Val Asp Ala Gly Ile Leu
            740                 745                 750
Lys Val Glu Arg Glu Val Val Gly Asp Asn
        755                 760
 
           
             26 
             984 
             DNA 
             Triticum aestivum 
           
            26
atatgtatga tttcatggct ctggatagac cttcaactcc tcgcattgat cgtggcatag     60
cattacataa aatgatcagg cttgtcacca tgggtttagg tggcgaaggc tatcttaact    120
tcatgggaaa tgagtttggg catcctgaat ggatagattt tccaagaggt ccgcaaactc    180
ttccaaccgg caaagttctc cctggaaata acaatagtta tgataaatgc cgccgtagat    240
ttgatcttgg agatgcagat tttcttagat atcgtggtat gcaagagtcc gaccaggcaa    300
tgcagcatct tgaggaaaaa tatgggttta tgacatctga gcaccagtat gtttcacgga    360
aacatgagga agataaggtg atcatcttcg aaagaggaga tttggtattc gttttcaact    420
tccaccggag caatagcttt tttgactacc gtgttgggtg ttccaggcct gggaagtaca    480
aggtggcctt agactccgac gatgcactct ttggtggatt cagcaggctt gatcatgatg    540
tcgactactt cacaaccgaa catccgcatg acaacaggcc gcgctctttc tcggtgtaca    600
ctccgagcag aactgcggtc gtgtatgccc ttacagagta agaaccagca gctgcttgtt    660
acaaggcaaa gagagaactc cagagagctc gtggatcgtg agcgaagcga cgggcaacgg    720
cgcgaggctg ctctaagcgc catgactggg aggggatcgt gcctcttccc cagatgccag    780
gaggagcaga tggataggta gcttgttggt gagcgctcga aagaaaatgg acgggcctgg    840
gtgtttgtcg tgctgcacta ccctcctcct atcttgcaca ttcccggttg tctttgtaca    900
tataactaat aattgcccgt gcgctcaacg tgaacatata aatattctaa taataggtta    960
tcccgtgaaa aaaaaaaaaa aaaa                                           984
 
           
             27 
             977 
             DNA 
             Triticum aestivum 
           
            27
atatgtatga tttcatggct ctggatagac cttcaactcc tcgcattgat cgtggcatag     60
cattacataa aatgatcagg cttgtcacca tgggtttagg tggcgaaggc tatcttaact    120
tcatgggaaa tgagtttggg catcctgaat ggatagattt tccaagaggt ccgcaaactc    180
ttccaaccgg caaagttctc cctggaaata acaatagtta tgataaatgc cgccgtagat    240
ttgatcttgg agatgcagat tttcttagat atcgtggtat gcaagagttc gaccaggcaa    300
tgcagcatct tgaggaaaaa tatgggttta tgacatctga gcaccagtat gtttcacgga    360
aacatgagga agataaggtg atcatcttcg aaagaggaga tttggtattt gttttcaact    420
tccactggag caatagcttt tttgactacc gtgttgggtg ttccaagcct gggaagtaca    480
aggtggcctt agactccgac gatgcactct ttggtggatt cagcaggctt gatcatgatg    540
tcgactactt cacaaccgaa catccgcatg acaataggcc gcgctctttc ttggtgtaca    600
ctcctagcag aactgcggtc gtgtatgccc ttacagagta agaaccagca gcggcttgtt    660
acaaggcaaa gagagaactc cagggagctc gtggattgtg agcgaagcga cgggcaactg    720
cgtgaggctg ctctaagcgc catgactggg aggggatcgt gcctcttccc ctgatgccag    780
gaggatcaga tggataggta gcttgttggt gagcgctcga aagaaaatgg acgggcctgg    840
gtgtttgtcg tgctgcactt aaccctcctc ctatgttgca cattcccggg tgtttttgta    900
catataacta ataattgccc gtgcgcttca acatgaacat ataaatattc tatataaaaa    960
aaaaaaaaaa aaaaaaa                                                   977
 
           
             28 
             212 
             PRT 
             Triticum aestivum 
           
            28
Met Tyr Asp Phe Met Ala Leu Asp Arg Pro Ser Thr Pro Arg Ile Asp
1               5                   10                  15
Arg Gly Ile Ala Leu His Lys Met Ile Arg Leu Val Thr Met Gly Leu
            20                  25                  30
Gly Gly Glu Gly Tyr Leu Asn Phe Met Gly Asn Glu Phe Gly His Pro
        35                  40                  45
Glu Trp Ile Asp Phe Pro Arg Gly Pro Gln Thr Leu Pro Thr Gly Lys
    50                  55                  60
Val Leu Pro Gly Asn Asn Asn Ser Tyr Asp Lys Cys Arg Arg Arg Phe
65                  70                  75                  80
Asp Leu Gly Asp Ala Asp Phe Leu Arg Tyr Arg Gly Met Gln Glu Phe
                85                  90                  95
Asp Gln Ala Met Gln His Leu Glu Glu Lys Tyr Gly Phe Met Thr Ser
            100                 105                 110
Glu His Gln Tyr Val Ser Arg Lys His Glu Glu Asp Lys Val Ile Ile
        115                 120                 125
Phe Glu Arg Gly Asp Leu Val Phe Val Phe Asn Phe His Trp Ser Asn
    130                 135                 140
Ser Phe Phe Asp Tyr Arg Val Gly Cys Ser Lys Pro Gly Lys Tyr Lys
145                 150                 155                 160
Val Ala Leu Asp Ser Asp Asp Ala Leu Phe Gly Gly Phe Ser Arg Leu
                165                 170                 175
Asp His Asp Val Asp Tyr Phe Thr Thr Glu His Pro His Asp Asn Arg
            180                 185                 190
Pro Arg Ser Phe Leu Val Tyr Thr Pro Ser Arg Thr Ala Val Val Tyr
        195                 200                 205
Ala Leu Thr Glu
    210
 
           
             29 
             212 
             PRT 
             Zea mays 
           
            29
Met Tyr Asp Phe Met Ala Leu Asp Arg Pro Ser Thr Pro Thr Ile Asp
1               5                   10                  15
Arg Gly Ile Ala Leu His Lys Met Ile Arg Leu Ile Thr Met Gly Leu
            20                  25                  30
Gly Gly Glu Gly Tyr Leu Asn Phe Met Gly Asn Glu Phe Gly His Pro
        35                  40                  45
Glu Trp Ile Asp Phe Pro Arg Gly Pro Gln Arg Leu Pro Ser Gly Lys
    50                  55                  60
Phe Ile Pro Gly Asn Asn Asn Ser Tyr Asp Lys Cys Arg Arg Arg Phe
65                  70                  75                  80
Asp Leu Gly Asp Ala Asp Tyr Leu Arg Tyr His Gly Met Gln Glu Phe
                85                  90                  95
Asp Gln Ala Met Gln His Leu Glu Gln Lys Tyr Glu Phe Met Thr Ser
            100                 105                 110
Asp His Gln Tyr Ile Ser Arg Lys His Glu Glu Asp Lys Val Ile Val
        115                 120                 125
Phe Glu Lys Gly Asp Leu Val Phe Val Phe Asn Phe His Cys Asn Asn
    130                 135                 140
Ser Tyr Phe Asp Tyr Arg Ile Gly Cys Arg Lys Pro Gly Val Tyr Lys
145                 150                 155                 160
Val Val Leu Asp Ser Asp Ala Gly Leu Phe Gly Gly Phe Ser Arg Ile
                165                 170                 175
His His Ala Ala Glu His Phe Thr Ala Asp Cys Ser His Asp Asn Arg
            180                 185                 190
Pro Tyr Ser Phe Ser Val Tyr Thr Pro Ser Arg Thr Cys Val Val Tyr
        195                 200                 205
Ala Pro Val Glu
    210
 
           
             30 
             216 
             PRT 
             Zea mays 
           
            30
Met Tyr Asp Phe Met Ala Leu Asp Arg Pro Ser Thr Pro Arg Ile Asp
1               5                   10                  15
Arg Gly Ile Ala Leu His Lys Met Ile Arg Leu Val Thr Met Gly Leu
            20                  25                  30
Gly Gly Glu Gly Tyr Leu Asn Phe Met Gly Asn Glu Phe Gly His Pro
        35                  40                  45
Glu Trp Ile Asp Phe Pro Arg Gly Pro Gln Ser Leu Pro Asn Gly Ser
    50                  55                  60
Val Ile Pro Gly Asn Asn Asn Ser Phe Asp Lys Cys Arg Arg Arg Phe
65                  70                  75                  80
Asp Leu Gly Asp Ala Asp Tyr Leu Arg Tyr Arg Gly Met Gln Glu Phe
                85                  90                  95
Asp Gln Ala Met Gln His Leu Glu Gly Lys Tyr Glu Phe Met Thr Ser
            100                 105                 110
Asp His Ser Tyr Phe Ser Arg Lys His Glu Glu Asp Lys Val Ile Ile
        115                 120                 125
Phe Glu Arg Gly Asp Leu Val Phe Val Phe Asn Phe His Trp Ser Asn
    130                 135                 140
Ser Tyr Phe Asp Tyr Arg Val Gly Cys Phe Lys Pro Gly Lys Tyr Lys
145                 150                 155                 160
Ile Val Leu Asp Ser Asp Asp Gly Leu Phe Gly Gly Phe Ser Arg Leu
                165                 170                 175
Asp His Asp Ala Glu Tyr Phe Thr Ala Asp Trp Pro His Asp Asn Arg
            180                 185                 190
Pro Cys Ser Phe Ser Val Tyr Ala Pro Ser Arg Thr Ala Val Val Tyr
        195                 200                 205
Ala Pro Ala Gly Ala Glu Asp Glu
    210                 215
 
           
             31 
             217 
             DNA 
             Zea mays 
           
            31
tagcggggta ctcgttgctg cgcggcatgt gtggggctgt cgatgtgagg aaaaaccttc     60
ttccaaaacc ggcagatgca tgcatgcatg ctacaataag gttctgatac tttaatcgat    120
gctggaaagc ccatgcatct cgctgcgttg tcctctctat atatataaga ccttcaaggt    180
gtcaattaaa catagagttt tcgtttttcg ctttcct                             217
 
           
             32 
             686 
             PRT 
             Triticum aestivum 
           
            32
Met Leu Cys Leu Ser Ser Ser Leu Leu Pro Arg Pro Ser Ala Ala Ala
1               5                   10                  15
Asp Arg Pro Leu Pro Gly Ile Ile Ala Gly Gly Gly Gly Gly Lys Arg
            20                  25                  30
Leu Ser Val Val Pro Ser Val Pro Phe Leu Leu Arg Trp Leu Trp Pro
        35                  40                  45
Arg Lys Ala Lys Ser Lys Ser Phe Val Ser Val Thr Ala Arg Gly Asn
    50                  55                  60
Lys Ile Ala Ala Thr Thr Gly Tyr Gly Ser Asp His Leu Pro Ile Tyr
65                  70                  75                  80
Asp Leu Asp Leu Lys Leu Ala Glu Phe Lys Asp His Phe Asp Tyr Thr
                85                  90                  95
Arg Asn Arg Tyr Ile Glu Gln Lys His Leu Ile Glu Lys His Glu Gly
            100                 105                 110
Ser Leu Glu Glu Phe Ser Lys Gly Tyr Leu Lys Phe Gly Ile Asn Thr
        115                 120                 125
Glu His Gly Ala Ser Val Tyr Arg Glu Trp Ala Pro Ala Ala Glu Glu
    130                 135                 140
Ala Gln Leu Val Gly Asp Phe Asn Asn Trp Asn Gly Ser Gly His Lys
145                 150                 155                 160
Met Ala Lys Asp Asn Phe Gly Val Trp Ser Ile Arg Ile Ser His Val
                165                 170                 175
Asn Gly Lys Pro Ala Ile Pro His Asn Ser Lys Val Lys Phe Arg Phe
            180                 185                 190
Arg His His Gly Val Trp Val Glu Gln Ile Pro Ala Trp Ile Arg Tyr
        195                 200                 205
Ala Thr Val Thr Ala Ser Glu Ser Gly Ala Pro Tyr Asp Gly Leu His
    210                 215                 220
Trp Asp Pro Pro Ser Ser Glu Arg Tyr Val Phe Asn His Pro Arg Pro
225                 230                 235                 240
Pro Lys Pro Asp Val Pro Arg Ile Tyr Glu Ala His Val Gly Val Ser
                245                 250                 255
Gly Gly Lys Leu Glu Ala Gly Thr Tyr Arg Glu Phe Pro Asp Asn Val
            260                 265                 270
Leu Pro Cys Leu Arg Ala Thr Asn Tyr Asn Thr Val Gln Leu Met Gly
        275                 280                 285
Ile Met Glu His Ser Asp Ser Ala Ser Phe Gly Tyr His Val Thr Asn
    290                 295                 300
Phe Phe Ala Val Ser Ser Arg Ser Gly Thr Pro Glu Asp Leu Lys Tyr
305                 310                 315                 320
Leu Ile Asp Lys Ala His Ser Leu Gly Leu Arg Val Leu Met Asp Val
                325                 330                 335
Val His Ser His Ala Ser Asn Asn Val Ile Asp Gly Leu Asn Gly Tyr
            340                 345                 350
Asp Val Gly Gln Ser Ala His Glu Ser Tyr Phe Tyr Thr Gly Asp Lys
        355                 360                 365
Gly Tyr Asn Lys Met Trp Asn Gly Arg Met Phe Asn Tyr Ala Asn Trp
    370                 375                 380
Glu Val Leu Arg Phe Leu Leu Ser Asn Leu Arg Tyr Trp Met Asp Glu
385                 390                 395                 400
Phe Met Phe Asp Gly Phe Arg Phe Val Gly Val Thr Ser Met Leu Tyr
                405                 410                 415
Asn His Asn Gly Ile Asn Met Ser Phe Asn Gly Asn Tyr Lys Asp Tyr
            420                 425                 430
Ile Gly Leu Asp Thr Asn Val Asp Ala Phe Val Tyr Met Met Leu Ala
        435                 440                 445
Asn His Leu Met His Lys Leu Phe Pro Glu Ala Ile Val Val Ala Val
    450                 455                 460
Asp Val Ser Gly Met Pro Val Leu Cys Trp Pro Val Asp Glu Gly Gly
465                 470                 475                 480
Leu Gly Phe Asp Tyr Arg Gln Ala Met Thr Ile Pro Asp Arg Trp Ile
                485                 490                 495
Asp Tyr Leu Glu Asn Lys Gly Asp Gln Gln Trp Ser Met Ser Ser Val
            500                 505                 510
Ile Ser Gln Thr Leu Thr Asn Arg Arg Tyr Pro Glu Lys Phe Ile Ala
        515                 520                 525
Tyr Ala Glu Arg Gln Asn His Ser Ile Ile Gly Ser Lys Thr Met Ala
    530                 535                 540
Phe Leu Leu Met Glu Trp Glu Thr Tyr Ser Gly Met Ser Ala Met Asp
545                 550                 555                 560
Pro Asp Ser Pro Thr Ile Asp Arg Ala Ile Ala Leu Gln Lys Met Ile
                565                 570                 575
His Phe Ile Thr Met Ala Phe Gly Gly Asp Ser Tyr Leu Lys Phe Met
            580                 585                 590
Gly Asn Glu Tyr Met Asn Ala Phe Val Gln Ala Val Asp Thr Pro Ser
        595                 600                 605
Asp Lys Cys Ser Phe Leu Ser Ser Ser Asn Gln Thr Ala Ser His Met
    610                 615                 620
Asn Glu Glu Glu Lys Gly Ser Ala Leu Thr Lys Gly Tyr Thr His Leu
625                 630                 635                 640
Arg Ser Gly Cys Phe Asp Pro Ser Leu Pro Ser Thr Ser Ser Cys Ala
                645                 650                 655
Phe Leu Gly Pro Ser Asn Gln Ser Pro Phe Ser Lys Pro Phe Ile Gly
            660                 665                 670
Phe Pro Gly Cys Ile Phe Cys Cys Gly Leu Phe Lys Gly Glu
        675                 680                 685
 
           
             33 
             830 
             PRT 
             Triticum aestivum 
           
            33
Met Leu Cys Leu Thr Ala Pro Ser Cys Ser Pro Ser Leu Pro Pro Arg
1               5                   10                  15
Pro Ser Arg Pro Ala Ala Asp Arg Pro Gly Pro Gly Ile Ser Gly Gly
            20                  25                  30
Gly Asn Val Arg Leu Ser Ala Val Pro Ala Pro Ser Ser Leu Arg Trp
        35                  40                  45
Ser Trp Pro Arg Lys Ala Lys Ser Lys Phe Ser Val Pro Val Ser Ala
    50                  55                  60
Pro Arg Asp Tyr Thr Met Ala Thr Ala Glu Asp Gly Val Gly Asp Leu
65                  70                  75                  80
Pro Ile Tyr Asp Leu Asp Pro Lys Phe Ala Gly Phe Lys Glu His Phe
                85                  90                  95
Ser Tyr Arg Met Lys Lys Tyr Leu Asp Gln Lys His Ser Ile Glu Lys
            100                 105                 110
His Glu Gly Gly Leu Glu Glu Phe Ser Lys Gly Tyr Leu Lys Phe Gly
        115                 120                 125
Ile Asn Thr Glu Asn Asp Ala Thr Val Tyr Arg Glu Trp Ala Pro Ala
    130                 135                 140
Ala Met Asp Ala Gln Leu Ile Gly Asp Phe Asn Asn Trp Asn Gly Ser
145                 150                 155                 160
Gly His Arg Met Thr Lys Asp Asn Tyr Gly Val Trp Ser Ile Arg Ile
                165                 170                 175
Ser His Val Asn Gly Lys Pro Ala Ile Pro His Asn Ser Lys Val Lys
            180                 185                 190
Phe Arg Phe His Arg Gly Asp Gly Leu Trp Val Asp Arg Val Pro Ala
        195                 200                 205
Trp Ile Arg Tyr Ala Thr Phe Asp Ala Ser Lys Phe Gly Ala Pro Tyr
    210                 215                 220
Asp Gly Val His Trp Asp Pro Pro Ser Gly Glu Arg Tyr Val Phe Lys
225                 230                 235                 240
His Pro Arg Pro Arg Lys Pro Asp Ala Pro Arg Ile Tyr Glu Ala His
                245                 250                 255
Val Gly Met Ser Gly Glu Lys Pro Glu Val Ser Thr Tyr Arg Glu Phe
            260                 265                 270
Ala Asp Asn Val Leu Pro Arg Ile Lys Ala Asn Asn Tyr Asn Thr Val
        275                 280                 285
Gln Leu Met Ala Ile Met Glu His Ser Tyr Tyr Ala Ser Phe Gly Tyr
    290                 295                 300
His Val Thr Asn Phe Phe Ala Val Ser Ser Arg Ser Gly Thr Pro Glu
305                 310                 315                 320
Asp Leu Lys Tyr Leu Val Asp Lys Ala His Ser Leu Gly Leu Arg Val
                325                 330                 335
Leu Met Asp Val Val His Ser His Ala Ser Ser Asn Lys Thr Asp Gly
            340                 345                 350
Leu Asn Gly Tyr Asp Val Gly Gln Asn Thr Gln Glu Ser Tyr Phe His
        355                 360                 365
Thr Gly Glu Arg Gly Tyr His Lys Leu Trp Asp Ser Arg Leu Phe Asn
    370                 375                 380
Tyr Ala Asn Trp Glu Val Leu Arg Phe Leu Leu Ser Asn Leu Arg Tyr
385                 390                 395                 400
Trp Met Asp Glu Phe Met Phe Asp Gly Phe Arg Phe Asp Gly Val Thr
                405                 410                 415
Ser Met Leu Tyr Asn His His Gly Ile Asn Met Ser Phe Ala Gly Ser
            420                 425                 430
Tyr Lys Glu Tyr Phe Gly Leu Asp Thr Asp Val Asp Ala Val Val Tyr
        435                 440                 445
Leu Met Leu Ala Asn His Leu Met His Lys Leu Leu Pro Glu Ala Thr
    450                 455                 460
Val Val Ala Glu Asp Val Ser Gly Met Pro Val Leu Cys Arg Ser Val
465                 470                 475                 480
Asp Glu Gly Gly Val Gly Phe Asp Tyr Arg Leu Ala Met Ala Ile Pro
                485                 490                 495
Asp Arg Trp Ile Asp Tyr Leu Lys Asn Lys Asp Asp Leu Glu Trp Ser
            500                 505                 510
Met Ser Gly Ile Ala His Thr Leu Thr Asn Arg Arg Tyr Thr Glu Lys
        515                 520                 525
Cys Ile Ala Tyr Ala Glu Ser His Asp Gln Ser Ile Val Gly Asp Lys
    530                 535                 540
Thr Met Ala Phe Leu Leu Met Asp Lys Glu Met Tyr Thr Gly Met Ser
545                 550                 555                 560
Asp Leu Gln Pro Ala Ser Pro Thr Ile Asp Arg Gly Ile Ala Leu Gln
                565                 570                 575
Lys Met Ile His Phe Ile Thr Met Ala Leu Gly Gly Asp Gly Tyr Leu
            580                 585                 590
Asn Phe Met Gly Asn Glu Phe Gly His Pro Glu Trp Ile Asp Phe Pro
        595                 600                 605
Arg Glu Gly Asn Asn Trp Ser Tyr Asp Lys Cys Arg Arg Gln Trp Ser
    610                 615                 620
Leu Ala Asp Ile Asp His Leu Arg Tyr Lys Tyr Met Asn Ala Phe Asp
625                 630                 635                 640
Gln Ala Met Asn Ala Leu Asp Asp Lys Phe Ser Phe Leu Ser Ser Ser
                645                 650                 655
Lys Gln Ile Val Ser Asp Met Asn Glu Glu Lys Lys Ile Ile Val Phe
            660                 665                 670
Glu Arg Gly Asp Leu Val Phe Val Phe Asn Phe His Pro Ser Lys Thr
        675                 680                 685
Tyr Asp Gly Tyr Lys Val Gly Cys Asp Leu Pro Gly Lys Tyr Lys Val
    690                 695                 700
Ala Leu Asp Ser Asp Ala Leu Met Phe Gly Gly His Gly Arg Val Ala
705                 710                 715                 720
His Asp Asn Asp His Phe Thr Ser Pro Glu Gly Val Pro Gly Val Pro
                725                 730                 735
Glu Thr Asn Phe Asn Asn Arg Pro Asn Ser Phe Lys Ile Leu Ser Pro
            740                 745                 750
Ser Arg Thr Cys Val Ala Tyr Tyr Arg Val Glu Glu Lys Ala Glu Lys
        755                 760                 765
Pro Lys Asp Glu Gly Ala Ala Ser Trp Gly Lys Thr Ala Leu Gly Tyr
    770                 775                 780
Ile Asp Val Glu Ala Thr Gly Val Lys Asp Ala Ala Asp Gly Glu Ala
785                 790                 795                 800
Thr Ser Gly Ser Glu Lys Ala Ser Thr Gly Gly Asp Ser Ser Lys Lys
                805                 810                 815
Gly Ile Asn Phe Val Phe Leu Ser Pro Asp Lys Asp Asn Lys
            820                 825                 830
 
           
             34 
             818 
             PRT 
             Triticum aestivum 
           
            34
Met Ala Thr Phe Ala Val Ser Gly Trp Thr Leu Gly Val Ala Arg Pro
1               5                   10                  15
Ala Gly Ala Gly Gly Gly Leu Leu Pro Arg Ser Gly Ser Glu Arg Arg
            20                  25                  30
Gly Gly Val Asp Leu Pro Ser Leu Leu Leu Arg Lys Lys Asp Ser Ser
        35                  40                  45
Arg Ala Ala Ser Pro Gly Lys Val Leu Val Pro Asp Gly Glu Ser Asp
    50                  55                  60
Asp Leu Ala Ser Pro Ala Gln Pro Glu Glu Leu Gln Ile Pro Glu Asp
65                  70                  75                  80
Ile Glu Glu Gln Thr Ala Glu Val Asn Met Thr Gly Gly Thr Ala Glu
                85                  90                  95
Lys Leu Glu Ser Ser Glu Pro Thr Gln Gly Ile Val Glu Thr Ile Thr
            100                 105                 110
Asp Gly Val Thr Lys Gly Val Lys Glu Leu Val Val Gly Glu Lys Pro
        115                 120                 125
Arg Val Val Pro Lys Pro Gly Asp Gly Gln Lys Ile Tyr Glu Ile Asp
    130                 135                 140
Pro Thr Leu Lys Asp Phe Arg Ser His Leu Asp Tyr Arg Tyr Ser Glu
145                 150                 155                 160
Tyr Arg Arg Ile Arg Ala Ala Ile Asp Gln His Glu Gly Gly Leu Glu
                165                 170                 175
Ala Phe Ser Arg Gly Tyr Glu Lys Leu Gly Phe Thr Arg Ser Ala Glu
            180                 185                 190
Gly Ile Thr Tyr Arg Glu Trp Ala Pro Gly Ala His Ser Ala Ala Leu
        195                 200                 205
Val Gly Asp Phe Asn Asn Trp Asn Pro Asn Ala Asp Thr Met Thr Arg
    210                 215                 220
Asp Asp Tyr Gly Val Trp Glu Ile Phe Leu Pro Asn Asn Ala Asp Gly
225                 230                 235                 240
Ser Pro Ala Ile Pro His Gly Ser Arg Val Lys Ile Arg Met Asp Thr
                245                 250                 255
Pro Ser Gly Val Lys Asp Ser Ile Ser Ala Trp Ile Lys Phe Ser Val
            260                 265                 270
Gln Ala Pro Gly Glu Ile Pro Phe Asn Gly Ile Tyr Tyr Asp Pro Pro
        275                 280                 285
Glu Glu Glu Lys Tyr Val Phe Gln His Pro Gln Pro Lys Arg Pro Glu
    290                 295                 300
Ser Leu Arg Ile Tyr Glu Ser His Ile Gly Met Ser Ser Pro Glu Pro
305                 310                 315                 320
Lys Ile Asn Ser Tyr Ala Asn Phe Arg Asp Glu Val Leu Pro Arg Ile
                325                 330                 335
Lys Arg Leu Gly Tyr Asn Ala Val Gln Ile Met Ala Ile Gln Glu His
            340                 345                 350
Ser Tyr Tyr Ala Ser Phe Gly Tyr His Val Thr Asn Phe Phe Ala Pro
        355                 360                 365
Ser Ser Arg Phe Gly Thr Pro Glu Asp Leu Lys Ser Leu Ile Asp Arg
    370                 375                 380
Ala His Glu Leu Gly Leu Ile Val Leu Met Asp Ile Val His Ser His
385                 390                 395                 400
Ser Ser Asn Asn Thr Leu Asp Gly Leu Asn Gly Phe Asp Gly Thr Asp
                405                 410                 415
Thr His Tyr Phe His Gly Gly Pro Arg Gly His His Trp Met Trp Asp
            420                 425                 430
Ser Arg Leu Phe Asn Tyr Gly Ser Trp Glu Val Leu Arg Phe Leu Leu
        435                 440                 445
Ser Asn Ala Arg Trp Trp Leu Glu Glu Tyr Lys Phe Asp Gly Phe Arg
    450                 455                 460
Phe Asp Gly Val Thr Ser Met Met Tyr Thr His His Gly Leu Gln Met
465                 470                 475                 480
Thr Phe Thr Gly Asn Tyr Gly Glu Tyr Phe Gly Phe Ala Thr Asp Val
                485                 490                 495
Asp Ala Val Val Tyr Leu Met Leu Val Asn Asp Leu Ile His Gly Leu
            500                 505                 510
His Pro Asp Ala Val Ser Ile Gly Glu Asp Val Ser Gly Met Pro Thr
        515                 520                 525
Phe Cys Ile Pro Val Pro Asp Gly Gly Val Gly Leu Asp Tyr Arg Leu
    530                 535                 540
His Met Ala Val Ala Asp Lys Trp Ile Glu Leu Leu Lys Gln Ser Asp
545                 550                 555                 560
Glu Ser Trp Lys Met Gly Asp Ile Val His Thr Leu Thr Asn Arg Arg
                565                 570                 575
Trp Leu Glu Lys Cys Val Thr Tyr Ala Glu Ser His Asp Gln Ala Leu
            580                 585                 590
Val Gly Asp Lys Thr Ile Ala Phe Trp Leu Met Asp Lys Asp Met Tyr
        595                 600                 605
Asp Phe Met Ala Leu Asp Arg Pro Ser Thr Pro Arg Ile Asp Arg Gly
    610                 615                 620
Ile Ala Leu His Lys Met Ile Arg Leu Val Thr Met Gly Leu Gly Gly
625                 630                 635                 640
Glu Gly Tyr Leu Asn Phe Met Gly Asn Glu Phe Gly His Pro Glu Trp
                645                 650                 655
Ile Asp Phe Pro Arg Gly Pro Gln Thr Leu Pro Thr Gly Lys Val Leu
            660                 665                 670
Pro Gly Asn Asn Asn Ser Tyr Asp Lys Cys Arg Arg Arg Phe Asp Leu
        675                 680                 685
Gly Asp Ala Asp Phe Leu Arg Tyr His Gly Met Gln Glu Phe Asp Gln
    690                 695                 700
Ala Met Gln His Leu Glu Glu Lys Tyr Gly Phe Met Thr Ser Glu His
705                 710                 715                 720
Gln Tyr Val Ser Arg Lys His Glu Glu Asp Lys Val Ile Ile Phe Glu
                725                 730                 735
Arg Gly Asp Leu Val Phe Val Phe Asn Phe His Trp Ser Asn Ser Phe
            740                 745                 750
Phe Asp Tyr Arg Val Gly Cys Ser Arg Pro Gly Lys Tyr Lys Val Ala
        755                 760                 765
Leu Asp Ser Asp Asp Ala Leu Phe Gly Gly Phe Ser Arg Leu Asp His
    770                 775                 780
Asp Val Asp Tyr Phe Thr Thr Glu His Pro His Asp Asn Arg Pro Arg
785                 790                 795                 800
Ser Phe Ser Val Tyr Thr Pro Ser Arg Thr Ala Val Val Tyr Ala Leu
                805                 810                 815
Thr Glu
 
           
             35 
             813 
             DNA 
             Escherichia coli 
           
            35
gagctccgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg     60
tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg    120
tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg    180
ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc    240
cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg    300
aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca    360
tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc    420
aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg    480
atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg    540
cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata    600
tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg    660
accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat    720
gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct    780
tctatcgcct tcttgacgag ttcttctgag ctc                                 813
 
           
             36 
             7 
             PRT 
             Triticum aestivum 
           
            36
Met Asp Lys Asp Met Tyr Asp
1               5
 
           
             37 
             40 
             DNA 
             Artificial Sequence 
             
               Synthetic Oligonucleotide 
             
           
            37
aaggatccgt cgacatcgat aatacgactc actataggga                           40
 
           
             38 
             17 
             DNA 
             Artificial Sequence 
             
               Synthetic Oligonucleotide 
             
           
            38
aaggatccgt cgacatc                                                    17
 
           
             39 
             20 
             DNA 
             Artificial Sequence 
             
               Synthetic Oligonucleotide 
             
           
            39
atggacaagg atatgtatga                                                 20
 
           
             40 
             21 
             DNA 
             Artificial Sequence 
             
               Synthetic Oligonucleotide 
             
           
            40
ttttcttcac aacgccctgg g                                               21
 
           
             41 
             21 
             DNA 
             Artificial Sequence 
             
               Synthetic Oligonucleotide 
             
           
            41
tgtttgggag atcttcctcc c                                               21
 
           
             42 
             8 
             PRT 
             Triticum aestivum 
           
            42
Gly Val Trp Glu Ile Phe Leu Pro
1               5
 
           
             43 
             10 
             DNA 
             Artificial Sequence 
             
               Synthetic Oligonucleotide 
             
           
            43
cgggatcccg                                                            10
 
           
             44 
             34 
             DNA 
             Artificial Sequence 
             
               Synthetic Oligonucleotide 
             
           
            44
gatgagctcc gtttcgcatg attgaacaag atgg                                 34
 
           
             45 
             30 
             DNA 
             Artificial Sequence 
             
               Synthetic Oligonucleotide 
             
           
            45
gtcgagctca gaagaactcg tcaagaaggc                                      30
 
           
             46 
             27 
             DNA 
             Artificial Sequence 
             
               Synthetic Oligonucleotide 
             
           
            46
cccgacggcg aggatctcgt gctgacc                                         27
 
           
             47 
             35 
             DNA 
             Artificial Sequence 
             
               Synthetic Oligonucleotide 
             
           
            47
catgggtcac gacgagatcc tcgccgtcgg gcatg                                35
 
           
             48 
             30 
             DNA 
             Artificial Sequence 
             
               Synthetic Oligonucleotide 
             
           
            48
attaggtacc ggacttgctc cgctgtcggc                                      30
 
           
             49 
             30 
             DNA 
             Artificial Sequence 
             
               Synthetic Oligonucleotide 
             
           
            49
tataggtacc gaggcagcga cagagatgcc                                      30
 
           
             50 
             21 
             DNA 
             Artificial Sequence 
             
               Synthetic Oligonucleotide 
             
           
            50
agctgaatcc ggcggcatgg c                                               21
 
           
             51 
             21 
             DNA 
             Artificial Sequence 
             
               Synthetic Oligonucleotide 
             
           
            51
tgatagtctt gccagtcagg g                                               21
 
           
             52 
             2037 
             DNA 
             Zea mays 
           
            52
ttagctgaat ccggcggcat ggcaaggtag actgcagtgc agcgtgaccc ggtcgtgccc     60
ctctctagag ataatgagca ttgcatgtct aagttataaa aaattaccac atattttttt    120
tgtcacactt gtttgaagtg cagtttatct atctttatac atatatttaa actttactct    180
acgaataata taatctatag tactacaata atatcagtgt tttagagaat catataaatg    240
aacagttaga catggtctaa aggacaattg gtattttgac aacaggactc tacagtttta    300
tctttttagt gtgcatgtgt tctccttttt ttttttgcaa atagcttcac ctatataata    360
cttcatccat tttattagta catccattta gggtttaggg ttaatggttt ttatagacta    420
atttttttag tacatctatt ttattctatt ttagcctcta aattaagaaa actaaaactc    480
tattttagtt tttttattta ataatttaga tataaaatag aataaaataa agtgactaaa    540
aattaaacaa atacccttta agaaattaaa aaaactaagg aaacattttt cttgtttcga    600
gtagataatg ccagcctgtt aaacgccgtc gacgcagtct aacggacacc aaccagcgaa    660
ccagcagcgt cgcgtcgggc caagcgaagc agacggcacg gcatctctgt cgctgcctcg    720
gtaccggact tcgtccgctg tcggcatcca gaaattgcgt ggcggagcgg cagacgtgag    780
ccggcacggc aggcggcctc ctcctcctct cacggcaccg gcagctacgg gggattcctt    840
tcccaccgct ccttcgcttt cccttcctcg cccgccgtaa taaatagaca ccccctccac    900
accctctttc cccaacctcg tgttgttcgg agcgcacaca cacacaacca gatctccccc    960
aaatccaccc gtcggcacct ccgcttcaag gtacgccgct cgtcctcccc ccccctctct   1020
accttctcta gatcggcgtt ccggtccatg gttagggccc ggtagttcta cttctgttca   1080
tgtttgtgtt agatccgtgt ttgtgttaga tccgtgctgc tagcgttcgt acacggatgc   1140
gacctgtacg tcagacacgt tctgattgct aacttgccag tgtttctctt tggggaatcc   1200
tgggatggct ctagccgttc cgcagacggg atcgatttca tgattttttt tgtttcgttg   1260
catagggttt ggtttgccct tttcctttat ttcaatatat gccgtgcact tgtttgtcgg   1320
gtcatctttt catgcttttt tttgtcttgg ttgtgatgat gtggtctggt tgggcggtcg   1380
ttctagatcg gagtagaatt ctgtttcaaa ctacctggtg gatttattaa ttttggatct   1440
gtatgtgtgt gccatacata ttcatagtta cgaattgaag atgatggatg gaaatatcga   1500
tctaggatag gtatacatgt tgatgcgggt tttactgatg catatacaga gatgcttttg   1560
ttcgcttggt tgtgatgatg tggtgtggtt gggcggtcgt tcattcgttc tagatcggag   1620
tagaatactg tttcaaacta cctggtgtat ttattaattt tggaactgta tgtgtgtgtc   1680
atacatcttc atagttacga gtttaagatg gatggaaata tcgatctagg ataggtatac   1740
atgttgatgt gggttttact gatgcatata catgatggca tatgcagcat ctattcatat   1800
gctctaacct tgagtaccta tctattataa taaacaagta tgttttataa ttattttgat   1860
cttgatatac ttggatgatg gcatatgcag cagctatatg tggatttttt tagccctgcc   1920
ttcatacgct atttatttgc ttggtactgt ttcttttgtc gatgctcacc ctgttgtttg   1980
gtgttacttc tgcagatgca gatctttgtg aaaaccctga ctggcaagac tatcacc      2037
 
           
             53 
             1085 
             DNA 
             Triticum aestivum 
           
            53
atatgtatga tttcatggct ctggatggac cttcgactcc tgctattgat cgtggcatag     60
cattgcataa aatgattagg cttgtcacca tgggtttagg tggagagggt tatcttaact    120
ttatgggaaa tgagtttggg catcctgaat ggatagattt tccaagaggc ccacaagttc    180
ttccaactgg taagtttctc cctggaaata acaatagtta tgataaatgc cgtcgtagat    240
ttgatcttgg tgatgcagat tttcttaggt atcgtggtat gcaggagttt gatcaggcaa    300
tgcagcatct tgaggaaaaa tatgggttta tgacatctga gcaccagtat gtttctcgga    360
aacatgagga agataaggtg atcgtgtttg aaagagggga tttggtattt gttttcaact    420
tccactggag taatagcttt tttgactacc gtgttgggtg tttcaagcct gggaagtaca    480
aggtggtctt agactccgac gctggactct ttggtggatt tggtaggctt gatcatgctg    540
tcgagtactt cacttctgac tgtccgcatg acaacaggcc gcattctttc tcggtgtaca    600
ctcctagcag aacttgtgtt gtgtatgctc ttatggagta agcagcaagt gcagcatacg    660
ctgccgctgt tgttgctagt agcaaggaga gatcgtaggt cactacacca ggtgcagggt    720
ttgatatgga tttttgcttg agcgagtcct ggatgggcaa gacagcgtga tgctgtgtgt    780
gctcccaaat cgccatggcg ttgggagggg atcgtgcttc tttgtgttat gctttgtgga    840
tcaggatgga actcccctag gtagccttgt tggtgagcgc tcgaaagaaa atggacgggc    900
ctgggtgttt gcttaaattt tgttgcccta aaccctcgct cctatcttgt acattgccgg    960
tttagatagg gtttgttttt gtacattttt ttgatagtta atagacttat tgctcgtgtg   1020
cttgacgttt tacatgaaca tataaatatt ctaaataggt taaaaaaaaa aaaaaaaaaa   1080
aaaaa                                                               1085
 
           
             54 
             888 
             PRT 
             Triticum aestivum 
             
               PEPTIDE 
               (1)..(888) 
               Xaa = any amino acid 
             
           
            54
Met Leu Cys Leu Ser Xaa Ser Leu Leu Pro Arg Pro Ser Arg Ala Ala
1               5                   10                  15
Ala Asp Arg Pro Xaa Leu Pro Gly Ile Xaa Gly Gly Gly Xaa Xaa Arg
            20                  25                  30
Leu Ser Ala Val Pro Ala Pro Xaa Xaa Leu Arg Trp Xaa Trp Pro Arg
        35                  40                  45
Lys Ala Lys Ser Lys Ser Ser Val Pro Val Xaa Ala Xaa Xaa Xaa Xaa
    50                  55                  60
Ile Xaa Ala Thr Xaa Xaa Xaa Gly Val Xaa Xaa Leu Pro Ile Tyr Asp
65                  70                  75                  80
Leu Asp Pro Lys Leu Ala Xaa Phe Lys Xaa His Phe Asp Tyr Arg Xaa
                85                  90                  95
Xaa Xaa Tyr Xaa Xaa Gln Lys His Xaa Ile Glu Lys His Glu Gly Gly
            100                 105                 110
Leu Glu Glu Phe Ser Lys Gly Tyr Leu Lys Phe Gly Ile Asn Thr Glu
        115                 120                 125
Xaa Xaa Ala Xaa Val Tyr Arg Glu Trp Ala Pro Ala Ala Xaa Xaa Ala
    130                 135                 140
Gln Leu Val Gly Asp Phe Asn Asn Trp Asn Gly Ser Gly His Xaa Met
145                 150                 155                 160
Thr Lys Asp Asn Phe Gly Val Trp Ser Ile Arg Leu Ser Asn Asn Ala
                165                 170                 175
Asp Gly Ser Pro Ala Ile Pro His Gly Ser Lys Val Lys Phe Arg Phe
            180                 185                 190
Asp Thr Pro Ser Gly Val Trp Val Asp Ser Ile Pro Ala Trp Ile Lys
        195                 200                 205
Tyr Ala Val Gln Thr Ala Gly Glu Ile Gly Ala Pro Tyr Asp Gly Ile
    210                 215                 220
His Tyr Asp Pro Pro Ser Glu Glu Lys Tyr Val Phe Lys His Pro Gln
225                 230                 235                 240
Pro Lys Lys Pro Asp Ser Leu Arg Ile Tyr Glu Ala His Val Gly Met
                245                 250                 255
Ser Gly Pro Glu Pro Glu Ile Asn Thr Tyr Ala Glu Phe Arg Asp Glu
            260                 265                 270
Val Leu Pro Arg Ile Lys Ala Leu Gly Tyr Asn Ala Val Gln Leu Met
        275                 280                 285
Ala Ile Gln Glu His Ser Tyr Tyr Ala Ser Phe Gly Tyr His Val Thr
    290                 295                 300
Asn Phe Phe Ala Val Ser Ser Arg Ser Gly Thr Pro Glu Asp Leu Lys
305                 310                 315                 320
Ser Leu Ile Asp Lys Ala His Ser Leu Gly Leu Arg Val Leu Met Asp
                325                 330                 335
Val Val His Ser His Ala Ser Asn Asn Thr Leu Asp Gly Leu Asn Gly
            340                 345                 350
Phe Asp Val Gly Gln Gly Thr Asp Thr Ser Tyr Phe His Gly Gly Xaa
        355                 360                 365
Arg Gly His His Lys Met Trp Asp Ser Arg Leu Phe Asn Tyr Gly Asn
    370                 375                 380
Trp Glu Val Leu Arg Phe Leu Leu Ser Asn Ala Arg Tyr Trp Leu Asp
385                 390                 395                 400
Glu Phe Lys Phe Asp Gly Phe Arg Phe Asp Gly Val Thr Ser Met Leu
                405                 410                 415
Tyr Thr His His Gly Leu Asn Met Ser Phe Thr Gly Ser Tyr Lys Glu
            420                 425                 430
Tyr Phe Gly Leu Ala Thr Asp Val Asp Ala Val Val Tyr Leu Met Leu
        435                 440                 445
Ala Asn Asp Leu Ile His Gly Leu Xaa Pro Glu Ala Val Val Val Gly
    450                 455                 460
Glu Asp Val Ser Gly Met Pro Val Leu Cys Xaa Pro Val Asp Glu Gly
465                 470                 475                 480
Gly Val Gly Phe Asp Tyr Arg Leu Ala Met Ala Val Ala Asp Lys Trp
                485                 490                 495
Ile Asp Leu Leu Lys Asn Lys Asp Asp Xaa Trp Ser Met Gly Xaa Ile
            500                 505                 510
Val His Thr Leu Thr Asn Arg Arg Tyr Pro Glu Lys Cys Val Ala Tyr
        515                 520                 525
Ala Glu Ser His Asp Gln Ala Leu Val Gly Asp Lys Thr Ile Ala Phe
    530                 535                 540
Leu Leu Met Asp Lys Asp Met Tyr Asp Gly Met Ala Leu Xaa Xaa Pro
545                 550                 555                 560
Ser Ser Pro Thr Ile Asp Arg Gly Ile Ala Leu Gln Lys Met Ile His
                565                 570                 575
Leu Ile Thr Met Gly Leu Gly Gly Asp Gly Tyr Leu Asn Phe Met Gly
            580                 585                 590
Asn Glu Phe Gly His Pro Glu Trp Ile Asp Phe Pro Arg Gly Pro Gln
        595                 600                 605
Leu Pro Thr Gly Lys Pro Gly Asn Asn Asn Ser Tyr Asp Lys Cys Arg
    610                 615                 620
Arg Arg Phe Asp Leu Gly Asp Ala Asp Phe Leu Arg Tyr His Gly Met
625                 630                 635                 640
Asn Ala Phe Asp Gln Ala Met Gln His Leu Glu Asp Lys Tyr Gly Phe
                645                 650                 655
Leu Ser Ser Ser His Gln Tyr Val Ser Arg Lys Asn Glu Glu Asp Lys
            660                 665                 670
Val Ile Val Phe Glu Lys Gly Asp Leu Val Phe Val Phe Asn Phe His
        675                 680                 685
Trp Ser Asn Ser Tyr Phe Asp Tyr Arg Val Gly Cys Xaa Xaa Pro Gly
    690                 695                 700
Lys Tyr Lys Val Ala Leu Asp Ser Asp Ala Xaa Leu Phe Gly Gly Phe
705                 710                 715                 720
Gly Arg Xaa Xaa His Asp Xaa Asp His Phe Thr Ser Glu Xaa Xaa His
                725                 730                 735
Asp Asn Arg Pro Xaa Ser Phe Ser Val Leu Thr Pro Ser Arg Thr Cys
            740                 745                 750
Val Val Tyr Ala Pro Xaa Glu Xaa Ala Ala Xaa Val Thr Lys Xaa Tyr
        755                 760                 765
Xaa Xaa Xaa Xaa Xaa Leu Xaa Arg Xaa Xaa Gly Xaa Xaa Xaa Xaa Xaa
    770                 775                 780
Xaa Phe Leu Xaa Pro Xaa Lys Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Leu
785                 790                 795                 800
Xaa Xaa Xaa Xaa Xaa Xaa Pro Xaa Xaa Xaa Pro Xaa Ile Xaa Phe Xaa
                805                 810                 815
Xaa Xaa Gly Gly Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Lys Xaa
            820                 825                 830
Xaa Xaa Xaa Ala Val Xaa Xaa Xaa Xaa Xaa Ser Xaa Xaa Xaa Xaa Xaa
        835                 840                 845
Xaa Ile Leu Xaa Leu Xaa Xaa Xaa Xaa Ile Ile Xaa Xaa Xaa Xaa Xaa
    850                 855                 860
Xaa Xaa Leu Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa
865                 870                 875                 880
Lys Lys Lys Lys Lys Lys Lys Lys
                885
 
           
             55 
             1488 
             DNA 
             Zea mays 
           
            55
aagcttgcat gcctgcaggt cgactctaga ccaaatttca tggtagttgg gagcctaccc     60
agatttcatg attaactgtg ctattgaatt gttgaaaatg gttgtgtctg tcgtatccga    120
cggataacgg aaacccgtcc gaaattcaat gggcatgggc atagatatag atttgtaccc    180
actactagta tggtcgcagg cggatattgg ttgcaaccgc agatatagtt tcggggaaaa    240
ggattaggct cagctccatc cctagacccc acttgtgtgt gtgggggggt ctacccttca    300
aaaggaaaaa aaactacaca cagtgcatat aagaagatga atattccaaa attcagcagt    360
caagaagccc tgataaactg tctggcatag ctagtacttt atacacttca agaccaaaag    420
aaatcactaa gtacagattt tagtgactcg taagtacaga tatcatctta caaggcccag    480
cccagcgacc tattacacag cccgctcggg cccgcgacgt cgggacacat cttcttcccc    540
cttttggtga agctctgctc gcagctgtcc ggctgcttgg acgttcgtgt ggcagattca    600
tctgtcgtct cgtctcctgt gcttcctggg tagcttgtgc agtggagctg acatggtctg    660
agcaggctta aaatttgctc gtagacgagg agtaccagca cagcacgttg cggatttctc    720
tgcctgtgaa gtgcaacgtc taggattgtc acacgccttg gtcgcgtcga tgcggtggtg    780
agcagagcag caacagctgg gcggcccaaa gttggcttcc gtgtcttcgt cgtacgtacg    840
cgcgcgccgg ggacacgcag agagcggaga gcgagccgtg cacgggggag gtggtgtgca    900
agtgcagccg cgcgcccgcg cccgcgcccg gtgggcaacc caaaaagtac ccacgacaag    960
cgaaggcgcc aaagcgatcc aagctccgga acgcatcagc cacaagcagc cgagaaccga   1020
accggtgggc gacgcgtcgt gggacggacg cgggcgacgc ttccaaacgg ggccacgtac   1080
gccggcgtgt gcgtgcgtgc gtgcagacga caagccaagg cgaggcagcc cccgatcggg   1140
aaagcgtttt gggcgcgagc gctggcgtgc gggtcagtcg ctggtgcgca gtgccggggg   1200
gaacgggtat cgtggggggc gcggcggaga agagcgtggc gaggccgaga gcagcgcgcg   1260
gccgggtcac gcaacgcgcc ccacgtactg ccctccccct ccgcgcgcgc tagaaatacc   1320
gaggcctgga ccgggggccc ccccgtcaca tccatccatc gaccgatcga tcgccacagc   1380
caacaccacc cgccgaggcg acgcgacagc cgccaggagg aaggaataaa ctcactgcca   1440
gccagtgaag ggggagaagt gtactgctcc gtcgactcta gaggatcc                1488