Maize chlorotic dwarf virus and resistance thereto

TECHNICAL FIELD
This invention relates to providing plants with resistance to maize chlorotic dwarf virus (MCDV) and viruses to which MCDV infection or resistance provides cross-resistance, including maize dwarf mosaic virus strain A (MDMV-A).
BACKGROUND OF THE INVENTION
Virus-induced diseases in agronomically important crops have cost farmers a great loss of income due to reduced yields. Traditionally, virus diseases have been controlled by breeding for host plant resistance or by controlling insects that transmit diseases. Chemical means of protection are not generally possible for most viruses, and where possible are not generally practical. It has been known for many years that viral symptoms can be reduced in virus-infected plants by prior inoculation with a mild strain of the same virus, a phenomena known as cross-protection, as described by Sequeira, L., Trends in Biotechnology, 2, 25 (1984). Cross-protection is considered successful if the disease symptoms of the superinfecting (the more virulent) virus can be delayed or suppressed. There are several disadvantages to applying this type of cross-protection to the field situation:
1) application of the mild strain virus to entire fields is usually not practical,
2) the mild strain might undergo mutation to a more highly virulent strain,
3) the protecting strain might interact synergistically with a non-related virus causing a severe pathogenic infection,
4) a protecting virus in one crop may be a severe pathogen in another crop, and
5) a protective strain may cause a significant loss of yield in itself.
One proposed solution to these disadvantages has been to introduce a single viral gene into the host plant genome to cross-protect, rather than infect with an intact virus. This single gene cross-protection strategy has already been proven successful using the coat protein gene from tobacco mosaic virus (TMV-CP). As reported by Abel, P. P., et. al., Science, 232, 738 (1986), transgenic tobacco plants, expressing TMV mRNA and coat protein (CP), demonstrated delayed or suppressed symptom development upon infection with TMV. TMV-CP transgenic tomato plants have been described by Nelson, R. S., et. al., Bio/Technology, 6, 403 (1988), to show evidence of protection from TMV as well as three strains of tomato mosaic virus (ToMV). Other approaches using DNA clones of viruses to engineer resistance include positive interference, as described by Golemboski et al. Proc. Natl. Acad. Sci. USA, 87, 6311 (1990) and Carr and Zaitlin, Mol. Pl. Microbe Inter., 4, 579 (1991); and antisense RNA, as described by Powell et al., Proc. Natl. Acad. Sci. USA, 86, 6949 (1989).
Numerous viruses exist for which resistance is desired. Maize chlorotic dwarf virus causes a somewhat variable mosaic or yellow streaking and occasional stunting in maize. Early infections can result in severe symptoms including premature death. The virus is spread by the blackfaced leafhopper (Graminella nigrifons). MCDV can overwinter in Johnsongrass (Sorghum halepense) and as a result has become a recurrent problem in areas where Johnsongrass is a common weed. Combined infections with maize dwarf mosaic virus can cause more severe symptoms although the syndrome is less well characterized than Corn Lethal Necrosis. Only limited success has been obtained to date in developing MCDV-resistant maize lines, due to the difficulties of selecting effidently for resistance to an obligately insect transmitted virus, as well as a lack of usable sources of resistance in agronomically useful maize lines. Thus, there is a continuing need for genes, plant transformation vectors, and transformed plant materials providing resistances to pathogenic viruses such as MCDV.
Unfortunately, while certain plant viruses, such as tobacco mosaic virus, have coat protein genes that are found on subgenomic RNA and are therefore relatively easy to identify and clone for use in engineered cross-protection, maize chlorotic dwarf virus belongs to a completely separate group, the only other (tentatively assigned) member of which is the spherical virus of the rice tungro disease (RTSV). In addition, MCDV has a number of unusual biological properties which make identification of an appropriate gene difficult. For example, all attempts to mechanically transmit MCDV have been unsuccessful. As another example, MCDV appears to be a phloem-restricted virus. MCDV also has three coat proteins, and it was not known whether expression of one protein would be sufficient to confer immunity or whether all three would need to be expressed. Nor was it known which protein would be the appropriate one to express if only one could be expressed. Further, the genome of MCDV has an unusual genome organization to provide for the expression of multiple coat proteins.

BRIEF DESCRIPTION OF THE DRAWING FIGURES
FIG. 1 is a schematic illustration of the manner in which the nucleic acid sequence of MCDV-type strain was obtained by sequencing overlapping cDNA clones.
FIG. 2 is an a schematic illustration of the unusual organization of the MCDV genome.

DISCLOSURE OF THE INVENTION
In the present invention, methods and materials are provided to isolate any or all of the three coat protein genes from maize chlorotic dwarf virus (MCDV). One or more of these genes (MCDV-CP.sub.x, where x is 1, 2, or 3) is then incorporated in an expression cassette designed for suitable expression in a plant cell system. The resulting transformation vector is then introduced into maize callus to provide cross-protection to MCDV-related viral infections. MCDV has a single, long RNA core having the sequence shown in SEQUENCE I.D. No. 4.
DESCRIPTION OF THE PREFERRED EMBODIMENTS
The present invention provides cDNA clones from the RNA genome of maize chlorotic dwarf virus which code substantially solely for the coat protein of the virus. These clones are incorporated into an expression cassette in which the cDNA clone is operably linked to plant or bacterial regulatory sequences which cause the expression of the cDNA clone in living plant or bacterial cells, respectively. It is important that the cloned gene have a start codon in the correct reading frame for the structural sequence. The resulting bacterial vectors can be readily inserted into bacteria for expression and characterization of the sequence. Accordingly, the present invention also provides bacterial cells containing as a foreign plasmid at least one copy of the foregoing bacterial expression cassette. In addition, the plant expression cassette preferably includes a strong constitutive promoter sequence at one end to cause the gene to be transcribed at a high level and a poly-A recognition sequence at the other end for proper processing and transport of the messenger RNA. An example of such a preferred (empty) expression cassette into which the cDNA of the present invention can be inserted is the pPHI414 plasmid developed by Beach et al. of Pioneer Hi-Bred International, Inc., Johnston, Iowa, as disclosed in U.S. patent application Ser. No. 07/785,648, filed Oct. 31, 1991. Highly preferred plant expression cassettes will be designed to include one or more selectable marker genes, such as kanamycin resistance or herbicide tolerance genes. The plant expression vectors of this invention can be inserted, using any convenient technique, including electroporation (in protoplasts), microprojectile bombardment, and microinjection, into cells from monocotyledonous or dicotyledonous plants, in cell or tissue culture, to provide transformed plant cells containing as foreign DNA at least one copy of the DNA sequence of the plant expression cassette. Preferably, the monocotyledonous species will be selected from maize, sorghum, wheat and rice, and the dicotyledonous species will be selected from soybean, alfalfa, tobacco and tomato. Using known techniques, protoplasts can be regenerated and cell or tissue culture can be regenerated to form whole fertile plants which carry and express the desired cDNA clone for MCDV coat protein. Accordingly, a highly preferred embodiment of the present invention is a transformed maize plant, the cells of which contain as foreign DNA at least one copy of the DNA sequence of an expression cassette of this invention.
Finally, this invention provides methods of imparting resistance to maize chlorotic dwarf virus to plants of a MCDV susceptible taxon, comprising the steps of:
a) culturing cells or tissues from at least one plant from the taxon,
b) introducing into the cells of the cell culture or tissue culture at least one copy of an expression cassette comprising a cDNA clone from the RNA genome of MCDV which codes substantially solely for the coat protein of the virus, operably linked to plant regulatory sequences which cause the expression of the cDNA clone in the cells, and
c) regenerating MCDV-resistant whole plants from the cell or tissue culture. Once whole plants have been obtained, they can be sexually or clonally reproduced in such manner that at least one copy of the sequence provided by the expression cassette is present in the cells of progeny of the reproduction.
Alternatively, once a single transformed plant has been obtained by the foregoing recombinant DNA method, conventional plant breeding methods can be used to transfer the coat protein gene and associated regulatory sequence via crossing and backcrossing. Such intermediate methods will comprise the further steps of
a) sexually crossing the MCDV resistant plant with a plant from the MCDV susceptible-taxon;
b) recovering reproductive material from the progeny of the cross; and
c) growing resistant plants from the reproductive material. Where desirable or necessary, the characteristics of the susceptible taxon can be substantially preserved by expanding this method to include the further steps of repetitively:
a) backcrossing the MCDV resistant progeny with MCDV susceptible plants from the susceptible taxon; and
b) selecting for expression of MCDV resistance among the progeny of the backcross,
until the desired percentage of the characteristics of the susceptible taxon are present in the progeny along with the gene imparting MCDV resistance.
By the term "taxon" herein is meant a unit of botanical classification of genus or lower. It thus includes genus, species, cultivars, varieties, variants, and other minor taxonomic groups which lack a consistent nomenclature.
It will also be appreciated by those of ordinary skill that the plant vectors provided herein can be incorporated into Agrobacterium tumefaciens or Agrobacterium rhizogenes, which can then be used to transfer the vector into susceptible plant cells, primarily from dicotyledonous species. Thus, this invention provides a method for imparting MCDV resistance in Agrobacterium-susceptible dicotyledonous plants in which the expression cassette is introduced into the cells by infecting the cells with Agrobacterium tumefaciens, a plasmid of which has been modified to include the plant expression cassette of this invention. The following description further exemplifies the compositions of this invention and the methods of making and using them. However, it will be understood that other methods, known by those of ordinary skill in the art to be equivalent, can also be employed.
1. Isolation and cloning of MCDV cDNA
The type strain of MCDV was maintained in the maize inbred Oh28 by transmission with the leafhopper G. nigrifrons and viral particles were isolated as previously described (Hunt et al., Phytopathology 78, 449 (1988)). MCDV particles were suspended in NETS (10 mM Tris, pH 7.5; 100 mM NaCl; 1 mM Na.sub.2 EDTA; 0.5% SDS) and extracted with 1:1 chloroform:phenol to isolate MCDV RNA.
First and second strand cDNA synthesis were by the method of Gubler and Hoffman, Gene 25, 263 (1983) utilizing cDNA synthesis kits (Amersham, Arlington Heights, Ill.). For the initial cDNA libraries, double-stranded cDNA was treated with EcoRI methylase, ligated to GGAATTCC EcoRI linkers, digested with EcoRI and separated from linkers by column fractionation. The cDNA was ligated to EcoRI-cleaved .sub.-- gt10 and EcORI-cleaved, phosphatased (CIP) .sub.-- gt11 phage arms. After packaging, the .sub.-- gt10 phage were plated on bacterial strain NM514 and screened for MCDV-specific inserts by filter plaque hybridization (Benton and Davis, Science 196, 180 (1977)), using .sup.32 P-labeled cDNA's random-primed from the MCDV genomic RNA. MCDV-positive phage were purified and the cDNA inserts subcloned into pUC119 (Vieira and Messing, Meth. Enzymol. 153, 3 (1987)) for further analysis. Hybridization positive clones from the initial gt10 library included: p3-13, p36-45, pH9, pK1, pG1, pC5 (FIG. 1). After packaging, the .sub.-- gt11 phage were plated on bacterial strain Y1090.sup.r- and screened with antisera to either intact MCDV virions or isolated, individual MCDV capsid proteins (Maroon, MS Thesis, Ohio State University (1989)) as described by Mierendorf, et al. Meth. Enzymol. 152,458 (1987). Positive phage clones were identified with antisera specific to either cp1 or cp2, and cDNA inserts from these phage were subcloned into pUC119. The anti-cp1-specific cDNA clone, p7C5, and the anti-cp2-specific cDNA clones, p7E6 and p7D7, (FIG. 1) were chosen for study. Analysis of initial cDNAs revealed that a number of clones terminated at identical EcoRI sites which were shown to be present in the viral sequence. This result indicated that the methylation of the initial cDNAs was incomplete. To obtain cDNAs to the rest of MCDV and to overlap the initial clones, two additional cDNA libraries were prepared, one primed with oligo-dT(12-18) and one random-primed. Double-stranded cDNA prepared as above was ligated to a 20/24 nt. blunt end/EcoRI adaptor (Amersham), and adaptor cDNAs were kinased and ligated to EcoRI-cleaved/phosphatased pUC119. Plasmid clone pdT2 (FIG. 1) was derived from the dT-primed library and plasmids pL142, pL221, and pL411 (FIG. 1) were derived from the random-primed library.
2. Sequencing of MCDV cDNA
Single-stranded DNA templates for sequencing were derived by superinfection with M13K07 of bacterial strain MV1190 containing the pUC119 based cDNAs (FIG. 1), cloned in both orientations, as described by McMullen et al., Nuc. Acids Res., 14, 4953 (1986) and Vieira and Messing, Meth. Enzymol. 153, 3 (1987). Ordered deletions from the full-length single-stranded templates were prepared by the method of Dale et al., Plasmid 13, 31 (1985). Dideoxynudeotide sequencing reactions with the Klenow fragment of Pol. I or Sequenase (U.S. Biochemicals, Cleveland, Ohio) were performed using .sup.35 S-dATP. Greater than 99% of the total sequence was obtained from both strands and the majority was read from three or more templates. The 5' sequence not contained on cDNA was obtained by direct RNA sequencing, using the sequencing primer 5'-GGTCTACTCACGGCACGCCA-3' (SEQUENCE I.D. NO. 3) with an RNA sequencing kit (Boehringer Mannheim, Indianapolis, Ind.) as recommended except that tailing of reaction products with dTTP by terminal deoxynucleotidyl transferase using the method of DeBorde et al., Anal. Biochem. 157,275 (1986) was added to improve resolution of final bases.
To obtain the amino-terminal protein sequence of MCDV capsid proteins, MCDV particles were disrupted in Laemmli loading buffer and the individual capsid protein separated on a 12.5%-4% Laemmli slab gel (Laemmli, Nature 227, 680 (1970)). The proteins were electrotransfered to Immobilon-P membrane (Millipore, Bedford, Mass.) using a 10 mM CAPS, pH 11.0; 10% MeOH transfer buffer, stained with Coomassie Blue R-250 for visualization and excised. Automated amino-terminal protein sequencing was performed by the Iowa State University Biochemistry Instrumentation Center (Ames, Iowa).
DNA and protein sequence analysis was performed using the IntelliGenetics (Mountain View, Calif.) molecular biology software on a Digital VAX 8250 located at the USDA-ARS-ASRR (Agricultural Systems Research Resource) Beltsville, Md.
The nucleic acid sequence of MCDV-type strain was obtained by sequencing overlapping cDNA clones (FIG. 1) that covered all but 13 nucleotides at the 5' terminus of MCDV. The 5' end sequence was obtained by direct RNA sequencing. Despite repeated attempts and the use of terminal transferase in the manner of DeBorde et al., Anal. Biochem., 157, 275 (1986) the first nucleotide could not be definitely determined. In part for this reason, the expressions "coding substantially for" and "coding substantially solely for" are used herein, and with regard to the use of the word "substantially" refer to sequences which code for no more than a few (five or less) amino acids greater or lesser on either end of the desired protein or proteins, or which have an equivalent number of nucleotide bases more or less than the native sequence.
The genomic RNA of MCDV-type (SEQUENCE I.D. NO. 4) was determined to be 11785 nucleotides long, exclusive of the poly-A tail at the 3' terminus. This sequence permits the construction of a DNA molecule which codes for the entire maize chlorotic dwarf virus, or any portion or functional unit thereof which is useful in conferring resistance to the virus when expressed in plant cells. Such resistance can readily be evaluated using routine testing methods such as those disclosed herein. Computer analysis of the sequence indicated a long open reading frame from nucleotide 456 to nucleotide 10826. The translation of this open reading frame would result in a protein of 3457 and no acids with a derived molecular weight of 388,890 daltons. The open reading frame begins with two AUG triplets, neither of which is in a particularly favorable context for initiation of translation when compared with the analyses of translation start sequences by Lutcke et al., EMBO J. 6, 43 (1987); and Kosak, J. Cell Biol. 108, 229 (1989) by the scanning model. In addition, there are 13 AUG triplets preceding the double AUG that starts the open reading frame. A long untranslated 3' leader containing multiple AUG triplets before the beginning of a very long reading frame is similar to the animal picornaviruses as described by Stanway, J. Gen. Virol. 71, 2483 (1990). Internal initiation at the AUG for the long open reading frame has been demonstrated to occur for a number of the animal picornaviruses as seen in Pelletier and Sonenberg, Nature, 334, 320 (1988) and Jang et al., J. Virol., 63, 1651 (1989). The mechanism for initiation of translation for MCDV has not been characterized.
The derived amino acid sequence of MCDV-type was compared to the Protein Identification Resource, Version 32 and the University of Geneva, Version 22, protein data banks for sequence similarity using the IFIND (IntelliGenetics) program based on the algorithm of Wilber and Lipman, Proc. Natl. Acad. Sci. USA, 80, 726 (1983). The highest similarity score was with the comovirus, cowpea mosaic virus (CPMV) as reported by Lomonossoff and Shanks, EMBO J., 2, 2253 (1983) and the second highest score was with the nepovirus, grapevine fanleaf virus (GFLV) as reported by Ritzenthaler et al., J. Gen. Virol., 72, 2357 (1991). For both viruses the region of similarity preceded and included the first conserved motif of RNA-dependent RNA polymerases as defined by Poch et al. EMBO J., 8, 3867 (1989). The IFIND program identified weaker similarity with additional nepoviruses and some of the animal picornaviruses. The conservation of protein sequence and gene order for the plant comoviruses, nepoviruses and potyviruses, and the animal picornaviruses is well documented by, inter alia, Agros et al., Nuc. Acids Res., 12, 7251 (1984); Goldbach, Ann. Rev. Phytopath., 24, 289 (1986); and Domier et al., Virology, 158, 20 (1987) and has led to the proposal of the picornavirus-like "supergroup". Two additional conserved protein regions involved in genome replication for picorna-like viruses are the NTP binding/helicase region, as described by Agros et al., above, and Gorbalenya et al., Nuc. Acids Res., 17, 4713 (1989) and the C-terminal region, cysteine active site of the 3C-like proteases, as also described by Agros et al., above, and by Grief et al., J. Gen. Virol., 69, 1517 (1988).
The electrophoresis of MCDV virions on denaturing protein gels reveals three structural proteins, designated cp1, cp2 and cp3 with molecular weights of 32.5 kd, 27 kd, and 24.5 kd; respectively. Antiserum specific to cp1 was used to screen a .sub.-- gt11 library to isolate the clone p7C5, and antiserum specific to cp2 was used to identify the cDNAs p7E6 and p7D7 (FIG. 1). This result indicated that an antigenic region of cp1 was located between 4063-4903 and an antigenic region of cp2 was located between 1815-2941. Automated amino-terminal sequencing was performed on each of the MCDV capsid proteins. The amino-terminus of cp2 was apparently blocked as no sequence was obtained. The 15 amino acids at the NH.sub.2 -terminus of cp3 were determined to be LQVASLTDIGELSSV, as shown in SEQUENCE I.D. NO. 2 and SEQUENCE I.D. NO. 6. This sequence is an exact match to the derived protein sequence encoded by nucleotides 3144-3188. Likewise, the 15 amino acids at the NH.sub.2 -terminus of cp1, VSLGRSFENGVLIGS, as shown in SEQUENCE I.D. NO. 5 and SEQUENCE I.D. NO. 7, are an exact match to the derived protein sequence encoded by nucleotides 3750-3794. Both proteins must be derived by proteolytic cleavage of the large polyprotein. The Gln/Leu cleavage at the NH.sub.2 -terminus of cp3 and Gln/Val cleavage at the NH.sub.2 -terminus of cp1 are dipeptide cleavage sites that may be used by animal picornavirus 3C proteases, according to Krausslich and Wimmer, Ann. Rev. Biochem., 57, 754 (1988), which could indicate that the 3C-similar region of the MCDV may function in capsid protein processing. Assuming that cp3 begins with the Leu at the Gln/Leu cleavage and ends with the GIn at the Gln/Val cleavage for cp1, cp3 would have a derived MW of 21,933, a little less than the 24.5 kd MW determined by SDS gel electrophoresis. Although protein sequence was not obtained for cp2, the position of clones p7E6 and p7D7, and the finding that protein fusions expressed from the pEX vector for the PstI fragments 2076-2619 and 2613-3149 reacted positively with cp2-specific antiserum (McMullen, unpublished), is consistent with cp2 preceding cp3 in the polyprotein similar to the order of vp2-vp3-vp1 for the animal picornaviruses. However, it is still not known if the coding region for cp2 immediately precedes cp3.
The overall genome structure of MCDV-type strain is shown in FIG. 2. MCDV genome organization resembled that of the animal picornaviruses, a single large polyprotein in which the capsid proteins are encoded 5' of the proteins presumed to be involved in genome replication. Depending on the exact location of cp2, the MCDV genome can encode up to 78 kd of protein 5' of the capsid proteins for which there are no corresponding animal picornavirus protein. This region may encode plant virus specific functions such as cell-to-cell movement or helper protein for insect transmission. Because MCDV is a phloem restricted virus, there is no evidence for a virus-encoded cell-to-cell movement protein. However, there is evidence for the presence of an insect transmission helper component in MCDV-infected plants according to Hunt et al., Phytopathology, 78, 449 (1988). The presence of plant-virus-specific proteins at the NH.sup.2 -terminus of the polyprotein would allow addition of these proteins without disruption of the cp proteins-replication functions genome structure typical of picornaviruses.
3. Design of the plasmid vector.
The gene MCDV coat protein 3 was placed under control of tandem cauliflower mosaic virus 35S promoters isolated from the 1841 strain of the virus, and a polyadenylation signal sequence obtained from the potato proteinase inhibitor II (Pin II) gene that exhibits enhancer-like activity. The chimeric gene also included a 79 bp sequence .OMEGA.' from the 5' leader region of tobacco mosaic virus (TMV) that functions as a translational enhancer; and a Zea mays alcohol dehydrogenase 1, intron 1 fragment (ADH) spanning nucleotides 119-672, trimmed to 557 bp with Bal 31 nuclease, which has been shown to function as an enhancer of gene expression in monocots. The plasmids were grown in E. coli and purified by the known polyethylene glycol precipitation method of Sambrook et al., Molecular Cloning, 1, 40 (1989). Purity was confirmed by electrophoretic analysis of the DNA fragments obtained after digestion with restriction endonucleases. The plasmid was designated pPHI1406 and the sequence is shown in SEQUENCE I.D. No. 1.
4. Preparation of the recipient organism.
Separately, an embryogenic cell suspension line 54-68-5 was established from immature embryos obtained from a cross between a line derived from the public inbred corn line B73 and a WX 1-9 translocation stock of public inbred corn line W23.
5. Transformation
Suspension cells from (4) were bombarded with 1 .mu.l aliquots of a 30 .mu.l mixture containing 10 .mu.g of purified plasmid DNA (5 .mu.g of the MCDV plasmid pPHI1406 (SEQUENCE I.D. No. 1), and 5 .mu.g of the same plasmid in which the BAR (Basta resistance) gene was substituted for the MCDV cp3 gene) precipitated onto 1 .mu.m tungsten particles as described by numerous articles including Klein, T. M., et al., 1988 (May) Bio/Technology 6:559-563; Klein, T. M., et al., 1988 (June) Proc. Natl. Acad. Sci. U.S.A. 85:4305-4309; T. M. Klein, et al., "Stable Genetic Transformation of Intact Applicant Nicotiana Cells by the Particle Bombardment Process", Proc. Natl. Acad. Sci. USA, Vol. 85, November 1988, pp. 8502-8505; D. T. Tomes, et al., "Transgenic Tobacco Plants and their Progeny Derived by Microprojectile Bombardment of Tobacco Leaves", Plant Molecular Biology, Vol. 14, No. 2, February, 1990, pp. 261-268, Kluwer Academic Publishers, BE; and M. C. Ross, et al., "Transient and Stable Transgenic Cells and Calli of Tobacco and Maize Following Microprojectile Bombardment", J. Cell. Biochem., Suppl. 13D, 27th Mar.-April 1989, P. 268, Abstract No. M. 149, Alan R. Liss, Inc. New York, U.S.; and plated onto selective medium containing 5 ppb phosphinothricin (Basta.TM.).
Following a prolonged period of selection and callus growth, regeneration was initiated by placing callus on a Murashige & Skoog medium modified by addition of 0.5 mg/12,4-D and 5 ppb Basta. Embryogenic callus was selected and transferred to medium lacking 2,4-D and kept in a lighted growth room. Germinated plantlets were placed in culture tubes and finally planted out into soil in pots in the greenhouse.
More than 150 R.sub.0 (recombinant) plants were obtained, representing twenty independent transformation events. Transformation was confirmed by PCR amplification of a DNA fragment spanning part of the MCDV coat protein gene and the CaMV promoter. Genomic DNA samples, in which a fragment of the expected size was successfully amplified were presumed to be transformed. These plants were pollinated with pollen from non-transgenic B73 plants and the resulting R.sub.1 seed was planted in a field trial under USDA supervision. The resulting plants exhibited a virus resistant phenotype, i.e., they survived and set seed under virus infection conditions in which non-transgenic plants died prematurely, as seen in the following table:
______________________________________Field Test Results Transgenic Control______________________________________Number of Plants 379 32Number of Harvestable Ears 52 0% Harvested vs. Total 13.7% 0%______________________________________
The screening was performed in a manner to insure maximum infection levels and severity. Thus, the level of resistance seen in this extreme test corresponds to effective, usable virus tolerance when the transformants of this invention are used trader normal farming conditions.
The MCDV resistance is a simply inherited, dominant trait and can, if desired, be introduced into other maize varieties by simple crossing or backcrossing. In addition to providing resistance to MCDV, this invention is also capable of conferring resistance to viruses to which plants obtain cross-resistance through infection by MCDV. In the field test described above, resistance to maize dwarf mosaic virus strain A (MDMV-A) was also observed. Accordingly, this invention provides resistance to that virus as well.
__________________________________________________________________________SEQUENCE LISTING(1) GENERAL INFORMATION:(iii) NUMBER OF SEQUENCES: 7(2) INFORMATION FOR SEQ ID NO: 1:(i) SEQUENCE CHARACTERISTICS:(A) LENGTH: 5033 base pairs(B) TYPE: nucleotide(C) STRANDEDNESS: double(D) TOPOLOGY: linear(ii) MOLECULE TYPE: synthetic DNA(A) DESCRIPTION: transformation plasmid pPHI1406(iii) HYPOTHETICAL: No(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 1:TCGCGCGTTTCGGTGATGACGGTGAAAACCTCTGACACATGCAGCTCCCG50GAGACGGTCACAGCTTGTCTGTAAGCGGATGCCGGGAGCAGACAAGCCCG100TCAGGGCGCGTCAGCGGGTGTTGGCGGGTGTCGGGGCTGGCTTAACTATG150CGGCATCAGAGCAGATTGTACTGAGAGTGCACCATATGCGGTGTGAAATA200CCGCACAGATGCGTAAGGAGAAAATACCGCATCAGGCGCCATTCGCCATT250CAGGCTGCGCAACTGTTGGGAAGGGCGATCGGTGCGGGCCTCTTCGCTAT300TACGCCAGCTGGCGAAAGGGGGATGTGCTGCAAGGCGATTAAGTTGGGTA350ACGCCAGGGTTTTCCCAGTCACGACGTTGTAAAACGACGGCCAGTGCCAA400GCTCAGATCTGAGCTTCTAGAAATCCGTCAACATGGTGGAGCACGACACT450CTCGTCTACTCCAAGAATATCAAAGATACAGTCTCAGAAGACCAAAGGGC500TATTGAGACTTTTCAACAAAGGGTAATATCGGGAAACCTCCTCGGATTCC550ATTGCCCAGCTATCTGTCACTTCATCAAAAGGACAGTAGAAAAGGAAGGT600GGCACCTACAAATGCCATCATTGCGATAAAGGAAAGGCTATCGTTCAAGA650TGCCTCTGCCGACAGTGGTCCCAAAGATGGACCCCCACCCACGAGGAGCA700TCGTGGAAAAAGAAGACGTTCCAACCACGTCTTCAAAGCAAGTGGATTGA750TGTGATGCTCTAGAAATCCGTCAACATGGTGGAGCACGACACTCTCGTCT800ACTCCAAGAATATCAAAGATACAGTCTCAGAAGACCAAAGGGCTATTGAG850ACTTTTCAACAAAGGGTAATATCGGGAAACCTCCTCGGATTCCATTGCCC900AGCTATCTGTCACTTCATCAAAAGGACAGTAGAAAAGGAAGGTGGCACCT950ACAAATGCCATCATTGCGATAAAGGAAAGGCTATCGTTCAAGATGCCTCT1000GCCGACAGTGGTCCCAAAGATGGACCCCCACCCACGAGGAGCATCGTGGA1050AAAAGAAGACGTTCCAACCACGTCTTCAAAGCAAGTGGATTGATGTGATA1100TCTCCACTGACGTAAGGGATGACGCACAATCCCACTATCCTTCGCAAGAC1150CCTTCCTCTATATAAGGAAGTTCATTTCATTTGGAGAGGACGAGCTGCAG1200CTTATTTTTACAACAATTACCAACAACAACAAACAACAAACAACATTACA1250ATTACTATTTACAATTACAGTCGACGGATCAAGTGCAAAGGTCCGCCTTG1300TTTCTCCTCTGTCTCTTGATCTGACTAATCTTGGTTTATGATTCGTTGAG1350TAATTTTGGGGAAAGCTTCGTCCACAGTTTTTTTTTCGATGAACAGTGCC1400GCAGTGGCGCTGATCTTGTATGCTATCCTGCAATCGTGGTGAACTTATGT1450CTTTTATATCCTTCACTACCATGAAAAGACTAGTAATCTTTCTCGATGTA1500ACATCGTCCAGCACTGCTATTACCGTGTGGTCCATCCGACAGTCTGGCTG1550AACACATCATACGATATTGAGCAAAGATCGATCTATCTTCCCTGTTCTTT1600AATGAAAGACGTCATTTTCATCAGTATGATCTAAGAATGTTGCAACTTGC1650AAGGAGGCGTTTCTTTCTTTGAATTTAACTAACTCGTTGAGTGGCCCTGT1700TTCTCGGACGTAAGGCCTTTGCTGCTCCACACATGTCCATTCGAATTTTA1750CCGTGTTTAGCAAGGGCGAAAAGTTTGCATCTTGATGATTTAGCTTGACT1800ATGCGATTGCTTTCCTGGACCCGTGCAGCTGCGGACGGATCCACCATGGC1850ACTGCAGGTGGCATCTCTTACAGACATAGGAGAATTGAGCAGTGTGGTTG1900CTACTGGTTCTTGGTCTACTACCTCGGCTACTAATTTGATGGAATTAAAC1950ATTCATCCCACCTCCTGTGCTATTCAGAACGGATTGATAACACAGACACC2000ATTGAGTGTTTTAGCTCATGCTTTTGCAAGGTGGAGAGGATCGTTGAAAA2050TTTCCATCATTTTCGGAGCGAGTTTGTTTACCCGAGGACGAATCTTAGCC2100GCTGCTGTGCCCGTTGCTAAGCGCAAAGGTACCATGAGCCTTGACGAGAT2150TAGTGGGTATCATAATGTTTGCTGCTTATTGAATGGTCAGCAAACTACAT2200TTGAATTGGAAATCCCATATTATTCTGTGGGCCAAGATTCTTTCGTGTAC2250CGTGATGCTCTTTTTGATATCTCTGCGCACGATGGGAATTTTATGATTAC2300TCGCTTGCATCTCGTGATACTGGATAAATTGGTAATGAGCGCTAATGCGA2350GCAACAGCATAAATTTTTCCGTGACTCTTGGACCAGGTTCTGATTTGGAA2400TTGAAATATCTTGCAGGAGTACATGGGCAGCGCATAGTCCGCGAGTTGAA2450GATGCAGTGATCAACCTAGACTTGTCCATCTTCTGGATTGGCCAACTTAA2500TTAATGTATGAAATAAAAGGATGCACACATAGTGACATGCTAATCACTAT2550AATGTGGGCATCAAAGTTGTGTGTTATGTGTAATTACTAGTTATCTGAAT2600AAAAGAGAAAGAGATCATCCATATTTCTTATCCTAAATGAATGTCACGTG2650TCTTTATAATTCTTTGATGAACCAGATGCATTTCATTAACCAAATCCATA2700TACATATAAATATTAATCATATATAATTAATATCAATTGGGTTAGCAAAA2750CAAATCTAGTCTAGGTGTGTTTTGCGAATTGCGGCCGCGATCTGGGGAAT2800TCGTAATCATGGTCATAGCTGTTTCCTGTGTGAAATTGTTATCCGCTCAC2850AATTCCACACAACATACGAGCCGGAAGCATAAAGTGTAAAGCCTGGGGTG2900CCTAATGAGTGAGCTAACTCACATTAATTGCGTTGCGCTCACTGCCCGCT2950TTCCAGTCGGGAAACCTGTCGTGCCAGCTGCATTAATGAATCGGCCAACG3000CGCGGGGAGAGGCGGTTTGCGTATTGGGCGCTCTTCCGCTTCCTCGCTCA3050CTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCAGCTCAC3100TCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCAGGAAA3150GAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCG3200CGTTGCTGGCGTTTTTCCATAGGCTCCGCCCCCCTGACGAGCATCACAAA3250AATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATA3300CCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTCCTGTTCCGACCC3350TGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCG3400CTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCG3450CTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCG3500CCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTA3550TCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGT3600AGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTA3650GAAGGACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGA3700AAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAGCGG3750TGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTC3800AAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAA3850AACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCAC3900CTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATAT3950ATGAGTAAACTTGGTCTGACAGTTACCAATGCTTAATCAGTGAGGCACCT4000ATCTCAGCGATCTGTCTATTTCGTTCATCCATAGTTGCCTGACTCCCCGT4050CGTGTAGATAACTACGATACGGGAGGGCTTACCATCTGGCCCCAGTGCTG4100CAATGATACCGCGAGACCCACGCTCACCGGCTCCAGATTTATCAGCAATA4150AACCAGCCAGCCGGAAGGGCCGAGCGCAGAAGTGGTCCTGCAACTTTATC4200CGCCTCCATCCAGTCTATTAATTGTTGCCGGGAAGCTAGAGTAAGTAGTT4250CGCCAGTTAATAGTTTGCGCAACGTTGTTGCCATTGCTACAGGCATCGTG4300GTGTCACGCTCGTCGTTTGGTATGGCTTCATTCAGCTCCGGTTCCCAACG4350ATCAAGGCGAGTTACATGATCCCCCATGTTGTGCAAAAAAGCGGTTAGCT4400CCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGCCGCAGTGTTATCA4450CTCATGGTTATGGCAGCACTGCATAATTCTCTTACTGTCATGCCATCCGT4500AAGATGCTTTTCTGTGACTGGTGAGTACTCAACCAAGTCATTCTGAGAAT4550AGTGTATGCGGCGACCGAGTTGCTCTTGCCCGGCGTCAATACGGGATAAT4600ACCGCGCCACATAGCAGAACTTTAAAAGTGCTCATCATTGGAAAACGTTC4650TTCGGGGCGAAAACTCTCAAGGATCTTACCGCTGTTGAGATCCAGTTCGA4700TGTAACCCACTCGTGCACCCAACTGATCTTCAGCATCTTTTACTTTCACC4750AGCGTTTCTGGGTGAGCAAAAACAGGAAGGCAAAATGCCGCAAAAAAGGG4800AATAAGGGCGACACGGAAATGTTGAATACTCATACTCTTCCTTTTTCAAT4850ATTATTGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATACATATTT4900GAATGTATTTAGAAAAATAAACAAATAGGGGTTCCGCGCACATTTCCCCG4950AAAAGTGCCACCTGACGTCTAAGAAACCATTATTATCATGACATTAACCT5000ATAAAAATAGGCGTATCACGAGGCCCTTTCGTC5033(2) INFORMATION FOR SEQ ID NO: 2:(i) SEQUENCE CHARACTERISTICS:(A) LENGTH: 45 bases(B) TYPE: nucleotide(C) STRANDEDNESS: single(D) TOPOLOGY: linear(ii) MOLECULE TYPE: viral RNA(A) DESCRIPTION: RNA codons for first 15 amino acids at 5'end of MCDV coat protein 3 (CP3)(iii) HYPOTHETICAL: No(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 2:CUGCAGGUGGCAUCUCUUACAGACAUAGGAGAAUUGAGCAGUGUG45LeuGlnValAlaSerLeuThrAspIleGlyAspLeuSerSerVal51015(2) INFORMATION FOR SEQ ID NO: 3:(i) SEQUENCE CHARACTERISTICS:(A) LENGTH: 20 bases(B) TYPE: nucleotide(C) STRANDEDNESS: single(D) TOPOLOGY: linear(ii) MOLECULE TYPE: synthetic DNA(A) DESCRIPTION: sequencing primer(iii) HYPOTHETICAL: No(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 3:GGTCTACTCACGGCACGCCA20(2) INFORMATION FOR SEQ ID NO: 4:(i) SEQUENCE CHARACTERISTICS:(A) LENGTH: 11785 bases(B) TYPE: nucleotide(C) STRANDEDNESS: single(D) TOPOLOGY: linear(ii) MOLECULE TYPE: viral RNA(iii) HYPOTHETICAL: No(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 4:NUGAAAAGGAGGGUAUAGAGAUACCCUUCAUAUAUUCUGCGGAUGGCGUG50CCGUGAGUAGACCUCGCGACGUUUCCCAGAGGAAAAUGGAAAUGGUCCAU100GUAACACCAGAUAUUUAUCUGGUUGAGGAACAUGGUUUAGUGGUAGAGAU150AAACUCAACUUUGUGUUGGACCCCGAUGCUGUGAAAAGUAAAUAAAGACA200AGGCCACUUAGCGAAGGAUAUUCGAAGUAGUGAUGAAAGGAAGUGCAAUA250AGUCAUGCCGUAAGUCGCAAUGCGCUAUAAGUCAUGCCGUAAGCCGCGUC300GCCUGGAUUUGCUAUUAGAAUGUCCCUAGCCGGUGAUAACCUUGAGUCCC350CGUCAUAGGACUACUUUUGUUUGCUUAGUAAUACAUUGGGACCACCCGCA400UGGAGCUCUGAGCCUACCAUACAUAGUACAUUUUCCGAGGGAUUGUCUUU450UGAUAAUGAUGCAGACAAACAACAACCAAAAUCCC485MetMetGlnThrAsnAsnAsnGlnAsnPro510ACUCAAGGAAGCAUUCCUGAGAACUCCUCACAAGAUCGCAACUUA530ThrGlnGlySerIleProGluAsnSerSerGlnAspArgAsnLeu152025GGAGUGCCCGCUGGAUAUUCUUUAAGCGUUGAGGACCCCUUCGGG575GlyValProAlaGlyTyrSerLeuSerValGluAspProPheGly303540AACCGGUCUGACUUUCAUAUCCCAGUGCACCAAAUCAUUCGGGAA620AsnArgSerAspPheHisIleProValHisGlnIleIleArgGlu455055GAGAUUGAUCGUCCAAAUUGGGUUCCUAUAUGUUCAAACGAUUUU665GluIleAspArgProAsnTrpValProIleCysSerAsnAspPhe606570CAUCUUAACAGUGAGGAUUAUUGUGAGGAGUGCGAAUCUGAACGG710HisLeuAsnSerGluAspTyrCysGluGluCysAspSerAspArg758085AUCAAAAAUUUCGAAAUAUUCAGAUCACAGAAUUUGAUUGACCAA755IleLysAsnPheAspIlePheArgSerGlnAsnLeuIleAspGln9095100CACCUAAAUCUCUGUACUGAUUCAAAGGAUUGUGAUCAUUUUUCU800HisLeuAsnLeuCysThrAspSerLysAspCysAspHisPheSer105110115UGUUUUUCCACGAGUACAAGUUGCAGAUUUUGCCCUUUUUGCUUA845CysPheSerThrSerThrSerCysArgPheCysProPheCysLeu120125130UUCAUUUUUAAUUUGGAUAAAUUUUACAAACAAAAUCUAUAUUUG890PheIlePheAsnLeuAspLysPheTyrLysGlnAsnLeuTyrLeu135140145AUUAGUCGUCAGGCUCUAGCUAGAUUGUUCCACGGAAGCGCCGAA935IleSerArgGlnAlaLeuAlaArgLeuPheHisGlySerAlaAsp150155160GAGUUACUCAGUAGAGCGAUUUUCUUUACGUAUAAUAUUUGUAUU980GluLeuLeuSerArgAlaIlePhePheThrTyrAsnIleCysIle165170175GAUGCAGAGGUGGUUGCUAAUAAUAGGAUUGGCUGUGAAUAUGUU1025AspAlaGluValValAlaAsnAsnArgIleGlyCysAspTyrVal180185190AAGUUGUUUCAUCCAGACCUUAGGCCUAGUAUUACGUCUCCCCCU1070LysLeuPheHisProAspLeuArgProSerIleThrSerProPro195200205UAUGCUAGUGAUUGGGUUAUGUGUGAUAAUGCUAAACAUCUUUUU1115TyrAlaSerAspTrpValMetCysAspAsnAlaLysHisLeuPhe210215220GAGUGUCUUGGCCUUGGUGACACGACCAGAGGACACCUAUAUGGA1160GluCysLeuGlyLeuGlyAspThrThrArgGlyHisLeuTyrGly225230235CUUAUUAGCGAGAAUGCAUAUUGGAACGCCACGUGCUCAAAAUGC1205LeuIleSerGluAsnAlaTyrTrpAsnAlaThrCysSerLysCys240245250GGAGCCUGUUGUCAGGGAGCAAAUGCCCGUACGGCGAUACCGAUA1250GlyAlaCysCysGlnGlyAlaAsnAlaArgThrAlaIleProIle255260265GUGAUGGCGUUGCAGUACUGCAGGGUGGAUGUGUAUUAUAGUGAG1295ValMetAlaLeuGlnTyrCysArgValAspValTyrTyrSerGlu270275280UACUAUUUAUACCACAUCUACGCUCCGGAAGAGAGAAUGAAGAUU1340TyrTyrLeuTyrHisIleTyrAlaProAspGluArgMetLysIle285290295GAUCAACAGACAGCACACUUGCUACACAGUAUAAUCCGAGGAGCA1385AspGlnGlnThrAlaHisLeuLeuHisSerIleIleArgGlyAla300305310CCAGCAGUGGAUUGCUCUGAGUUAUCUCAGGAGCCAAUUCACAGG1430ProAlaValAspCysSerGluLeuSerGlnGluProIleHisArg315320325AUGGUAAUGGAUAGCUCAAAGUUAGUGGCACUGGAUUCGACAAUC1475MetValMetAspSerSerLysLeuValAlaLeuAspSerThrIle330335340AGGCAUCCUAAGAGCCAAGGAAGUUUGCUCGAUUCAGAAUGCGAU1520ArgHisProLysSerGlnGlySerLeuLeuAspSerAspCysAsp345350355CAUGAGUUUAUUCUAAGAACGUCCCAUGGUAUCAAAAUACCGAUG1565HisGluPheIleLeuArgThrSerHisGlyIleLysIleProMet360365370AGUAAGUCUUUAUUUAUAUCAUUUCUUACCAUGGGAGCUUAUCAU1610SerLysSerLeuPheIleSerPheLeuThrMetGlyAlaTyrHis375400405GGGUAUGCUCAUGAUGAUCAGCAGGAGCAAAAUGCGAUAAUAUCU1655GlyTyrAlaHisAspAspGlnGlnGluGlnAsnAlaIleIleSer410415420UUUGGUGGGAUGCCCGGAGUCAAUUUGGCUUGUAACAAAAAUUUC1700PheGlyGlyMetProGlyValAsnLeuAlaCysAsnLysAsnPhe425430435CUGAGAAUGCAUAAGUUGUUUUAUUCUGGAAGUUUUAGGCGCAGA1745LeuArgMetHisLysLeuPheTyrSerGlySerPheArgArgArg440445450CCCCUGUUUAUGAGCCAAAUUCCCUCUACGAAUGCCACCGCUCAG1790ProLeuPheMetSerGlnIleProSerThrAsnAlaThrAlaGln455460465UCCGGUUUUAAUGAUGAAGAAUUCGAAAGAUUGAUGGCUGAAGAG1835SerGlyPheAsnAspAspAspPheAspArgLeuMetAlaAspGlu470475480GGUGUGCAUGUCAAAGUCGAGCGUCCAAUAGCAGAGAGGUUUGAU1880GlyValHisValLysValGluArgProIleAlaGluArgPheAsp485490500UAUGAGGACGUUAUUGAUAUUUACGAUGAGACCGACCACGACAGG1925TyrGluAspValIleAspIleTyrAspGluThrAspHisAspArg505510515ACACGAGCUCUAGGCCUUGGCCAAGUAUUCGGAGGUUUGCUCAAA1970ThrArgAlaLeuGlyLeuGlyGlnValPheGlyGlyLeuLeuLys520525530GGAAUUUCUCAUUGUGUAGAUAGCCUACAUAAGGUAUUUGAUUUC2015GlyIleSerHisCysValAspSerLeuHisLysValPheAspPhe535540545CCUCUGGACCUGGCCAUAGAAGCAGCUCAGAAAACUGGUGAUUGG2060ProLeuAspLeuAlaIleAspAlaAlaGlnLysThrGlyAspTrp550555560CUUGAAGGAAAUAAAGCUGCAGUAGAUGAAACUAAAAUUUGUGUG2105LeuAspGlyAsnLysAlaAlaValAspAspThrLysIleCysVal565670675GGCUGUCCCGAGAUUCAAAAAGAUAUGAUCAGUUUCCAGAAUGAA2150GlyCysProGluIleGlnLysAspMetIleSerPheGlnAsnAsp580585590ACAAAAGAAGCUUUUGAAUUAAUACGAUCAAGUAUAAAGAAGCUU2195ThrLysAspAlaPheAspLeuIleArgSerSerIleLysLysLeu595600605UCCGAGGGCAUUGACAAAAUCACGAAGAUGAAUGCUACGAACUUU2240SerGluGlyIleAspLysIleThrLysMetAsnAlaThrAsnPhe610615620GAACGAAUCCUAGACGGGAUUAAACCAAUCGAGAGCAGGUUGACA2285AspArgIleLeuAspGlyIleLysProIleGluSerArgLeuThr625630635GAACUUGAGAACAAGGCACCCGCUUCAGACAGCAAAGCCAUGGAA2330AspLeuGluAsnLysAlaProAlaSerAspSerLysAlaMetAsp640645650GCUCUGGUCCAGGCCGUGAAAGACUUGAAAAUCAUGAAAGAGGCG2375AlaLeuValGlnAlaValLysAspLeuLysIleMetLysGluAla655660665AUGCUCGAUCUAAAUCGAAGACUGAGCAAGCUGGAAGGAAAGAAA2420MetLeuAspLeuAsnArgArgLeuSerLysLeuAspGlyLysLys670675680AGUGAUGGCCAGACUACUGAAGGGACAGCGGGAGAGCAACAACCG2465SerAspGlyGlnThrThrAspGlyThrAlaGlyGluGlnGlnPro685690695AUCCCUAAGACUCCAACUCGAGUGAAGGCAAGACCAGUUGUGAAG2510IleProLysThrProThrArgValLysAlaArgProValValLys700705710CAAUCAGGAACGAUAAUGGUAAACGAAGAGAGCACAGAAACUUUC2555GlnSerGlyThrIleMetValAsnAspGluSerThrAspThrPhe715720725AGGGAUAAUGAGAGUCGAGUGACUGACCCUAACAGGAGCGAUAUG2600ArgAspAsnGluSerArgValThrAspProAsnArgSerAspMet730735740UUUGCUGCUGUUACUGCAGAAUACUUAGUUAAAUCGUUUACAUGG2645PheAlaAlaValThrAlaAspTyrLeuValLysSerPheThrTrp745750755AAAGUUUCUGAUGGACAAGAUAAAGUUUUGGCUGACCUUGAUUUA2690LysValSerAspGlyGlnAspLysValLeuAlaAspLeuAspLeu760765770CCUCAAGACUUAUGGAAAUCCAAUUCCCGAUUGAGUGAUAUCAUG2735ProGlnAspLeuTrpLysSerAsnSerArgLeuSerAspIleMet775780785GGGUAUUUCCAAUAUUAUGAUGCAACCGGAAUCACUUUUCGCAUA2780GlyTyrPheGlnTyrTyrAspAlaThrGlyIleThrPheArgIle790795800ACGACAACAUGUGUUCCUAUGCACGGUGGUACUUUAUGUGCUGCU2825ThrThrThrCysValProMetHisGlyGlyThrLeuCysAlaAla805810815UGGGAUGCUAAUGGUUGCGCUACACGACAAGGUAUAGCCACAACG2870TrpAspAlaAsnGlyCysAlaThrArgGlnGlyIleAlaThrThr820825830GUUCAGCUGACUGGUUUGCCCAAAACAUUUAUUGAAGCUCACAGC2915ValGlnLeuThrGlyLeuProLysThrPheIleAspAlaHisSer835840845UCAUCAGAAACGAUAAUCGUGGUAAAGAAUUCCAAUAUACAAUCC2960SerSerAspThrIleIleValValLysAsnSerAsnIleGlnSer850855860GCGAUUUGUCUAAGUGGAAGUGAGCACUCGUUUGGGAGAAUGGGA3005AlaIleCysLeuSerGlySerGluHisSerPheGlyArgMetGly865870875AUCCUGAAGAUCUGUUGCUUGAAUACGUUGAAUGCGCCAAAGGAA3050IleLeuLysIleCysCysLeuAsnThrLeuAsnAlaProLysAsp880885890GCUACACAGCAAGUGGCUGUGAACGUCUGGAUUAAGUUUGACGGA3095AlaThrGlnGlnValAlaValAsnValTrpIleLysPheAspGly895900905GUUAAAUUUCACGUUUAUUCUUUAAGGAAAAAUCCAGUCGUUUCG3140ValLysPheHisValTyrSerLeuArgLysAsnProValValSer910915920CAACUGCAGGUGGCAUCUCUUACAGACAUAGGAGAAUUGAGCAGU3185GlnLeuGlnValAlaSerLeuThrAspIleGlyAspLeuSerSer925930935GUGGUUGCUACUGGUUCUUGGUCUACUACCUCGGCUACUAAUUUG3230ValValAlaThrGlySerTrpSerThrThrSerAlaThrAsnLeu940945950AUGGAAUUAAACAUUCAUCCCACCUCCUGUGCUAUUCAGAACGGA3275MetAspLeuAsnIleHisProThrSerCysAlaIleGlnAsnGly955960965UUGAUAACACAGACACCAUUGAGUGUUUUAGCUCAUGCUUUUGCA3320LeuIleThrGlnThrProLeuSerValLeuAlaHisAlaPheAla970975980AGGUGGAGAGGAUCGUUGAAAAUUUCCAUCAUUUUCGGAGCGAGU3365ArgTrpArgGlySerLeuLysIleSerIleIlePheGlyAlaSer985990995UUGUUUACCCGAGGACGAAUCUUAGCCGCUGCUGUGCCCGUUGCU3410LeuPheThrArgGlyArgIleLeuAlaAlaAlaValProValAla100010051010AAGCGCAAAGGUACCAUGAGCCUUGACGAGAUUAGUGGGUAUCAU3455LysArgLysGlyThrMetSerLeuAspGluIleSerGlyTyrHis101510201025AAUGUUUGCUGCUUAUUGAAUGGUCAGCAAACUACAUUUGAAUUG3500AsnValCysCysLeuLeuAsnGlyGlnGlnThrThrPheAspLeu103010351040GAAAUCCCAUAUUAUUCUGUGGGCCAAGAUUCUUUCGUGUACCGU3545AspIleProTyrTyrSerValGlyGlnAspSerPheValTyrArg104510501055GAUGCUCUUUUUGAUAUCUCUGCGCACGAUGGGAAUUUUAUGAUU3590AspAlaLeuPheAspIleSerAlaHisAspGlyAsnPheMetIle106010651070ACUCGCUUGCAUCUCGUGAUACUGGAUAAAUUGGUAAUGAGCGCU3635ThrArgLeuHisLeuValIleLeuAspLysLeuValMetSerAla107510801085AAUGCGAGCAACAGCAUAAAUUUUUCCGUGACUCUUGGACCAGGU3680AsnAlaSerAsnSerIleAsnPheSerValThrLeuGlyProGly109010951100UCUGAUUUGGAAUUGAAAUAUCUUGCAGGAGUACAUGGGCAGCGC3725SerAspLeuAspLeuLysTyrLeuAlaGlyValHisGlyGlnArg110511101115AUAGUCCGCGAGUUGAAGAUGCAGGUUUCAUUGGGUCGGUCAUUU3770IleValArgGluLeuLysMetGlnValSerLeuGlyArgSerPhe112011251130GAGAAUGGAGUGCUUAUUGGUAGUGGCUUCGACGACUUGCUACAA3815GluAsnGlyValLeuIleGlySerGlyPheAspAspLeuLeuGln113511401145AGAUGGAGUCAUUUGGUGUCCAUGCCUUUUAAUGCAAAAGGAGAC3860ArgTrpSerHisLeuValSerMetProPheAsnAlaLysGlyAsp115011551160AGCGAUGAGAUCCAAGUCUUUGGCUAUAUCAUGACUGUUGCCCCG3905SerAspGluIleGlnValPheGlyTyrIleMetThrValAlaPro116511701175GCGUAUCGUUCCCUUCCAGUCCACUGCACGCUGCUAAGUUGGUUU3950AlaTyrArgSerLeuProValHisCysThrLeuLeuSerTrpPhe118011851190UCACAAUUAUUCGUGCAGUGGAAAGGUGGUAUAAAGUAUAGACUA3995SerGlnLeuPheValGlnTrpLysGlyGlyIleLysTyrArgLeu119512001205CACAUUGAUUCAGAAGAGCGCAGAUGGGGUGGAUUCAUCAAAGUU4040HisIleAspSerAspGluArgArgTrpGlyGlyPheIleLysVal121012151220UGGCAUGACCCAAAUGGCUCUUUGGAUGAAGGGAAAGAAUUUGCU4085TrpHisAspProAsnGlySerLeuAspAspGlyLysAspPheAla122512301235AAAGCGGAUAUUCUAUCGCCACCAGCCGGAGCUAUGGUUCGUUAU4130LysAlaAspIleLeuSerProProAlaGlyAlaMetValArgTyr124012451250UGGAACUAUUUAAAUGGAGACUUGGAGUUUACAGUACCAUUUUGU4175TrpAsnTyrLeuAsnGlyAspLeuGluPheThrValProPheCys125512601265GCUAGAACCAGUACGCUGUUCAUACCAAAAGCUAUGAUUGCCACC4220AlaArgThrSerThrLeuPheIleProLysAlaMetIleAlaThr127012751280GAUUCAAAGUCAUGGAUUCUGAACUACAACGGUACAUUGAAUUUC4265AspSerLysSerTrpIleLeuAsnTyrAsnGlyThrLeuAsnPhe128512901295GCGUACCAAGGAGUAGAUGACUUCACAAUUACAGUGGAAACAAGU4310AlaTyrGlnGlyValAspAspPheThrIleThrValAspThrSer130013051310GCAGCCGACGACUUUGAAUUUCACGUUCGAACAGUUGCACCCCGC4355AlaAlaAspAspPheAspPheHisValArgThrValAlaProArg131513201325GCUGGAAAGGUCAACGAAGCUUUUGCCAAAUUGGAGUACGCUUCU4400AlaGlyLysValAsnAspAlaPheAlaLysLeuGluTyrAlaSer133013351340GAUUUAAAGGAUAUCAAAGAAUCUCUGACAUCUUCCACUCGUUUG4445AspLeuLysAspIleLysAspSerLeuThrSerSerThrArgLeu134513501355AAAGGGCCUCAUUAUAAAACGAAAAUUACCUCAAUAGAGCCAAAU4490LysGlyProHisTyrLysThrLysIleThrSerIleGluProAsn136013651370AAAAUUGAUGAAAAUGAGUCCUCACGUGGUAAAGAUAACAAGUCA4535LysIleAspAspAsnGluSerSerArgGlyLysAspAsnLysSer137513801385AAUUCGAAAUUUGAGGACUUACUCAAUGCAACAGCUCAGAUGGAU4580AsnSerLysPheGluAspLeuLeuAsnAlaThrAlaGlnMetAsp139013951400UUUGAUCGAGCCACAGCGAACGUUGGGUGUGUGCCAUUCUCCAUU4625PheAspArgAlaThrAlaAsnValGlyCysValProPheSerIle140514101415GCAAAGACAGCAAAGGUGCUUUCGGAACGCGAGACGUGUAAGAAG4670AlaLysThrAlaLysValLeuSerAspArgGluThrCysLysLys142014251430AUGGCAGAUGUGUUAGAUUUCACACACUCAUGUUUGAACUUAGAC4715MetAlaAspValLeuAspPheThrHisSerCysLeuAsnLeuAsp143514401445AGUCAACCUGCGGCGGCAAGAUUAGCAGCGGCCAUUUCUCAAAUA4760SerGlnProAlaAlaAlaArgLeuAlaAlaAlaIleSerGlnIle150015051510GCACCUAUUAUGGAGAGCAUCGGUAGAACCACUCAAAGCGUAGAG4805AlaProIleMetGluSerIleGlyArgThrThrGlnSerValGlu151515201525GAAAAAUUGGCUUCUGUGGAUACAUUUAGGGACAAAAUCAUGGCU4850AspLysLeuAlaSerValAspThrPheArgAspLysIleMetAla153015351540CUAAUUUCAAACGUGCUUGGGGAUACUCUACCUGGACUGGCCAUU4895LeuIleSerAsnValLeuGlyAspThrLeuProGlyLeuAlaIle154515501555GCUGACUUCAAAAAAGGAAAAUAUGUGUGGGCCUCGUUCCUGACA4940AlaAspPheLysLysGlyLysTyrValTrpAlaSerPheLeuThr160016051610AUGAUAGCCGCUUGCGUAGUAGCUUGGGCUGCCACUAGCAAGAAA4985MetIleAlaAlaCysValValAlaTrpAlaAlaThrSerLysLys161516201625AGCUUCUUGAAAAGAUUUGCAGUGGUAGCUAUGAUAAUUUGGAGC5030SerPheLeuLysArgPheAlaValValAlaMetIleIleTrpSer163016351640CCAUUUCUCGCAAGUAAAAUAUGGGCGCUUGGUACAUGGAUUAGG5075ProPheLeuAlaSerLysIleTrpAlaLeuGlyThrTrpIleArg164516501655AAGAGCUGGAGUAAGCUUUGGCCUAAGUCAGACUCAUGCCGACAA5120LysSerTrpSerLysLeuTrpProLysSerAspSerCysArgGln166016651670CACUCUUUGGCAGGCCUGUGUGAAAGUGUGUUCACAUCAUUCAAG5165HisSerLeuAlaGlyLeuCysAspSerValPheThrSerPheLys167516801685GAUUUCCCUGACUGGUUUAAAUCAGGAGGAAUCACGAUUGUGACG5210AspPheProAspTrpPheLysSerGlyGlyIleThrIleValThr169016951700CAAGUUUGCACAGUAUUACUGACGAUAGUGAGUCUGAUUACACUU5255GlnValCysThrValLeuLeuThrIleValSerLeuIleThrLeu170517101715GGAACUAUACCAAGCACGAAACAAAAUGCUACGUUCGCAGACAAA5300GlyThrIleProSerThrLysGlnAsnAlaThrPheAlaAspLys172017251730UUUAAAGAAUUUGGUAACAUGAGCAGAGCUACAACGUCAAUAGCU5345PheLysAspPheGlyAsnMetSerArgAlaThrThrSerIleAla173517401745GCAGGUUACAAGACGAUAUCAGAGCUGUGUUCGAAAUUCACCAAU5390AlaGlyTyrLysThrIleSerGluLeuCysSerLysPheThrAsn175017551760UACUUGGCUGUAACCUUCUUUGGGGCGCAAGUUGAUGACGAUGCU5435TyrLeuAlaValThrPhePheGlyAlaGlnValAspAspAspAla176517701775UUCAAGGGUUUGGUAGCGUUCAACGUUAAGGAAUGGAUUCUUGAA5480PheLysGlyLeuValAlaPheAsnValLysAspTrpIleLeuAsp178017851790GUGAAAAACCUGUCUCUUGAGGAAAACAAAUUUAGUGGUUUUGGU5525ValLysAsnLeuSerLeuGluAspAsnLysPheSerGlyPheGly179518001805GGUGAUGAGCAUCUUGUCAAGGUUAGACAUUUAUAUGAUAAAUCU5570GlyAspGluHisLeuValLysValArgHisLeuTyrAspLysSer181018151820GUGGAAAUAACCUAUAAGUUGCUCCAGAAAAAUCGAGUUCCCAUU5615ValAspIleThrTyrLysLeuLeuGlnLysAsnArgValProIle182518301835GCUAUGCUUCCUAUCAUCCGAGACACGUGUAAGAAGUGCGAGGAU5660AlaMetLeuProIleIleArgAspThrCysLysLysCysGluAsp184018451850UUGCUAAACGAGAGUUAUACUUACAAAGGUAUGAAAACUCCGCGC5705LeuLeuAsnGluSerTyrThrTyrLysGlyMetLysThrProArg185518601865GUGGACCCAUUCUAUAUAUGCCUUUUUGGAGCACCUGGAGUUGGC5750ValAspProPheTyrIleCysLeuPheGlyAlaProGlyValGly187018751880AAGUCCACAGUGGCAUCGAUGAUUGUUGACGAUUUGUUGGAUGCU5795LysSerThrValAlaSerMetIleValAspAspLeuLeuAspAla188518901895AUGGGCGAACCUAAGGUUGAUAGGAUCUAUACGCGAUGCUGUUCU5840MetGlyAspProLysValAspArgIleTyrThrArgCysCysSer190019051910GAUCAAUAUUGGAGCAAUUAUCACCACGAGCCAGUUAUUUGUUAU5885AspGlnTyrTrpSerAsnTyrHisHisGluProValIleCysTyr191519201925GACGACUUGGGGGCAAUCAGCAGACCAGCGAGUUUAUCAGACUAU5930AspAspLeuGlyAlaIleSerArgProAlaSerLeuSerAspTyr193019351940GGGGAGAUAAUGGGAAUCAAAUCGAACAGACCAUACUCCCUACCU5975GlyGluIleMetGlyIleLysSerAsnArgProTyrSerLeuPro194519501955AUGGCUGCUGUUGAUGAGAAAGGAAGGCAUUGUUUAUCGCGAUAC6020MetAlaAlaValAspGluLysGlyArgHisCysLeuSerArgTyr196019651970CUCAUUGCUUGUACAAAUUUAACCCAUCUGGACGAUACGGGCGAU6065LeuIleAlaCysThrAsnLeuThrHisLeuAspAspThrGlyAsp197519801985GUGAAAACAAAGGAUGCCUACUAUCGCAGAAUCAAUGUCCCAGUG6110ValLysThrLysAspAlaTyrTyrArgArgIleAsnValProVal199019952000ACAGUGACGAGAGAAGUAACCGCCAUGAUGAACCCCGAGGACCCA6155ThrValThrArgAspValThrAlaMetMetAsnProGluAspPro200520102015ACUGAUGGACUACGUUUCACCGUGGAGCAAGUGCUUGAUGGAGGU6200ThrAspGlyLeuArgPheThrValGluGlnValLeuAspGlyGly202020252030AGAUGGAUUAAUGUUACUGAAAGCCGUCUCCUCAAUGGAAGGAUG6245ArgTrpIleAsnValThrAspSerArgLeuLeuAsnGlyArgMet203520402045CCAUUCAGGGCUGAAGAUCUCAUGAACAUGAACUACAGUUACUUU6290ProPheArgAlaAspAspLeuMetAsnMetAsnTyrSerTyrPhe205020552060AUGGAGUUUCUCAAGAUGUAUGCUGCUUUAUAUAUGGAAAAUCAA6335MetGluPheLeuLysMetTyrAlaAlaLeuTyrMetAspAsnGln206520702075AACAUGUUGGUGGCAAAAUUGAGAGGAACAGAGAUCCCAGAAUCA6380AsnMetLeuValAlaLysLeuArgGlyThrGluIleProAspSer208020852090CGUAGUUCAGAGAAUGAAGAACUUGAAUUCGAUUAUUUGGCUACA6425ArgSerSerGluAsnAspAspLeuAspPheAspTyrLeuAlaThr209521002105GCUCAGAUGGACCAUACAGUGACAUUUGGGGAACUAGUUACCAAA6470AlaGlnMetAspHisThrValThrPheGlyAspLeuValThrLys211021152120UUCAACUCGUAUAAGCUUACUGGGAAACAAUGGAACAAGAGGCUC6515PheAsnSerTyrLysLeuThrGlyLysGlnTrpAsnLysArgLeu212521302135UGGGAACUUGGAUGGACAUCUCUAGACGGAUGGAACACGAACAAG6560CysAspLeuGlyTrpThrSerLeuAspGlyTrpAsnThrAsnLys214021452150AUUAUGAGAUUCGACGAUCUAGUUGCCGGAUUCUGUGGUUGCUCA6605IleMetArgPheAspAspLeuValAlaGlyPheCysGlyCysSer215521602165AGGAAUGAGAAUUGCAAUUUUGACUUCUAUCAUCAGAGACUUCAA6650ArgAsnGluAsnCysAsnPheAspPheTyrHisGlnArgLeuGln217021752180GCAUGUUUGAACAAGAAAGGGUUUGCUCCCGCAUAUCAAUAUUUC6695AlaCysLeuAsnLysLysGlyPheAlaProAlaTyrGlnTyrPhe218521902195AACCUUCACAAGUUGAAUUCAGACACCCAGAAGACAGAGCUCAAG6740AsnLeuHisLysLeuAsnSerAspThrGlnLysThrGluLeuLys220022052210CUUAAAUGCGGGACAACUGCUGAAGAUUUAUUCAGACAAGCUGAC6785LeuLysCysGlyThrThrAlaAspAspLeuPheArgGlnAlaAsp221522202225UUGAUGGUCAUAUUCUCCUACCUCUUAUUUGUUGCGAGAAUUGGG6830LeuMetValIlePheSerTyrLeuLeuPheValAlaArgIleGly223022352240GUGAGUGGAUCUCAUGUGUGUCUGUCAUAUAACAUGUUGAACGUC6875ValSerGlySerHisValCysLeuSerTyrAsnMetLeuAsnVal224522502255AAGGAUGUCAAGGAUUUUGAGAUAUGCAGGGAGAACGUUCUUGAU6920LysAspValLysAspPheGluIleCysArgGluAsnValLeuAsp226022652270UUGUCCAGAAAAACUACAAUCGACGGUGAAGAAUGCUAUAUCUGG6965LeuSerArgLysThrThrIleAspGlyAspAspCysTyrIleTrp227522802285AAUUUUAUUUCUGAUAUCUUCCCACGCAUUGUGGCUAAGUACAAC7010AsnPheIleSerAspIlePheProArgIleValAlaLysTyrAsn229022952300UGUGUUGUGCUUAACGACGGAGAGAAGAGAUACAUCUUCGUGACU7055CysValValLeuAsnAspGlyGluLysArgTyrIlePheValThr230523102315GACAGCGCGCCCACUAGGAUCUUUCCCGAUUUGGCUUGGUCAGAU7100AspSerAlaProThrArgIlePheProAspLeuAlaTrpSerAsp232023252330CUUAUUUCCGGCAAGCAAGUUGUGAGUCCAAACAUUAUCAAAGUG7145LeuIleSerGlyLysGlnValValSerProAsnIleIleLysVal233523402345GCUGGAGAAACCAAGUCGAAAACCAUUGCCCCUCUGCUAGCAGAU7190AlaGlyAspThrLysSerLysThrIleAlaProLeuLeuAlaAsp235023552360UCCUACAAGGUUUUCAAGGAUCCGAAGGCAUGGCUUGAGAGGAAC7235SerTyrLysValPheLysAspProLysAlaTrpLeuGluArgAsn236523702375AAAGAAUUGAAAGCAGCUCUAGAAACAGAAGAAUAUAUCGCUCUC7280LysAspLeuLysAlaAlaLeuAspThrAspAspTyrIleAlaLeu238023852390CUCUUUGCUGUUGCAUGUGAAGCUGGUAGAUUCACUCAAAUUUUA7325LeuPheAlaValAlaCysAspAlaGlyArgPheThrGlnIleLeu239524002405GACAAACCUCCCAGUAGACGCAAGAUUUUAAAUAUGUCCGAAAGG7370AspLysProProSerArgArgLysIleLeuAsnMetSerAspArg241024152420UAUAAUGCAUAUAUUGAACAGGAAAAAGGGCUGAUUGGGAGACUU7415TyrAsnAlaTyrIleAspGlnAspLysGlyLeuIleGlyArgLeu242524302435UCUAAACCAGCAAAGAUAUGCUUAGCCAUAGGAACUGGAGUUGCG7460SerLysProAlaLysIleCysLeuAlaIleGlyThrGlyValAla244024452450AUCUUUGGGGCCCUAGCAGGCAUUGGAGUGGGUUUGUUUAAGCUG7505IlePheGlyAlaLeuAlaGlyIleGlyValGlyLeuPheLysLeu245524602465AUAGCUCACUUCAACAAAGAUGAAGAAGAGGUAGACGAAAUUGAA7550IleAlaHisPheAsnLysAspAspAspGluValAspAspIleAsp247024752480UUUGAUAUACUCUCCCCAGAGAUGAGCGGUUCGCACGAAUCCGGC7595PheAspIleLeuSerProGluMetSerGlySerHisAspSerGly248524902495CAACAUACCACGAGGUACGUCACGAAGGAGCGAGUUCCAUCCAAA7640GlnHisThrThrArgTyrValThrLysGluArgValProSerLys250025052510CCAGCAAGGAGGCAACAUGAAUUUGAUCUAAUGUUCGAUAAUCUA7685ProAlaArgArgGlnHisAspPheAspLeuMetPheAspAsnLeu251525202525CCCACUCCACAAGUUGAAGAGCUAAAGAGUGAGAUGACCUGCGCC7730ProThrProGlnValAspGluLeuLysSerGluMetThrCysAla253025352540AGUGCCAGUGAUGAGCAUAAGACUCAGUAUGUUAAAAGAAGAGUG7775SerAlaSerAspGluHisLysThrGlnTyrValLysArgArgVal254525502555GGACCUGUAAGCAAACGUAAGGAUGCUUCGGUAGCAGAAAUUAGU7820GlyProValSerLysArgLysAspAlaSerValAlaAspIleSer256025652570GGAGCUCAUGCGAGUGAUCAGCAUCAUACAGAAUACUUGAAAGCA7865GlyAlaHisAlaSerAspGlnHisHisThrAspTyrLeuLysAla257525802585CGCGUUCCACUCAUGAAAAGAAUAGCUACCAAAGAGAGCUAUGUU7910ArgValProLeuMetLysArgIleAlaThrLysGluSerTyrVal259025952600GUAACUUACGAUGACGAACCCAGCUCUCAUAUUUCCCUAGUUCGC7955ValThrTyrAspAspAspProSerSerHisIleSerLeuValArg260526102615AGGAUCCGACGUACACGACUGGCAAGAGCCAUCAAGCAAAUGGCA8000ArgIleArgArgThrArgLeuAlaArgAlaIleLysGlnMetAla262026252630GUCCUGGAGGACUUCCCAUCUACCUUGGAAGAGAUACGACUUUGG8045ValLeuGluAspPheProSerThrLeuAspGluIleArgLeuTrp263526402645AGACAAAACGCUGCAAAUAAAGGGGUUAUUGUUCCGAAGUACUCA8090ArgGlnAsnAlaAlaAsnLysGlyValIleValProLysTyrSer265026552660ACAAGUGGGAAAUUCUUCAGUGGCUUGUUGGAUGAUGAAGAAGAA8135ThrSerGlyLysPhePheSerGlyLeuLeuAspAspAspAspAsp266526702675GAACCUCAGAAUGUGAAUAUGUUGAACGAAGAGGACAUUGAGGUA8180AspProGlnAsnValAsnMetLeuAsnAspGluAspIleGluVal268026852690GAUAAGCGAAUGUUUGAGAAGAUUUCUGAGGUUAUAAGCGUGAUU8225AspLysArgMetPheGluLysIleSerGluValIleSerValIle269527002705CAACCCAGAAAGAAUGAGCUGGAAAGAAUGAUUGAGGAAGGCGUA8270GlnProArgLysAsnGluLeuAspArgMetIleGluAspGlyVal271027152720CACCACAAGGUCGUAAAGCAGGCAAGGGUUAACGACAAGGGCUUA8315HisHisLysValValLysGlnAlaArgValAsnAspLysGlyLeu272527302735GCCAAAGACCCCAACAUGGUGACUAUCUUGACGGACAAAUUAAUU8360AlaLysAspProAsnMetValThrIleLeuThrAspLysLeuIle274027452750AAUAUUAGUGCGGUGAUCGUCAAUUUAACGCCGACACGCCGGGCA8405AsnIleSerAlaValIleValAsnLeuThrProThrArgArgAla275527602765UACAUGAACGUGGUACGUCUUAUAGGCACUAUAGUUGUUUGCCCA8450TyrMetAsnValValArgLeuIleGlyThrIleValValCysPro277027752780GCCCACUACUUGGAAGCUUUAGAGGAAGGAGAUGAGCUGUAUUUC8495AlaHisTyrLeuAspAlaLeuGluAspGlyAspGluLeuTyrPhe278527902795AUUUGCUUCUCAUUGGUUAUCAAGCUCACUUUUGAUCCAAGUAGA8540IleCysPheSerLeuValIleLysLeuThrPheAspProSerArg280028052810GUGACUCUCGUGAAUAGCCAGCAGGAUUUGAUGGUUUGGGAUCUU8585ValThrLeuValAsnSerGlnGlnAspLeuMetValTrpAspLeu281528202825GGGAACAUGGUACCACCCUCAAUUGAUACUCUUAAAAUGAUACCU8630GlyAsnMetValProProSerIleAspThrLeuLysMetIlePro283028352840ACGCUUGAAGACUGGGAUCACUUUCAGGAUGGACCAGGAGCCUUU8675ThrLeuAspAspTrpAspHisPheGlnAspGlyProGlyAlaPhe284528502855GCUGUUACGAAAUAUAACUCGAAAUUCCCAACCAAUUAUAUCAAC8720AlaValThrLysTyrAsnSerLysPheProThrAsnTyrIleAsn286028652870ACACUGACUAUGAUUGAGAGGAUUAGGGCAAAUACUCAGAAUCCC8765ThrLeuThrMetIleGluArgIleArgAlaAsnThrGlnAsnPro287528802885ACGGGUUGUUAUUCCAUGAUGGGCUCCCAACAUACAAUCACCACA8810ThrGlyCysTyrSerMetMetGlySerGlnHisThrIleThrThr289028952900GGAUUGCGAUAUCAAAUGUUCUCUCUUGAUGGAUUCUGCGGUGGG8855GlyLeuArgTyrGlnMetPheSerLeuAspGlyPheCysGlyGly290529102915UUAAUCCUGAGAGCCAGCACAAACAUGGUGAGAAAGGUCGUCGGG8900LeuIleLeuArgAlaSerThrAsnMetValArgLysValValGly292029252930AUCCACGUUGCUGGAAGCCAGAAUCACGCUAUGGGAUAUGCAGAG8945IleHisValAlaGlySerGlnAsnHisAlaMetGlyTyrAlaGlu293529402945UGCCUUAUUGCAGAAGAUUUACGGGCUGCAGUGGCGAGAUUGGCG8990CysLeuIleAlaAspAspLeuArgAlaAlaValAlaArgLeuAla295029552960CUAGAUCCUAGAAGCACCAUCCAGGCAAGUCUGAAAGGUAGGAUU9035LeuAspProArgSerThrIleGlnAlaSerLeuLysGlyArgIle296529702975GAUGCUGUUUCUAAACAAUGUGGUUUAGACAGAGCUCUGGGUACG9080AspAlaValSerLysGlnCysGlyLeuAspArgAlaLeuGlyThr298029852990AUAGGAUGUCACGGGAAAGUUGCCUCUGAAGAUAUUACAAGUGCC9125IleGlyCysHisGlyLysValAlaSerAspAspIleThrSerAla299530003005GCCACGAAAACUUCCAUAAGAAAGUCAAGAAUACAUGGUCUAGUG9170AlaThrLysThrSerIleArgLysSerArgIleHisGlyLeuVal301030153020GGUGAGAUUAGAACUGAGCCUUCAAUUUUACACGCUCAUGAUCCC9215GlyGluIleArgThrGluProSerIleLeuHisAlaHisAspPro302530303035CGACUGCCUAAAGACAAGAUUGGGAAAUGGGACCCGGUUAUUGAG9260ArgLeuProLysAspLysIleGlyLysTrpAspProValIleGlu304030453050GCAUCAAUGAAGUAUGGUUCGAGAAUCACACCGUUCCCUGUAGAC9305AlaSerMetLysTyrGlySerArgIleThrProPheProValAsp305530603065CAAAUUCUGGAAGUGGAGGAUCAUCUUUCUAAAAUGUUGGCCAAU9350GlnIleLeuAspValGluAspHisLeuSerLysMetLeuAlaAsn307030753080UGUGAGAAUUCAAAAAACAAGCGGCAGGUUAAUAAUCUAGAAAUA9395CysGluAsnSerLysAsnLysArgGlnValAsnAsnLeuAspIle308530903095GGGAUUAAUGGAAUUGACCAGUCGGAUUAUUGGCAACAGAUAGAA9440GlyIleAsnGlyIleAspGlnSerAspTyrTrpGlnGlnIleAsp310031053110AUGGAUACUUCAAGUGGUUGGCCAUACGCUAAGCGUAAACCUGUU9485MetAspThrSerSerGlyTrpProTyrAlaLysArgLysProVal311531203125GGGGCAGCUGGAAAGAAAUGGCUAUUCGAGCAAGACGGCACAUAU9530GlyAlaAlaGlyLysLysTrpLeuPheGluGlnAspGlyThrTyr313031353140CCCUCCGGAAAACCUCGAUAUGUAUUUGGAGAUGCCGGGUUGAUU9575ProSerGlyLysProArgTyrValPheGlyAspAlaGlyLeuIle314531503155GAGAGCUAUAACUCGAUGCUUGGUGAGGCGAAGCAAGGCAUUAGU9620GluSerTyrAsnSerMetLeuGlyGluAlaLysGlnGlyIleSer316031653170CCCACUGUCGUCACAAUUGAGUGCGCAAAAGAUGAGAGGCGGAAG9665ProThrValValThrIleGluCysAlaLysAspGluArgArgLys317531803185CUUAAUAAGAUAUAUGAGAAACCCGCCACUCGGACGUUCACCAUA9710LeuAsnLysIleTyrGluLysProAlaThrArgThrPheThrIle319031953200CUGCCACCUGAGAUUAAUAUUUUAUUCAGGCAGUAUUUCGGAGAU9755LeuProProGluIleAsnIleLeuPheArgGlnTyrPheGlyAsp320532103215UUUGCAGCGAUGGUAAUGACAUGUAGAGCCAAGCUUUUCUGUCAA9800PheAlaAlaMetValMetThrCysArgAlaLysLeuPheCysGln322032253230GUUGGCAUCAACCCAGAGUCAAUGGAGUGGGGUGAUCUCAUGCUA9845ValGlyIleAsnProGluSerMetGluTrpGlyAspLeuMetLeu323532403245GGUCUAAAGGAGAAAUCAACUAAGGGAUUUGCAGGAGAUUAUUCG9890GlyLeuLysGluLysSerThrLysGlyPheAlaGlyAspTyrSer325032553260AAGUUCGAUGGAAUCGGAGACCCCCAGAUUUAUCAUUCAAUUACC9935LysPheAspGlyIleGlyAspProGlnIleTyrHisSerIleThr326532703275CAAGUAGUCAACAACUGGUAUAACGAUGGGGAAGAAAAUGCGACU9980GlnValValAsnAsnTrpTyrAsnAspGlyAspAspAsnAlaThr328032853290AUCAGGCAUGCUCUGAUAAGUAGCAUUAUACACAGGCGGGGCAUU10025IleArgHisAlaLeuIleSerSerIleIleHisArgArgGlyIle329533003305GUGAAAGAAUAUUUGUUCCAGUAUUGCCAGGGUAUGCCAUCAGGG10070ValLysAspTyrLeuPheGlnTyrCysGlnGlyMetProSerGly331033153320UUCGCCAUGACAGUGAUAUUCAAUUCGUUUAUGAACUAUUAUUAU10115PheAlaMetThrValIlePheAsnSerPheMetAsnTyrTyrTyr332533303335CUGUCUUUGGCCUGGAUGAAUCUGAUAAGUGCAUCCCCCCUUAGU10160LeuSerLeuAlaTrpMetAsnLeuIleSerAlaSerProLeuSer334033453350CCACAAGCUUCUUUGAGAUAUUUUGAUGAGUAUUGUAAGGUCAUU10205ProGlnAlaSerLeuArgTyrPheAspGluTyrCysLysValIle335533603365GUUUACGGUGAUGAUAAUAUUGUUGCCGUCAACGAAGAAUUCUUA10250ValTyrGlyAspAspAsnIleValAlaValAsnAspAspPheLeu337033753380GAGUACUAUAACUUGAGGCUUGUGGCAGGCUAUCUUAGUCAAUUU10295GluTyrTyrAsnLeuArgLeuValAlaGlyTyrLeuSerGlnPhe338533903395GGAGUAAGCUACACUGAUGACGCCAAGAACCCAAUAGAGAAGAGC10340GlyValSerTyrThrAspAspAlaLysAsnProIleGluLysSer340034053410GAACGAUAUGUGAAGAUAGAAGACGUUACGUUCUUAAAACGGCGA10385AspArgTyrValLysIleAspAspValThrPheLeuLysArgArg341534203425UGGGUGAGUCUUGGCGGUAGAGCUUCGAUGCUGUACAAAGCUCCG10430TrpValSerLeuGlyGlyArgAlaSerMetLeuTyrLysAlaPro343034353440CUUGACAAGGUUAGCAUUGAGGAAAGGCUUAACUGGAUCAGAGAG10475LeuAspLysValSerIleGluAspArgLeuAsnTrpIleArgGlu344534503455UGUGACGAUGGGGAACUAGCUCUGGUGCAGAACAUUGAAAGUGCU10520CysAspAspGlyAspLeuAlaLeuValGlnAsnIleAspSerAla346034653470CUGUACGAAGCUAGUAUUCAUGGCCACACAUAUUUUGGAGAGCUU10565LeuTyrAspAlaSerIleHisGlyHisThrTyrPheGlyGluLeu347534803485AAAGAUAAAAUUGCUAAAGCCUGUGAUGCAGUCAUGAUAACUAUG10610LysAspLysIleAlaLysAlaCysAspAlaValMetIleThrMet349034953500CCAAAUAUAAGAUAUAUUGACUGCCAGAGACGAUGGUGGACCUCC10655ProAsnIleArgTyrIleAspCysGlnArgArgTrpTrpThrSer350535103515AUGACUGGUGGGUAUCUUGAGCCGUCUGAUGUCACCAAACUUGUA10700MetThrGlyGlyTyrLeuGluProSerAspValThrLysLeuVal352035253530AGGCUUGUUGAGAAAGGACUACUAGACCCGAAAUCAGUAUGGAAA10745ArgLeuValGluLysGlyLeuLeuAspProLysSerValTrpLys353535403545GACCCAUUGUACAGAACCAACAAGUUGCUAUUCGACCUAUUGAGG10790AspProLeuTyrArgThrAsnLysLeuLeuPheAspLeuLeuArg355035553560GAGGUUAAGGCAGCACCCCUGGCCGCAUUUGUGGUCUAA10829GluValLysAlaAlaProLeuAlaAlaPheValVal35653570GUUACCCUUCUGACAAAAGGGCCUUGAACGGUUAUGGUUGAACAGAACUG10879UAAAAGGUGAGGACUAUAUAAGUUGUAGUACGGAUGAGAUUGAAAGAAAA10929UUGGGUCACUCCCAUUCCUUUAUUAGGAAGGAGUGAUACCUUUUGUGUAG10979AUCUCUACCCCGAAACUCUUGAACCCUCACACGUUUUGGAGUAACCAGUA11029CACCCUUUUAGGUGGACCCUCGACUAUAGAUCGAGACCAAGUAUUGACUU11079GGUGUUCACGUCUUGCCGGACGCAAAAUGGCACCCUUGUUUAGUGAUAUC11129AAGGUUACAAAUGUCACGCCCCACUAGUAAAAGUUUUGGUAUAUACGCAU11179UCGAACCGCCAAUGUAUACGUGUUUUCCCUUUUACUUUUUGUAUGUCGUC11229GUGGUGACGAGAUGCACGCCUGGUCAGCGGGGAAUAAGUUCACUAUAUGA11279ACAGACUCCGGCGAGCGAGACACGCUGUCGGCCUCGGGAGAGGGAACUAG11329CUCCAGGCACUUAAAUCCUGAAGUGUUAGAACUAAGCGUUUGAUCCUCCU11379CCGGGGGAAAGAGAACGCCAGUUCUUUAAGCCAUAACUCUAGUGAGUUGA11429AUCCUAUUCAUCCUUCUUAGGAUUAAGGAUUUCUGAAGUCUAUCAUGAAA11479AGUAGAUAGAAAGCAACACGUCAAUAACGUGGAACCUUUUCCGAGGAAGU11529AGGGUGCUUGUUCGAAAAUCAUGGUAGAUUCGGAAACAAUUUGCUUAGAG11579UGUGUCUUUUCGCGUUGGUAGUUCAACCGUUAGGGCUAGGCACACUUCUC11629CACGGGUUUGUGCUGCAGUAUUAAAUAUCAUUAAGGUACUGUGCUAUAGC11679GGAGAAAUUACAAAGCGUUGAACACAUUGACGAUGGGGCCCAAUGCGCAC11729CCGGAUGUGUUACGCACCGUUUUUCUCUGUGUCACUAUAGAUAAAAGUGG11779GGUAGC11785(2) INFORMATION FOR SEQ ID NO: 5:(i) SEQUENCE CHARACTERISTICS:(A) LENGTH: 45 bases(B) TYPE: nucleotide(C) STRANDEDNESS: single(D) TOPOLOGY: linear(ii) MOLECULE TYPE: viral RNA(A) DESCRIPTION: RNA codons for first 15 amino acids at5'end of MCDV coat protein 1 (CP1)(iii) HYPOTHETICAL: No(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 5:GUUUCAUUGGGUCGGUCAUUUGAGAAUGGAGUGCUUAUUGGUAGU45ValSerLeuGlyArgSerPheGluAsnGlyValLeuIleGlySer51015(2) INFORMATION FOR SEQ ID NO: 6:(i) SEQUENCE CHARACTERISTICS:(A) LENGTH: 15 amino acids(B) TYPE: amino acid(C) STRANDEDNESS: single(D) TOPOLOGY: linear(ii) MOLECULE TYPE: peptide(A) DESCRIPTION: first 15 amino acids of MCDV coat protein 3(iii) HYPOTHETICAL: No(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 6:LeuGlnValAlaSerLeuThrAspIleGlyAspLeuSerSerVal51015(2) INFORMATION FOR SEQ ID NO: 7:(i) SEQUENCE CHARACTERISTICS:(A) LENGTH: 15 amino acids(B) TYPE: amino acid(C) STRANDEDNESS: single(D) TOPOLOGY: linear(ii) MOLECULE TYPE: peptide(A) DESCRIPTION: first 15 amino acids of MCDV coat protein 1(iii) HYPOTHETICAL: No(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 7:ValSerLeuGlyArgSerPheGluAsnGlyValLeuIleGlySer51015__________________________________________________________________________

Maize chlorotic dwarf virus and resistance thereto

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Non-Patent Literature Citations (11)

Entry
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