Abstract
By reverse transcription-polymerase chain reaction (RT-PCR), cDNA fragment of wheat yellow mosaic virus (WYMV) RNA2 encoding 72 ku protein has been synthesized and cloned into plasmid pET30a(+) for overexpression in prokaryotic cells. BL21(DE3) pLys S ofE.coli transformed with the recombinant plasmid pETP72 containing the fragment has been induced to express the 72 ku protein on high level. The produced protein has been purified from sodium dodecyl sulfate-polyacrylamide gel (SDS-PAGE) for its antiserum preparation. In Western-blotting analysis, the antibodies reacted with the 72 ku protein expressed inE.coli.
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Yu, J. L., Yan, L. Y., Su, N. et al., Analysis of nucleotide sequence of wheat yellow mosaic virus genomic RNAs, Science in China, Ser. C. 1999, 42(5): 554.
Namba, S., Kashiwazaki, S., Lu, X. et al., Complete nucleotide sequence of wheat yellow mosaic bymovirus genomic RNAs, Arch. Virol., 1998, 143: 631.
Xing, Y. M., Yu, J. L., Liu, Y. et al., Identification, purification and serology of a fungus-borne wheat virus, Journal of Agricultural Biotechnology (in Chinese), 1994, 2(1): 91.
Sambrook, J., Fritsh, E. F., Maniatis, T., Molecular Cloning: a Laboratory Manual, 2nd ed., New York: Cold Spring Harbor Laboratory Press, 1989.
Hager, D. A., Burgess, R. R., Elution of proteins from sodiumdodecyl sulfate polyacrylamide gel, removal of sodium dodecyl sulfate and renaturation of enzymatic activity, Analytical Biochemistry, 1980, 109: 76.
Hames, B. D., Rickwood, D., Gel electrophoresis of proteins: a practical approach, 2nd ed., London: Oxford University Press, 1990.
Towbin, H., Staehelin, T., Gordon, J., Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications, Proc. Natl. Acad. Sci. USA, 1979, 76: 4350.
Murphy, F. A., Rychlik, W., Bishop, D. H. L. et al., Virus taxonomy: Classification and Nomenclature of Viruses (Sixth report of the international committee on taxonomy of viruses), Wien, New York: Springer-Verlag, 1995.
Schenk, P., Sohn, A., Adams, M. J. et al., Movement of bymoviruses and functions of RNA2-encoded proteins of barley yellow mosaic virus, Agronomie, 1995, 15: 387.
Dessens, J. T., Nguyen, M., Meyer, M., Primary structure and sequence analysis of RNA2 of a mechanically transmitted barley mild mosaic virus isolate: an evolutionary relationship betweenbymovirus andfurovirus, Arch. Virol., 1995, 140: 325.
Dessens, J. T., Meyer, M., Characterization of fungally and mechanically transmitted isolates of barley mild mosaic virus: two strains in competition, Virology, 1995, 212: 383.
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Xing, Y., Su, N., Li, D. et al. Over-expression of 72 ku protein of wheat yellow mosaic virus inE. coli and preparation of its antiserum. Chin.Sci.Bull. 45, 525–528 (2000). https://doi.org/10.1007/BF02887098
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DOI: https://doi.org/10.1007/BF02887098