Skip to main content
Log in

Over-expression of 72 ku protein of wheat yellow mosaic virus inE. coli and preparation of its antiserum

  • Notes
  • Published:
Chinese Science Bulletin

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. 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.

    Article  CAS  Google Scholar 

  2. Namba, S., Kashiwazaki, S., Lu, X. et al., Complete nucleotide sequence of wheat yellow mosaic bymovirus genomic RNAs, Arch. Virol., 1998, 143: 631.

    Article  PubMed  CAS  Google Scholar 

  3. 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.

    Google Scholar 

  4. Sambrook, J., Fritsh, E. F., Maniatis, T., Molecular Cloning: a Laboratory Manual, 2nd ed., New York: Cold Spring Harbor Laboratory Press, 1989.

    Google Scholar 

  5. 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.

    Article  PubMed  CAS  Google Scholar 

  6. Hames, B. D., Rickwood, D., Gel electrophoresis of proteins: a practical approach, 2nd ed., London: Oxford University Press, 1990.

    Google Scholar 

  7. 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.

    Article  PubMed  CAS  Google Scholar 

  8. 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.

    Google Scholar 

  9. 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.

    Article  Google Scholar 

  10. 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.

    Article  PubMed  CAS  Google Scholar 

  11. 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.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

About this article

Cite this article

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

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02887098

Keywords

Navigation