Molecular Cloning of the Nucleoprotein Gene of Canine Distemper Virus Free

Abstract

SUMMARY

Messenger RNAs labelled in Vero cells infected with canine distemper virus were analysed by electrophoresis on 1.5% agarose gels containing 2-formaldehyde. Seven virus-specific RNA bands could be distinguished which were not sensitive to actinomycin D treatment and were confined to the polyadenylated RNA fraction. The most abundant virus-specific mRNA species had a molecular weight of 0.52 × 10 and its coding capacity was consistent with it being the mRNA for the most abundant virus-specific protein, the nucleoprotein. Polyadenylated RNA of this size class was purified by electrophoresis on a polyacrylamide gel and cloned into the I site of plasmid pBR322. A virus-specific clone obtained, clone 224, was then used to select messenger RNA from infected cells. The messenger RNA selected had a molecular weight of 0.52 × 10 and directed the synthesis of only the virus-specific nucleoprotein when used to stimulate a wheat germ cell-free system.

Keyword(s): CDV , cloning and NP gene
Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-65-3-549
1984-03-01
2024-03-28
Loading full text...

Full text loading...

/deliver/fulltext/jgv/65/3/JV0650030549.html?itemId=/content/journal/jgv/10.1099/0022-1317-65-3-549&mimeType=html&fmt=ahah

References

  1. Barrett T., Gorman N. T., Patterson R. C., Mahy B. W. J. 1983; Molecular studies on canine distemper virus replication. In Molecular Biology of Negative Strand Viruses Edited by Bishop D. H. L., Compans R. W. New York & London: Academic Press; (in press)
    [Google Scholar]
  2. Bellini W. J., Englund G., Richardson C. D., Hogan R. N., Rozenblatt S., Meyers C. A., Lazzarini R. A. 1983; Positive identification and molecular cloning of the phosphoprotein (P) of measles virus. In Molecular Biology of Negative Strand Viruses Edited by Bishop D. H. L., Compans R. W. New York & London: Academic Press; (in press)
    [Google Scholar]
  3. Birnboim H. C., Doly Y. 1979; A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Research 7:1513–1523
    [Google Scholar]
  4. Caton A. J. 1980 Sequence analysis of influenza virus RNA Ph.D. thesis University of Cambridge:
    [Google Scholar]
  5. Cohen S. N., Chang A. C. Y., Hsu L. 1972; Nonchromosome antibiotic resistance in bacteria: genetic transformation of E. coli by R-factor DNA. Proceedings of the National Academy of Sciences, U,. S,. A 69:2110–2114
    [Google Scholar]
  6. Floyd R. W., Stone M. P., Joklik W. K. 1974; Separation of single-stranded ribonucleic acids by acrylamide-agarose-urea-gel electrophoresis. Analytical Biochemistry 59:599–609
    [Google Scholar]
  7. Glass S. E., McGeoch D., Barry R. D. 1975; Characterisation of the mRNA of influenza virus. Journal of Virology 16:1435–1443
    [Google Scholar]
  8. Gorecki M., Rozenblatt S. 1980; Cloning of DNA complementary to the measles virus mRNA encoding nucleocapsid protein. Proceedings of the National Academy of Sciences, U,. S,. A 77:3686–3690
    [Google Scholar]
  9. Inglis M. M., Darby G. 1981; Adenovirus late sequences linked to herpes simplex virus thymidine kinase may be introduced into eukaryotic cells and transcribed. Nucleic Acids Research 9:5569–5585
    [Google Scholar]
  10. Katz L., Kingsbury D. T., Helinski D. R. 1973; Stimulation by cyclic adenosine monophosphate of plasmid deoxyribonucleic acid replication and catabolite repression of the plasmid deoxyribonucleic acid-protein relaxation complex. Journal of Bacteriology 114:577–591
    [Google Scholar]
  11. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 227:680–685
    [Google Scholar]
  12. McCauley J. W., Mahy B. W. J. 1983; Structure and function of the influenza virus genome. Biochemical Journal 211:281–294
    [Google Scholar]
  13. Martin S. J., Ter Meulen V. 1976; A rapid method for the quantitative study of RNA from canine distemper virus infected cells. Journal of General Virology 32:321–325
    [Google Scholar]
  14. Nelson T., Brutlag D. 1979; Addition of homopolymers to the 3′-ends of duplex DNA with terminal transferase. Methods in Enzymology 68:41–50
    [Google Scholar]
  15. Rigby P. W. J., Dieckmann M., Rhodes C., Berg P. 1977; Labelling DNA to high specific activity in vitro by nick translation with DNA polymerase I. Journal of Molecular Biology 113:237–251
    [Google Scholar]
  16. Rima B. K. 1983; The proteins of morbilliviruses. Journal of General Virology 64:1205–1219
    [Google Scholar]
  17. Roberts B. E., Patterson B. M. 1973; Efficient translation of tobacco mosaic virus RNA and rabbit globin 9S RNA in a cell-free system from commercial wheat germ. Proceedings of the National Academy of Sciences, U,. S,. A 70:2330–2334
    [Google Scholar]
  18. Roychoudhury R., Jay E., Wu R. 1976; Terminal labelling and addition of homopolymer tracts to duplex DNA fragments by terminal deoxynucleotidyl transferase. Nucleic Acids Research 3:101–116
    [Google Scholar]
  19. Rozenblatt S., Gesang C., Lavie V., Neumann F. S. 1982; Cloning and characterisation of DNA complementary to the measles virus mRNA encoding hemagglutinin and matrix proteins. Journal of Virology 42:790–797
    [Google Scholar]
  20. Stark G. R., Williams J. G. 1979; Quantitative analysis of specific labeled RNAs using DNA covalently linked to diazobenzyloxymethyl-paper. Nucleic Acids Research 6:195–203
    [Google Scholar]
  21. Ter Meulen V., Carter M. J. 1982; Morbillivirus persistent infections in animals and man. In Virus Persistence pp 97–132 Edited by Mahy B. W. J. Minson A. C., Darby G. K. Cambridge: Cambridge University Press;
    [Google Scholar]
  22. Thomas P. S. 1980; Hybridisation of denatured RNA and small DNA fragments transferred to nitrocellulose. Proceedings of the National Academy of Sciences, U. S. A 77:5201–5205
    [Google Scholar]
  23. Van Der Werf S., Bregerere F., Kopecka H., Kitamura N., Rothberg P. G., Kaurilsky P., Wimmer E., Girard M. 1981; Molecular cloning of the genome of poliovirus type 1. Proceedings of the National Academy of Sciences, U,. S,. A 78:5983–5987
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-65-3-549
Loading
/content/journal/jgv/10.1099/0022-1317-65-3-549
Loading

Data & Media loading...

Most cited Most Cited RSS feed