1887

Abstract

SUMMARY

Antisera were raised against P3 (mol. wt. 32000), the full-length translation product of alfalfa mosaic virus (AlMV) RNA 3. P3 was made by translation in wheat germ extracts, using unfractionated AlMV RNA as message, and the products were then fractionated either by centrifugation through a sucrose gradient, or by phosphocellulose chromatography, each technique being followed by SDS-PAGE. Preparations made by each method were used in sequence to immunize a rabbit. The resulting antisera reacted on immunoblots with P3 synthesized , at dilutions of about 1:10000, but did not react with the translation products of AlMV RNA 1, RNA 2 or RNA 4. Although the antisera contained antibodies against some wheat germ components, most were removed by preabsorbing the antisera with the wheat germ extract. A minor wheat germ translation product of RNA 3, P′3, also reacted with the antisera. Since the mol. wt. of P′3 is apparently 1500 higher than that of P3, it is possible that P′3 is a readthrough product of the P3 cistron. With one antiserum preparation we were able to detect P3 in extracts of tobacco plants infected with AlMV.

Keyword(s): AlMV , immunoblot , protein and translation
Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-66-8-1669
1985-08-01
2024-05-21
Loading full text...

Full text loading...

/deliver/fulltext/jgv/66/8/JV0660081669.html?itemId=/content/journal/jgv/10.1099/0022-1317-66-8-1669&mimeType=html&fmt=ahah

References

  1. Berna A., Briand J. P., Stussi-Garaud C., Godefroy-Colburn T. 1984; Immunodetection of a non-structural protein of alfalfa mosaic virus (P2) in infected tobacco plants. Annales de Virologie 135E:285–296
    [Google Scholar]
  2. Bradford M. M. 1976; A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72:248–254
    [Google Scholar]
  3. Cornelissen B. J. C., Brederode F. Th., Moormann R. J. M., Bol J. F. 1983a; Complete nucleotide sequence of alfalfa mosaic virus RNA 1. Nucleic Acids Research 11:1253–1265
    [Google Scholar]
  4. Cornelissen B. J. C., Brederode F. Th., Veeneman G. H., Van Boom J. H., Bol J. F. 1983b; Complete nucleotide sequence of alfalfa mosaic virus RNA 2. Nucleic Acids Research 11:3019–3025
    [Google Scholar]
  5. Godefroy-Colburn T., Thivent C., Pinck L. 1985; Translational discrimination between the four RNAs of alfalfa mosaic virus, a quantitative evaluation. European Journal of Biochemistry 147:541–548
    [Google Scholar]
  6. Johnson D. A., Gautsch J. W., Sportsman J. R., Elder J. H. 1984; Improved technique utilizing nonfat dry milk for analysis of proteins and nucleic acids transferred to nitrocellulose. Gene Analysis Techniques 1:3–8
    [Google Scholar]
  7. Joshi S., Neeleman L., Pleij C. W. A., Haenni A. L., Chapeville F., Bosch L., Van Vloten-Doting L. 1984; ,. Non-structural alfalfa mosaic virus RNA-coded proteins present in tobacco leaf tissue. Virology 139:231–242
    [Google Scholar]
  8. Kamer G., ARGOS P. 1984; Primary structural comparison of RNA-dependent polymerases from plant, animal and bacterial viruses. Nucleic Acids Research 12:7269–7282
    [Google Scholar]
  9. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 221:680–685
    [Google Scholar]
  10. Le Roy C., Stussi-Garaud C., Hirth L. 1977; RNA-dependent RNA polymerases in uninfected and in alfalfa mosaic virus-infected tobacco plants. Virology 82:48–62
    [Google Scholar]
  11. Marcu K., Dudock B. 1974; Characterization of a highly efficient protein synthesizing system derived from commercial wheat germ. Nucleic Acids Research 1:1385–1397
    [Google Scholar]
  12. Maxam A. M., Gilbert W. 1980; Sequencing end-labeled DN A with base-specific chemical cleavages. Methods in Enzymology 65:499–560
    [Google Scholar]
  13. Nassuth A., Bol J. F. 1983; Altered balance of the synthesis of plus and minus strand RNAs induced by RNA 1 and 2 of alfalfa mosaic virus in absence of RNA 3. Virology 124:75–85
    [Google Scholar]
  14. Pinck L., Fauquet C. 1975; Analysis of pancreatic-ribonuclease-digestion products of alfalfa mosaic virus ribonucleic acid. European Journal of Biochemistry 57:441–451
    [Google Scholar]
  15. Pinck L., Hirth L. 1972; The replicative RNA and the viral RNA synthesis rate in tobacco infected with alfalfa mosaic virus. Virology 49:413–425
    [Google Scholar]
  16. Ravelonandro M., Pinck M., Pinck L. 1984; Complete nucleotide sequence of RNA 3 from alfalfa mosaic virus, Strain S. Biochimie 66:395–402
    [Google Scholar]
  17. Thivent C. 1983 Localisation des sites d’initiation fonctionels sur les RNAs du virus de la mosaique de la luzerne ‘Third cycle’ doctorate thesis Universite Louis Pasteur, Strasbourg; France:
    [Google Scholar]
  18. Towbin H., Staehelin T., GORDON J. 1979; Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proceedings of the National Academy of Sciences, U. S. A 76:4350–4354
    [Google Scholar]
  19. van Tol R. G. L., Van Vloten-Doting L. 1981; Lack of serological relationship between the 35K nonstructural protein of alfalfa mosaic virus and the corresponding proteins of three other plant viruses with a tripartite genome. Virology 109:444–447
    [Google Scholar]
  20. van Vloten-Doting L., Jaspars E. M. J. 1972; The uncoating of alfalfa mosaic virus by its own RNA. Virology 48:699–708
    [Google Scholar]
  21. Weening C. L., Bol J. F. 1975; Viral RNA replication in extracts of alfalfa mosaic-infected Vicia faba. Virology 63:77–83
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-66-8-1669
Loading
/content/journal/jgv/10.1099/0022-1317-66-8-1669
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error