1887

Abstract

SUMMARY

The anomalous viroid-like RNA associated with cadang-cadang disease of coconut palms (ccRNA-1) has been cleaved by treatment with the single-strand specific nuclease S, polyadenylated and used as a template for the oligo(dT) primed synthesis of complementary DNA (cDNA) by the avian myeloblastosis virus reverse transcriptase.

The efficiency of synthesis was low, with only 3 to 4.5 ng of cDNA synthesized from 2 µg of RNA. Most of the cDNA was in the 4S size class. A Rt value of 1 × 10 mol. s/l was obtained when this cDNA was hybridized with ccRNA-1, consistent with ccRNA-1 representing a unique species of mol. wt. approx. 100000. The maximum hybridization value obtained with ccRNA-1 was approx. 50%; the S nuclease resistance of the cDNA after self-annealing was approx. 7%. The melting behaviour of the homologous hybrids provided evidence for the specificity of base-pairing with no evidence of mismatching.

The cDNA has been shown to be a specific probe for cadang-cadang associated RNA. It has been used to demonstrate that ccRNA-1 and ccRNA-2 have common nucleotide sequences, that ccRNA-1 is uniquely associated with diseased and not healthy palms and that it has no significant homology with high mol. wt. RNA or DNA from diseased palms. The value of the cDNA as a diagnostic probe for ccRNA-1 in crude nucleic acid extracts has been demonstrated.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-43-3-649
1979-06-01
2024-04-19
Loading full text...

Full text loading...

/deliver/fulltext/jgv/43/3/JV0430030649.html?itemId=/content/journal/jgv/10.1099/0022-1317-43-3-649&mimeType=html&fmt=ahah

References

  1. Bailey J. M., Davidson N. 1976; methylmercury as a reversible denaturing agent for agarose gel electrophoresis. analytical biochemistry 70:75–85
    [Google Scholar]
  2. Gonda T. J., Symons R. H. 1978; the use of hybridization analysis with complementary DNA to determine the RNA sequence homology between strains of plant viruses: its application to several strains of cucumoviruses. virology 88:361–370
    [Google Scholar]
  3. Gould A. R., Palukaitis P., Symons R. H., Mossop D. W. 1978; characterization of a satellite RNA associated with cucumber mosaic virus. virology 84:443–455
    [Google Scholar]
  4. Gould A. R., Symons R. H. 1977; determination of the sequence homology between the four RNA species of cucumber mosaic virus by hybridization analysis with complementary DNA. nucleic acids research 4:3787–3802
    [Google Scholar]
  5. Hadidi A., Jones D. M., Gillespie D. H., Wong-Staal F., Diener T. O. 1976; Hybridization of potato spindle tuber viroid to cellular DNA of normal plants. Proceedings of the National Academy of Sciences of the United States of America 73:2453–2457
    [Google Scholar]
  6. Hadidi A., Diener T. O., Modak M. J. 1977; Synthesis of DNA transcripts of potato spindle tuber viroid. FEBS Letters 75:123–127
    [Google Scholar]
  7. Harada F., Dahlberg J. E. 1975; Specific cleavage of tRNA by nuclease S1. Nucleic Acids Research 2:865–871
    [Google Scholar]
  8. Hell A., Young D. B., Birnie G. D. 1976; Synthesis of DNAs complementary to human ribosomal RNAs polyadenylated in vitro. Biochimica et Biophysica Acta 442:37–49
    [Google Scholar]
  9. Kacian D. L., Myers J. C. 1976; Synthesis of extensive, possibly complete, DNA copies of poliovirus RNA in high yields and at high specific activities. Proceedings of the National Academy of Sciences of the United States of America 73:2191–2195
    [Google Scholar]
  10. Owens R. A. 1978; In vitro synthesis and characterization of DNA complementary to potato spindle tuber viroid. Virology 89:380–387
    [Google Scholar]
  11. Owens R. A., Diener T. O. 1977; Synthesis of RNA complementary to potato spindle tuber viroid using Qβ replicase. Virology 79:109–120
    [Google Scholar]
  12. Palukaitis P., Symons R. H. 1978; Synthesis and characterization of a complementary DNA probe for chrysanthemum stunt viroid. FEBS Letters 92:268–272
    [Google Scholar]
  13. Peacock A. C., Dinoman C. W. 1968; Molecular weight estimation and separation of ribonucleic acid by electrophoresis in agarose-acrylamide composite gels. Biochemistry 7:668–674
    [Google Scholar]
  14. Peden K. W. C., Symons R. H. 1973; Cucumber mosaic virus contains a functionally divided genome. Virology 53:487–492
    [Google Scholar]
  15. Price W. C. 1971; Cadang-cadang of coconut. Indian Phytopathology 24:425–436
    [Google Scholar]
  16. Randles J. W. 1975; Association of two ribonucleic acid species with cadang-cadang disease of coconut palm. Phytopathology 65:163–167
    [Google Scholar]
  17. Randles J. W., Hatta T. 1979; Circularity of the ribonucleic acids associated with cadang-cadang disease. Virology (in the press)
    [Google Scholar]
  18. Randles J. W., Rillo E. P., Diener T. O. 1976; The viroid like structure and cellular location of anomalous RNA associated with the cadang-cadang disease. Virology 74:128–139
    [Google Scholar]
  19. Randles J. W., Boccardo G., Retuerma M. L., Rillo E. P. 1977; Transmission of the RNA species associated with cadang-cadang of coconut palm and the insensitivity of the disease to antibiotics. Phytopathology 67:1211–1216
    [Google Scholar]
  20. Rushizky O. W., Mozejko J. H. 1977; Optimization of conditions for cleavage of tRNA at the anticodon loop by S1 nuclease. Analytical Biochemistry 77:562–566
    [Google Scholar]
  21. Staynov D. Z., Pinder J. C., Gratzer W. B. 1972; Molecular weight determination of nucleic acids by gel electrophoresis in non-aqueous solution. Nature New Biology 235:108–110
    [Google Scholar]
  22. Symons R. H. 1977; The rapid simple and improved preparation of high specific activity α-(32P) dATP and α-(32P) ATP. Nucleic Acids Research 4:4347–4355
    [Google Scholar]
  23. Taylor J. M., Illmensee R., Summers J. 1976; Efficient transcription of RNA into DNA by avian sarcoma virus polymerase. Biochimica et Biophysica Acta 442:324–330
    [Google Scholar]
  24. Vogt V. M. 1973; Purification and further properties of single-strand-specific nuclease from Aspergillus oryzae. European Journal of Biochemistry 33:192–200
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-43-3-649
Loading
/content/journal/jgv/10.1099/0022-1317-43-3-649
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