Advertisement

Archives of Virology

, Volume 101, Issue 1–2, pp 137–140 | Cite as

An improved procedure for the rapid one-step-cloning of full-length viroid cDNA

  • H. Puchta
  • H. L. Sänger
Rapid Communication

Summary

The efficiency of viroid cloning can be increased by three to four orders of magnitude when the synthesis of viroid cDNA is primed in such a way that it carries identical sticky ends on both termini and when the multiion transformation is applied.

Keywords

Infectious Disease Improve Procedure Viroid cDNA 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Cress DE, Kiefer MC, Owens RA (1983) Construction of infectious potato spindle tuber viroid cDNA clones. Nucleic Acids Res 11: 6821–6835Google Scholar
  2. 2.
    Diener T (ed) (1987) The viroids. Plenum Press, New YorkGoogle Scholar
  3. 3.
    Hammond RW, Owens RA (1987) Mutational analysis of potato spindle tuber viroid complex relationships between structure and infectivity. Proc Natl Acad Sci USA 84: 3967–3971Google Scholar
  4. 4.
    Hanahan D (1983) Studies on transformation of E. coli with plasmids. J Mol Biol 166: 557–580Google Scholar
  5. 5.
    Owens RA, Diener TO (1981) Sensitive and rapid diagnosis of potato spindle tuber viroid disease by nucleic acid hybridization. Science 213: 670–672Google Scholar
  6. 6.
    Puchta H, Ramm K, Sänger HL (1988) The molecular structure of hop latent viroid (HLV), a new viroid occuring worldwide in hops. Nucleic Acids Res 16: 4197–4216Google Scholar
  7. 7.
    Puchta H, Ramm K, Sänger HL (1988) Manuscript in preparationGoogle Scholar
  8. 8.
    Sano T, Uyeda I, Shikata E, Ohno T, Okada Y (1984) Nucleotide sequence of cucumber pale fruit viroid: homology to hop stunt viroid. Nucleic Acids Res 12: 3427–3434Google Scholar
  9. 9.
    Semancik JS (ed) (1987) Viroids and viroid-like pathogens. CRC Press, Boca Raton, FLGoogle Scholar
  10. 10.
    Tabler M, Sänger HL (1984) Cloned single- and double-stranded DNA copies of potato spindle tuber viroid (PSTV) and co-inoculated subgenomic DNA fragments are infectious. EMBO J 3: 3055–3062Google Scholar
  11. 11.
    Tabler M, Sänger HL (1985) Infectivity studies on different potato spindle tuber viroid (PSTV) RNAs synthesized in vitro with the SP 6 transcription system. EMBO J 4: 2191–2199Google Scholar
  12. 12.
    Tabler M, Schnölzer M, Sänger HL (1985) Molecular cloning of potato spindle tuber viroid (PSTV) cDNA synthesized by enzymatic elongation of PSTV-specific cDNA primers: a general strategy for viroid cloning. Biosci Rep 5: 143–158Google Scholar
  13. 13.
    Tsagris M, Tabler M, Mühlbach HP, Sänger HL (1987) Linear oligomeric potato spindle tuber viroid (PSTV) RNAs are accurately processedin vitro to the monomeric circular viroid proper when incubated with a nuclear extract from healthy potato cells. EMBO J 6: 2173–2183Google Scholar
  14. 14.
    Visvader JE, Symons RH (1985) Eleven new sequence variants of citrus exocortis viroid and the correlation of sequence with pathogenicity. Nucleic Acids Res 13: 2907–2919Google Scholar
  15. 15.
    Visvader JE, Symons RH (1986) Replication of in vitro-constructed viroid mutants: location of the pathogenicity-modulating domain of citrus exocortis viroid. EMBO J 5: 2051–2055Google Scholar

Copyright information

© Springer-Verlag 1988

Authors and Affiliations

  • H. Puchta
    • 1
  • H. L. Sänger
    • 1
  1. 1.Abteilung ViroidforschungMax-Planck-Institut für BiochemieMartinsried bei MünchenFederal Republic of Germany

Personalised recommendations