Cloning of cDNA to a yeast viral double-stranded RNA and comparison of three viral RNAs

Libuse A. Bobek, Jeremy A. Bruenn, Loren Field, Kenneth W. Gross

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

We have constructed recombinant DNA clones containing small complementary DNA (cDNA) sequences homologous to portions of a 4.8-kb yeast viral double-stranded RNA (dsRNA) (L1) that codes for the viral capsid polypeptide. Neither the viral dsRNA nor its in vitro transcript is polyadenylated; hence the cDNAs were synthesized by reverse transcriptase on the in vitro mRNA transcript made by the viral transcriptase, using sheared salmon sperm DNA as a random primer. This is the first reported cloning of cDNA homologous to a viral double-stranded RNA. This method should be of general utility for dsRNA viruses, since all have a capsid-associated transcriptase activity. The lengths of the overlapping cDNA inserts varied from 100 to 800 bp. About 40% of them mapped to the 5' end of the in vitro transcript, and these have been ordered. At least 1485 bp of this end of L1 is represented in the cloned cDNAs characterized. Using the cloned cDNAs as probes, we have shown that the L dsRNAs of two viral subtypes are similar at the transcription initiation site and dissimilar elsewhere.

Original languageEnglish (US)
Pages (from-to)225-230
Number of pages6
JournalGene
Volume19
Issue number2
DOIs
StatePublished - 1982
Externally publishedYes

Fingerprint

Double-Stranded RNA
Viral RNA
Organism Cloning
Complementary DNA
Yeasts
Capsid
DNA-Directed RNA Polymerases
Recombinant DNA
Salmon
Transcription Initiation Site
RNA Viruses
RNA-Directed DNA Polymerase
Spermatozoa
Clone Cells
Messenger RNA
Peptides
DNA
In Vitro Techniques

Keywords

  • recombinant DNA
  • Yeast killer factor
  • yeast viral transcriptase

ASJC Scopus subject areas

  • Genetics

Cite this

Cloning of cDNA to a yeast viral double-stranded RNA and comparison of three viral RNAs. / Bobek, Libuse A.; Bruenn, Jeremy A.; Field, Loren; Gross, Kenneth W.

In: Gene, Vol. 19, No. 2, 1982, p. 225-230.

Research output: Contribution to journalArticle

Bobek, Libuse A. ; Bruenn, Jeremy A. ; Field, Loren ; Gross, Kenneth W. / Cloning of cDNA to a yeast viral double-stranded RNA and comparison of three viral RNAs. In: Gene. 1982 ; Vol. 19, No. 2. pp. 225-230.
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