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

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

Research output: Contribution to journalArticle

11 Scopus citations

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 - Sep 1982
Externally publishedYes

Keywords

  • Yeast killer factor
  • recombinant DNA
  • yeast viral transcriptase

ASJC Scopus subject areas

  • Genetics

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