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. 1974 Aug;71(8):2951–2955. doi: 10.1073/pnas.71.8.2951

Isolation of DNA Strand-Specific Early Messenger RNA Species in Cells Infected by Human Adenovirus 2

Werner Büttner 1, Zsuzsanna Veres-Molnár 1, Maurice Green 1
PMCID: PMC388596  PMID: 4528619

Abstract

Hybridization to the separated light (L) and heavy (H) strands of adenovirus 2 DNA in 50% formamide at 37° was used to isolate undegraded virus-specific RNA molecules from the polyribosomes of cycloheximide-treated human KB cells early after infection with adenovirus 2. About 20% of polyribosomal RNA labeled with [3H]uridine from 4 to 7 hr after infection was virus-specific. Twice as much labeled RNA was homologous to the L strand as to the H strand. Polyacrylamide gel electrophoresis of RNA selected with unfractionated adenovirus DNA resolved a major component of virus-specific RNA in the 19-20 S region of the gel and smaller amounts of viral RNA in two heterogeneous fractions migrating at 15-18 S and 21-26 S. Selection with individual DNA strands showed that the 19-20 S main size class of early mRNA consists of two homogeneous RNA species with slightly different mobilities, the transcripts from the L and H strand having molecular weights of 7.4 × 105 and 7.7 × 105, respectively. The 15-18 S RNA hybridized with the L strand and the 21-26 S RNA with the H strand.

Keywords: DNA·RNA hybridization, gel electrophoresis, cycloheximide

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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