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. 1971 Sep;68(9):2203–2206. doi: 10.1073/pnas.68.9.2203

Distinguishing Reverse Transcriptase of an RNA Tumor Virus from Other Known DNA Polymerases

N C Goodman 1, S Spiegelman 1
PMCID: PMC389385  PMID: 4332248

Abstract

Assays are described that permit one to distinguish the reverse transcriptase of RNA tumor viruses from known normal cellular DNA-instructed DNA polymerases. Template responses of purified reverse transcriptase were compared with those of similar preparations of the DNA polymerase I of Escherichia coli and of calf-thymus polymerase. All three enzymes responded well to the synthetic duplexes poly(dT)·poly(A), poly(U)·poly(A), and poly(dT)·poly(dA). Hence, these duplexes can detect, but cannot distinguish reverse, transcriptase from the known normal DNA polymerases. However, certain oligomer-homopolymer complexes serve as excellent distinguishing agents. The reverse transcriptase responds very well to (dT)10·poly(A) and very poorly to (dT)10·poly(dA), whereas both cellular DNA polymerases do not exhibit this behavior.

Purified single-stranded RNA also serves as a diagnostic device, since only reverse transcriptase gives a detectable response. To be definitive, a positive response to RNA must be accompanied by a demonstration via molecular hybridization that the DNA product is complementary to the RNA and not to some minor DNA contaminant.

Keywords: calf thymus, Qβ phage, avian myeloblastosis virus

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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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