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. 1988 Jan 11;16(1):265–277. doi: 10.1093/nar/16.1.265

Isolation of cloned Moloney murine leukemia virus reverse transcriptase lacking ribonuclease H activity.

M L Kotewicz 1, C M Sampson 1, J M D'Alessio 1, G F Gerard 1
PMCID: PMC334625  PMID: 2448747

Abstract

Retroviral reverse transcriptase possesses DNA polymerase and ribonuclease H (RNase H) activity within a single polypeptide. Chemical or proteolytic treatment of reverse transcriptase has been used in the past to produce enzyme that is missing DNA polymerase activity and retains RNase H activity. It has not been possible to obtain reverse transcriptase that lacks RNase H but retains DNA polymerase activity. We have constructed a novel deletion derivative of the cloned Moloney murine leukemia virus (M-MLV) reverse transcriptase gene, expressed the gene in E. coli, and purified the protein to near homogeneity. The purified enzyme has a fully active DNA polymerase, but has no detectable RNase H activity. These results are consistent with, but do not prove, the conclusion that the DNA polymerase and RNase H activities of M-MLV reverse transcriptase reside within separate structural domains.

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Selected References

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