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Biosci Biotechnol Biochem. 2010;74(2):440-2. Epub 2010 Feb 7.

Insight into the mechanism of the stabilization of moloney murine leukaemia virus reverse transcriptase by eliminating RNase H activity.

Author information

1
Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Japan.

Abstract

We explored the mechanism of the stabilization of Moloney murine leukaemia virus reverse transcriptase (MMLV RT) by eliminating RNase H activity. Without the template-primer (T/P) poly(rA)-p(dT)(15), the temperature reducing initial reverse-transcription activity by 50% over a 10-min incubation of the RNase H activity-deficient variant D524A was higher by 3.7 degrees C than that of the wild-type enzyme (WT). In the reverse transcription reaction, the K(m) values for T/P of WT and D524A were almost the same. These results suggest that elimination of RNase H activity enhanced the intrinsic thermal stability of MMLV RT rather than its affinity toward T/P.

PMID:
20139597
DOI:
10.1271/bbb.90777
[Indexed for MEDLINE]
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