Single molecule kinetics of reverse transcriptase

Charles M Schroeder, Sangjin Kim, Paul C. Blainey, X. Sunney Xie

Research output: Contribution to conferencePaper

Abstract

We use single molecule methods to directly observe the enzymatic behavior of individual reverse transcriptase (RT) molecules on single-stranded DNA template strands in vitro. A flow-stretched DNA assay is used, allowing for characterization of enzymatic rate, processivity, and pausing of RT during DNA synthesis as a function of template base pair content, secondary structure, and applied template tension. We study the kinetics of plus-strand DNA synthesis catalyzed by RT derived from the Moloney murine leukemia virus (M-MLV) with inhibited RNase H activity, an enzyme commonly used for RT-PCR. Initial observations show an average enzymatic rate of ∼5-10 bp/sec. Understanding potentially diverse kinetic mechanisms in molecular subpopulations may allow for development of more effective treatments for retroviral infections leading to cancer.

Original languageEnglish (US)
Number of pages1
StatePublished - Dec 1 2005
Externally publishedYes
Event05AIChE: 2005 AIChE Annual Meeting and Fall Showcase - Cincinnati, OH, United States
Duration: Oct 30 2005Nov 4 2005

Other

Other05AIChE: 2005 AIChE Annual Meeting and Fall Showcase
CountryUnited States
CityCincinnati, OH
Period10/30/0511/4/05

ASJC Scopus subject areas

  • Engineering(all)

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    Schroeder, C. M., Kim, S., Blainey, P. C., & Xie, X. S. (2005). Single molecule kinetics of reverse transcriptase. Paper presented at 05AIChE: 2005 AIChE Annual Meeting and Fall Showcase, Cincinnati, OH, United States.