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- Title
RecQ Helicase-catalyzed DNA Unwinding Detected by Fluorescence Resonance Energy Transfer.
- Authors
Xing-Dong ZHANG; Shuo-Xing DOU; Ping XIE; Peng-Ye WANG; Xu Guang XI
- Abstract
A fluorometric assay was used to study the DNA unwinding kinetics induced by Escherichia coli RecQ helicase. This assay was based on fluorescence resonance energy transfer and carried out on stopped-flow, in which DNA unwinding was monitored by fluorescence emission enhancement of fluorescein resulting from helicase-catalyzed DNA unwinding. By this method, we determined the DNA unwinding rate of RecQ at different enzyme concentrations. We also studied the dependences of DNA unwinding magnitude and rate on magnesium ion concentration. We showed that this method could be used to determine the polarity of DNA unwinding. This assay should greatly facilitate further study of the mechanism for RecQ-catalyzed DNA unwinding.
- Subjects
DNA; NUCLEIC acids; DNA helicases; ISOMERASES; ENZYMES; ENERGY transfer
- Publication
Acta Biochimica et Biophysica Sinica, 2005, Vol 37, Issue 9, p593
- ISSN
1672-9145
- Publication type
Article
- DOI
10.1111/j.1745-7270.2005.00084.x