Measurements of DNA-Loop Formation via Cre-Mediated Recombination

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Measurements of DNA-Loop Formation via Cre-Mediated Recombination

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Title: Measurements of DNA-Loop Formation via Cre-Mediated Recombination
Author(s):
Shoura, Massa J.;
Vetcher, Alexandre;
Giovan, Stefan;
Levene, Stephen D.;
Bardai, Farah;
Bharadwaj, Anusha;
Kesinger, Matthew R.
Date Created: 2011-11-29
Format: Text
Item Type: Article
Presentation
Keywords: Cre Recombinase
DNA
DNA-loop formation
Description: Presented at the Discrete and Topological Models in Molecular Biology at the University of South Florida, March 12-14, 2012.
Abstract: The Cre recombination system has become an important tool for genetic manipulation of higher organisms and a model for site-specific DNA-recombination mechanisms employed by the λ-Int superfamily of recombinases. We report a novel quantitative approach for characterizing the probability of DNA-loop formation in solution using time-dependent ensemble FRET measurements of intra- and intermolecular Cre-recombination kinetics. Our method uses an innovative technique for incorporating multiple covalent modifications at specific sites in covalently closed DNA. Because the mechanism of Cre recombinase does not conform to a simple kinetic scheme, we employ numerical methods to extract rate constants for fundamental steps that pertain to Cre-mediated loop closure. Cre recombination does not require accessory proteins, DNA supercoiling, or particular metal-ion cofactors and is thus a highly flexible system for quantitatively analyzing DNA-loop formation in vitro and in vivo.
Alternative Title: From Enzymology to DNA Popology: Measurements of DNA-Loop Formation via Cre-Mediated Recombination
Language: English
Source: Nucleic Acids Research
Link to Related Resource: http://dx.doi.org/10.1093/nar/gks430
Persistent Link: http://hdl.handle.net/10735.1/2184
Bibliographic Citation: Shoura, M. J., A. A. Vetcher, S. M. Giovan, F. Bardai, A. Bharadwaj, M. R. Kesinger, and S. D. Levene. 2012. "Measurements of DNA-Loop Formation Via Cre-Mediated Recombination." Nucleic Acids Research 40 (15): 7452-7464.
Terms of Use: CC BY-NC 3.0 (Attribution-NonCommercial)
©2012 The Author(s).

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