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. (UT Dallas);
Vetcher, Alexandre (UT Dallas);
Giovan, Stefan (UT Dallas);
Levene, Stephen D. (UT Dallas);
Bardai, Farah (UT Dallas);
Bharadwaj, Anusha (UT Dallas);
Kesinger, Matthew R. (UT Dallas)
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 ?-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
Link to Related Resource: http://math.usf.edu/mathbio2012/abstract_show.aspx?record=67
Persistent Link: http://hdl.handle.net/10735.1/2184
http://dx.doi.org/10.1093/nar/gks430
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: ©2012, The Author(s). Originally published by Oxford University Press. This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/ by-nc/3.0), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. Works in Treasures @ UT Dallas are made available exclusively for educational purposes such as research or instruction. Literary rights, including copyright for published works held by the creator(s) or their heirs, or other third parties may apply. All rights are reserved unless otherwise indicated by the copyright owner(s).

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