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Chromatin Remodeling

Volume 833 of the series Methods in Molecular Biology pp 103-113

Date:

Generation of DNA Circles in Yeast by Inducible Site-Specific Recombination

  • Marc R. GartenbergAffiliated withDepartment of Pharmacology, Robert Wood Johnson Medical School, University of Medicine and Dentistry of New JerseyThe Cancer Institute of New Jersey Email author 

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Abstract

Site-specific recombinases have been harnessed for a variety of genetic manipulations involving the gain, loss, or rearrangement of genomic DNA in a variety of organisms. The enzymes have been further exploited in the model eukaryote Saccharomyces cerevisiae for mechanistic studies involving chromosomal context. In these cases, a chromosomal element of interest is converted into a DNA circle within living cells, thereby uncoupling the element from neighboring regulatory sequences, obligatory chromosomal events, and other context-dependent effects that could alter or mask intrinsic functions of the element. In this chapter, I discuss general considerations in using site-specific recombination to create DNA circles in yeast and the specific application of the R recombinase.

Key words

Site-specific recombination R Recombinase RS site DNA circle Saccharomyces cerevisiae