title: Generation of DNA circles in yeast by inducible site-specific recombination.
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.
authors.affiliation: Department of Pharmacology, Robert Wood Johnson Medical School, University of Medicine and Dentistry of New Jersey, Piscataway, NJ, USA. gartenbe@umdnj.edu
authors.forename: Marc R
authors.initials: MR
authors.lastname: Gartenberg
mesh_terms: D001483:Base Sequence; D015825:Chromosomes, Fungal / Q000378:metabolism; D004254:DNA Nucleotidyltransferases / Q000378:metabolism*; D004262:DNA Restriction Enzymes / Q000378:metabolism; D004270:DNA, Circular / Q000235:genetics* / Q000302:isolation & purification; D004271:DNA, Fungal / Q000235:genetics* / Q000302:isolation & purification; D008967:Molecular Biology / Q000379:methods*; D008969:Molecular Sequence Data; D011995:Recombination, Genetic; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D013997:Time Factors
chemical_list: D004270:DNA, Circular; D004271:DNA, Fungal; D004254:DNA Nucleotidyltransferases; C077570:Site-specific recombinase; D004262:DNA Restriction Enzymes