title: Site-specific cleavage of chromosomes in vitro through Cre-lox recombination.
abstract: Site-specific recombination systems are useful tools for chromosome engineering in vivo and site-specific DNA cleavage methods have applications in genome analysis and gene isolation. Here, we report a new method to fragment chromosomes in vitro using the Cre-lox site-specific recombination system. Two lox sites were targeted into the 5.7 Mb chromosomes I of Schizosaccharomyces pombe. In vitro recombination between chromosomal lox sites and exogenously provided lox oligonucleotides 'cleaved' the chromosome at the defined lox sequences. Site-specific cleavage of lox sites in the tobacco genome was also demonstrated. This recombination-based cleavage method provides a novel approach for structural and functional analyses of eukaryotic chromosomes as it allows direct isolation of chromosome regions that correspond to phenotypes revealed through Cre-lox mediated chromosome rearrangements in vivo. Moreover, recombination with end-labeled lox oligonucleotides would permit the specific end-labeling of chromosome segments to facilitate the long range mapping of chromosomes.
authors.affiliation: Plant Gene Expression Center, US Department of Agriculture, Albany, CA 94710, USA.
authors.forename: M
authors.initials: M
authors.lastname: Qin
authors.forename: E
authors.initials: E
authors.lastname: Lee
authors.forename: T
authors.initials: T
authors.lastname: Zankel
authors.forename: D W
authors.initials: DW
authors.lastname: Ow
mesh_terms: D001483:Base Sequence; D002875:Chromosomes / Q000235:genetics*; D008969:Molecular Sequence Data; D010957:Plasmids; D011995:Recombination, Genetic; D012568:Schizosaccharomyces / Q000235:genetics*