Open AccessSite-specific cleavage of chromosomesin vitrothrough Cre-loxrecombination
Author(s) -
Min Qin,
Elsa Lee,
Todd R. Zankel,
David W. Ow
Publication year - 1995
Publication title -
nucleic acids research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 9.008
H-Index - 537
eISSN - 1362-4954
pISSN - 0305-1048
DOI - 10.1093/nar/23.11.1923
Subject(s) - biology , schizosaccharomyces pombe , flp frt recombination , genetics , chromosome , homologous recombination , cre lox recombination , chromosome engineering , eukaryotic chromosome fine structure , genome , recombination , oligonucleotide , genetic recombination , microbiology and biotechnology , gene , saccharomyces cerevisiae , transgene , genetically modified mouse
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.