The Kw recombinase, an integrase from Kluyveromyces waltii.
Site-specific recombinases of the integrase family share limited amino-acid-sequence similarity, but use a common reaction mechanism to recombine distinct DNA target sites. Here we report the characterisation of the Kw site-specific recombinase, encoded on the 2 mu-like plasmid pKWS1 from the yeast Kluyveromyces waltii. Using in vitro-translated Kw recombinase, we show that the protein is able to bind and to recombine its putative DNA target site. Recombination is conservative and the Kw target site has a spacer of seven base pairs. We show that Kw recombinase is able to mediate recombination in a mammalian cell line, thus, it has potential for use as a tool for genomic manipulation in heterologous systems.
Ringrose
L
L
European Molecular Biology Laboratory, Heidelberg, Germany.
Angrand
P O
PO
Stewart
A F
AF
D002460:Cell Line; D004247:DNA / Q000235:genetics / Q000378:metabolism*; D004254:DNA Nucleotidyltransferases / Q000235:genetics / Q000378:metabolism*; D017930:Genes, Reporter; D019426:Integrases / Q000235:genetics / Q000378:metabolism*; D007716:Kluyveromyces / Q000201:enzymology*; D007763:Lac Operon; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D010957:Plasmids / Q000235:genetics; D045522:Recombinases; D011995:Recombination, Genetic; D015183:Restriction Mapping; D017422:Sequence Analysis, DNA; D014162:Transfection
D009838:Oligodeoxyribonucleotides; D045522:Recombinases; D004247:DNA; D004254:DNA Nucleotidyltransferases; D019426:Integrases; C115006:integron integrase IntI1