Ringrose L, Angrand P O, Stewart A F
European Molecular Biology Laboratory, Heidelberg, Germany.
Eur J Biochem. 1997 Sep 15;248(3):903-12. doi: 10.1111/j.1432-1033.1997.00903.x.
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.
整合酶家族的位点特异性重组酶氨基酸序列相似性有限,但利用共同的反应机制重组不同的DNA靶位点。在此,我们报道了来自酵母克鲁维酵母的2μm类质粒pKWS1上编码的Kw位点特异性重组酶的特性。利用体外翻译的Kw重组酶,我们证明该蛋白能够结合并重组其假定的DNA靶位点。重组是保守的,Kw靶位点有一个7个碱基对的间隔区。我们证明Kw重组酶能够在哺乳动物细胞系中介导重组,因此,它有潜力用作异源系统中基因组操作的工具。