期刊
BIOTECHNOLOGY LETTERS
卷 35, 期 4, 页码 571-576出版社
SPRINGER
DOI: 10.1007/s10529-012-1107-0
关键词
Gene targeting; Homologous recombination; Ku complex; NHEJ; Yarrowia lipolytica
In Yarrowia lipolytica, targeted gene replacement occurs only with long length (1 kb) homologous flanking fragments, as this yeast preferentially uses the non-homologous end-joining mechanism (NHEJ) for DNA repair over homologous recombination (HR). To improve the frequency of HR, we identified and disrupted the KU70 and KU80 genes responsible for double strand break repair in the NHEJ pathway in Y. lipolytica. In ku70a dagger HR of URA3 marker at the ADE2 locus occurred with 43 % frequency with as little as 50 bp long flanking regions. The number of Ura(+) transformants was reduced to 1 % of the Po1d (ura3-302) wild type-like strain level, regardless of the flanking fragment length. On the contrary, even though HR was not improved in ku80a dagger, Ura(+) transformants was 60 % lower compared to the wild type.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据