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Increased homologous integration frequency in Yarrowia lipolytica strains defective in non-homologous end-joining

机译:解脂耶氏酵母菌株在非同源末端连接中缺陷的同源整合频率增加

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摘要

The ascomycetous yeast Yarrowia lipolytica has been established as model system for studies of several research topics as well as for biotechnological processes in the last two decades. However, frequency of heterologous recombination is high in this yeast species, and so knockouts of genes are laborious to achieve. Therefore, the aim of this study was to check whether a reduction of non-homologous end-joining (NHEJ) of double strand breaks (DSB) results in a strong increase of proportion of homologous recombinants. The Ku70-Ku80 heterodimer is known as an essential protein complex of the NHEJ. We show that deletion of YIKU70 and/or YIKU80 results in an increase of the rate of transformants with homologous recombination (HR) up to 85 % in each case. However, it never reaches near 100 % of HR in any case as described for some other yeast. Furthermore, we demonstrated that growth of Aylku strains was similar to that of the wild-type strain.
机译:在过去的二十年中,作为辅助系统的酵母菌解脂耶氏酵母被建立为模型系统,用于研究多个研究主题以及生物技术过程。然而,在该酵母物种中异源重组的频率很高,因此基因敲除很费力。因此,本研究的目的是检查双链断裂(DSB)的非同源末端连接(NHEJ)的减少是否导致同源重组体比例的大幅增加。 Ku70-Ku80异二聚体是NHEJ的必需蛋白复合物。我们表明,在每种情况下,YIKU70和/或YIKU80的缺失都会导致转化体与同源重组(HR)的比率增加,最高可达85%。但是,在任何情况下,它都永远不会达到HR的100%,就像其他酵母所述的那样。此外,我们证明了Aylku菌株的生长与野生型菌株相似。

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