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DNA Integration by Ty Integrase in yku70Mutant Saccharomyces cerevisiae Cells

机译:Ty整合酶在yku70突变型酿酒酵母细胞中的DNA整合

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In the present work we examined nonhomologous integration of plasmid DNA in a yku70 mutant. Ten of 14 plasmids integrated as composite elements, including Ty sequences probably originating from erroneous strand-switching and/or priming events. Three additional plasmids integrated via Ty integrase without cointegrating Ty sequences, as inferred from 5-bp target site duplication and integration site preferences. Ty integrase-mediated integration of non-Ty DNA has never been observed in wild-type cells, although purified integrase is capable of using non-Ty DNA as a substrate in vitro. Hence our data implicate yKu70 as the cellular function preventing integrase from accepting non-Ty DNA as a substrate.
机译:在目前的工作中,我们研究了在 yku70 突变体中质粒DNA的非同源整合。整合为复合元件的14个质粒中有10个,包括可能源自错误链转换和/或引发事件的Ty序列。从5 bp靶位点重复和整合位点偏好推断,通过Ty整合酶整合的三个其他质粒没有共整合的Ty序列。尽管纯化的整合酶能够在体外使用非Ty DNA作为底物,但从未在野生型细胞中观察到Ty整合酶介导的非Ty DNA整合。因此,我们的数据暗示yKu70作为阻止整合子接受非Ty DNA作为底物的细胞功能。

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