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Homologous recombination between single-stranded DNA and chromosomal genes in Saccharomyces cerevisiae.

机译:酿酒酵母中单链DNA和染色体基因之间的同源重组。

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摘要

Transformation of Saccharomyces cerevisiae strains was examined by using the URA3 and TRP1 genes cloned into M13 vectors in the absence of sequences capable of promoting autonomous replication. These constructs transform S. cerevisiae cells to prototrophy by homologous recombination with the resident mutant gene. Single-stranded DNA was found to transform S. cerevisiae cells at efficiencies greater than that of double-stranded DNA. No conversion of single-stranded transforming DNA into duplex forms could be detected during the transformation process, and we conclude that single-stranded DNA may participate directly in recombination with chromosomal sequences. Transformation with single-stranded DNA gave rise to both gene conversion and reciprocal exchange events. Cotransformation with competing heterologous single-stranded DNA specifically inhibited transformation by single-stranded DNA, suggesting that one of the components in the transformation-recombination process has a preferential affinity for single-stranded DNA.
机译:在不存在能够促进自主复制的序列的情况下,通过使用克隆到M13载体中的URA3和TRP1基因来检查酿酒酵母菌株的转化。这些构建体通过与驻留突变基因的同源重组将酿酒酵母细胞转化为原生质。发现单链DNA以比双链DNA更高的效率转化酿酒酵母细胞。在转化过程中,未检测到单链转化DNA转化为双链体形式,我们得出的结论是,单链DNA可能直接参与染色体序列的重组。用单链DNA转化既引起基因转化又发生相互交换事件。与竞争性异源单链DNA的共转化可特异性抑制单链DNA的转化,这表明转化重组过程中的一种组分对单链DNA具有优先的亲和力。

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