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酵母中刺激重组的DNA序列的分离

Isolation of a DNA sequence stimulating recombination in yeast.

作者信息

Mink M

机构信息

Department of Microbiology, József Attila University, Szeged, Hungary

出版信息

Acta Microbiol Hung. 1989;36(1):61-5.

PMID:2690556
Abstract

A series of DNA sequences was rescued from the yeast Saccharomyces cerevisiae transformed by a gene library and selected for the cdc35ts+, TRP1+ phenotype. These sequences did not complement the cdc35ts mutation, and were found in various amounts and orientations in degraded plasmids. A similar phenomenon was demonstrated when the HIS3 gene was cloned into one of them: a highly deleted plasmid was rescued from complemented homozygous diploid yeast cells, in which the HIS3+/his3- character was inherited at a 2:2 ratio. These results suggest that the insert sequences rescued from the cdc35ts transformants stimulate vigorous non-reciprocal recombination events by the transfer of HIS3 gene or the TRP-ARS fragment. This event was detected in the transformation of cdc35 or his3- hosts and was followed by the re-isolation of the degraded plasmid molecules.

摘要

从用基因文库转化的酿酒酵母中拯救出一系列DNA序列,并选择具有cdc35ts +、TRP1 +表型的序列。这些序列不能互补cdc35ts突变,并且在降解的质粒中以不同的数量和方向被发现。当将HIS3基因克隆到其中之一时,也证明了类似的现象:从互补的纯合二倍体酵母细胞中拯救出一个高度缺失的质粒,其中HIS3 + / his3-性状以2:2的比例遗传。这些结果表明,从cdc35ts转化体中拯救出的插入序列通过HIS3基因或TRP-ARS片段的转移刺激了强烈的非互惠重组事件。在cdc35或his3-宿主的转化中检测到了这一事件,随后重新分离出了降解的质粒分子。

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