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酿酒酵母光复活基因PHR1的克隆与定位

Cloning and mapping of Saccharomyces cerevisiae photoreactivation gene PHR1.

作者信息

Schild D, Johnston J, Chang C, Mortimer R K

出版信息

Mol Cell Biol. 1984 Sep;4(9):1864-70. doi: 10.1128/mcb.4.9.1864-1870.1984.

Abstract

The yeast Saccharomyces cerevisiae, like most organisms, is able to directly repair pyrimidine dimers by using a photoreactivating enzyme and visible light. Cells carrying the phr1 mutation were shown previously to be unable to photoreactivate dimers, but neither the map position nor the primary gene product of the PHR1 gene has been determined. We have cloned this gene and determined its map position. A plasmid containing a 6.4-kilobase yeast DNA insert has been isolated and shown to restore photoreactivation in a phr1 strain. A 3.1-kilobase subclone has also been shown to complement phr1. The original plasmid was targeted to integrate into chromosomal DNA at a site homologous to the insert by cutting within the insert. Two of these integrants have been mapped on the right arm of chromosome XV; the integrants have been further mapped at ca. 13 centimorgans from prt1. It has also been independently determined that phr1 maps at this location. Thus, we have determined the map position of PHR1 and also have shown that the plasmid contains PHR1 rather than a suppressor of the phr1 mutation.

摘要

与大多数生物体一样,酿酒酵母能够通过光复活酶和可见光直接修复嘧啶二聚体。先前已证明携带phr1突变的细胞无法使二聚体进行光复活,但PHR1基因的图谱位置和主要基因产物均未确定。我们已经克隆了该基因并确定了其图谱位置。一个含有6.4千碱基酵母DNA插入片段的质粒已被分离出来,并显示可恢复phr1菌株的光复活能力。一个3.1千碱基的亚克隆也已显示可互补phr1。通过在插入片段内切割,将原始质粒靶向整合到与插入片段同源的染色体DNA位点。其中两个整合体已定位在第十五号染色体的右臂上;这些整合体已进一步定位在距prt1约13厘摩处。独立研究也已确定phr1位于该位置。因此,我们确定了PHR1的图谱位置,并且还表明该质粒包含PHR1而非phr1突变的抑制子。

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