A DNA repair gene required for the incision of damaged DNA is essential for viability in Saccharomyces cerevisiae.

A DNA repair gene required for the incision of damaged DNA is essential for viability in Saccharomyces cerevisiae.
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切割受损 DNA 所需的 DNA 修复基因对于酿酒酵母的生存至关重要。

DOI:
10.1073/pnas.80.15.4818
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发表时间:
1983
影响因子:
11.1
通讯作者:
Friedberg,EC
Friedberg,EC
中科院分区:
综合性期刊1区
文献类型:
--
作者:
Naumovski,L;Friedberg,EC

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用含有克隆的酵母RAD 3基因的内部片段的整合质粒转化酿酒酵母的二倍体菌株(RAD 3/RAD 3)。通过同源重组的整合使二倍体RAD 3基因之一失活,产生隐性突变。这种突变被推断为在单倍体细胞中是致死的,因为二倍体转化体的孢子形成在每个四分体中分离出两个活的和两个不活的孢子,而含有RAD 3基因的一端或另一端的质粒的整合导致正常分离的二倍体转化体--即,每个四分体中有四个活孢子。DNA提供的证据表明,整合的内部片段具体发生在一个RAD 3基因的二倍体。DNA杂交。此外,转化RAD 3基因杂合的二倍体菌株(RAD 3/rad 3 -2)(携带不影响单倍体细胞活力的rad 3突变)导致一半转化体的rad-表型,表明RAD 3基因在这些细胞中失活。
A diploid strain (RAD3/RAD3) of Saccharomyces cerevisiae was transformed with an integration plasmid containing an internal fragment of the cloned yeast RAD3 gene. Integration by homologous recombination inactivated one of the diploid RAD3 genes, creating a recessive mutation. This mutation is inferred to be lethal in haploid cells since sporulation of diploid transformants segregated two viable and two inviable spores per tetrad, while integration of plasmids containing one or the other end of the RAD3 gene resulted in diploid transformants that segregated normally--i.e., four viable spores in each tetrad. Evidence that integration of the internal fragment occurred specifically at one of the RAD3 genes in the diploid is provided by DNA . DNA hybridizations. In addition, transformation of a diploid strain heterozygous for the RAD3 gene (RAD3/rad3-2) (carrying a rad3 mutation that does not affect the viability of haploid cells) results in the rad- phenotype in half of the transformants, indicating that the RAD3 gene was inactivated in these cells.
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