Molecular cloning and nucleotide sequence analysis of the Saccharomyces cerevisiae RAD1 gene

Molecular cloning and nucleotide sequence analysis of the Saccharomyces cerevisiae RAD1 gene
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酿酒酵母RAD1基因的分子克隆和核苷酸序列分析

DOI:
10.1128/mcb.4.10.2161-2169.1984
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发表时间:
1984
影响因子:
5.3
通讯作者:
E C Friedberg
E C Friedberg
中科院分区:
生物学2区
文献类型:
--
作者:
Elizabeth Yang;E C Friedberg

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我们筛选了一个酵母基因组文库,以补充 RAD1 基因缺陷突变体的紫外线敏感性,并分离出带有 8.9 KB 插入片段的质粒,命名为 pNF1000。该多拷贝质粒定量补充了所测试的两种 rad1 突变体的紫外线敏感性,但不影响其他 rad 突变体的紫外线耐受性。通过缺失分析确定了 pNF1000 中抗紫外线功能的位置,并将推定的 RAD1 基因的内部片段整合到 RAD1 菌株的基因组中。对几个整合体的遗传分析表明整合发生在染色体RAD1位点,证明内部片段源自RAD1基因。对 pNF1000 的 3.88 KB 区域进行了测序,结果显示存在一个长 243 个核苷酸的小开放阅读框(显然与 RAD1 无关),以及一个推测编码 RAD1 蛋白的 2,916 个核苷酸的较大开放阅读框。根据两个可能的 ATG 密码子中哪一个启动翻译,RAD1 蛋白的大小计算为 110 或 97 千道尔顿。
We have screened a yeast genomic library for complementation of the UV sensitivity of mutants defective in the RAD1 gene and isolated a plasmid designated pNF1000 with an 8.9-kilobase insert. This multicopy plasmid quantitatively complemented the UV sensitivity of two rad1 mutants tested but did not affect the UV resistance of other rad mutants. The location of the UV resistance function in pNF1000 was determined by deletion analysis, and an internal fragment of the putative RAD1 gene was integrated into the genome of a RAD1 strain. Genetic analysis of several integrants showed that integration occurred at the chromosomal RAD1 site, demonstrating that the internal fragment was derived from the RAD1 gene. A 3.88-kilobase region of pNF1000 was sequenced and showed the presence of a small open reading frame 243 nucleotides long that is apparently unrelated to RAD1, as well as a 2,916-nucleotide larger open reading frame presumed to encode RAD1 protein. Depending on which of two possible ATG codons initiates translation, the size of the RAD1 protein is calculated at 110 or 97 kilodaltons.
通过整合 DNA 转化对酵母肌动蛋白基因进行致命破坏。
DOI: 10.1126/science.7046050
发表时间: 1982
期刊: Science (New York, N.Y.)
影响因子: --
作者:
Shortle,D;Haber,JE;Botstein,D
通讯作者: Botstein,D
纯化的大肠杆菌 uvrABC 蛋白的酶学特性。
DOI: 10.1073/pnas.80.20.6157
发表时间: 1983
影响因子: 11.1
作者:
Yeung,AT;Mattes,WB;Oh,EY;Grossman,L
通讯作者: Grossman,L
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
发表时间: 1983
影响因子: 11.1
作者:
Naumovski,L;Friedberg,EC
通讯作者: Friedberg,EC