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酵母DNA拓扑异构酶II由一个单拷贝的必需基因编码。

Yeast DNA topoisomerase II is encoded by a single-copy, essential gene.

作者信息

Goto T, Wang J C

出版信息

Cell. 1984 Apr;36(4):1073-80. doi: 10.1016/0092-8674(84)90057-6.

Abstract

The gene TOP2 encoding yeast topoisomerase II has been cloned by immunological screening of a yeast genomic library constructed in the phage lambda expression vector, lambda gt11. The ends of the message encoded by the cloned DNA fragment were delimited by the Berk and Sharp procedure (S1 nuclease mapping) for the 5' end and mapping of the polyA tail portion of a cDNA fragment for the 3' end. The predicted size of the message agrees with the length of the message as determined by Northern blot hybridization analysis. The identity of the gene was confirmed by expressing the gene in E. coli from the E. coli promoter lac UV5 to give catalytically active yeast DNA topoisomerase II. Disruption of one copy of the gene in a diploid yeast creates a recessive lethal mutation, indicating that the single DNA topoisomerase II gene of yeast has an essential function.

摘要

编码酵母拓扑异构酶II的基因TOP2,是通过对构建于噬菌体λ表达载体λgt11中的酵母基因组文库进行免疫筛选而克隆得到的。克隆的DNA片段所编码信息的末端,通过伯克和夏普方法(S1核酸酶图谱分析)确定5'端,通过cDNA片段的聚腺苷酸尾部分的图谱分析确定3'端。预测的信息大小与通过Northern印迹杂交分析确定的信息长度一致。通过从大肠杆菌启动子lac UV5在大肠杆菌中表达该基因以产生具有催化活性的酵母DNA拓扑异构酶II,证实了该基因的身份。在二倍体酵母中破坏该基因的一个拷贝会产生隐性致死突变,这表明酵母的单个DNA拓扑异构酶II基因具有重要功能。

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