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[编码主要冷休克蛋白的cspA基因插入突变对大肠杆菌辐射抗性的影响]

[Effect of insertional mutation in the cspA gene encoding the major cold-shock protein on radiation resistance of Escherichia coli].

作者信息

Verbenko V N, Kuznetsova L V, Smol'nikova A V, Kalinin V L

机构信息

Petersburg Konstantinov Institute of Nuclear Physics, Russian Academy of Sciences, Gatchina, 188300 Russia.

出版信息

Genetika. 2003 Jun;39(6):748-57.

Abstract

Plasmid pCspA::Km carrying a cloned mutant allele of the cspA gene for the major Escherichia coli cold-shock protein CspA with an insertion of the kanamycin resistance gene cassette from transposon Tn903 into the core region of the coding sequence causes a 2.3-fold increase in radioresistance of wild-type E. coli cells (cspA+). The radioprotective effect of this plasmid is abolished or drastically reduced in mutants recA13 and rpoH15 defective in RecA protein and in induction of the heat-shock protein regulon, respectively. Plasmid pCspA::Km causes a 1.3-fold elevation in the resistance to gamma-irradiation of E. coli mutants with an intermediate level of radioresistance (Gamr445 and KS0160) but slightly diminishes resistance of a highly radiation-resistant Gamr445 mutant. In the chromosome of E. coli with normal DNA repair systems, the cspA::Km mutation in the homozygous state enhances resistance to the lethal effect of gamma-rays and UV light 2.9 and 1.4 times, respectively. These data suggest that the system of cold-shock proteins can modulate resistance of E. coli cells to the lethal effect of gamma-rays and UV light.

摘要

携带克隆的大肠杆菌主要冷休克蛋白CspA的cspA基因突变等位基因的质粒pCspA::Km,其编码序列的核心区域插入了来自转座子Tn903的卡那霉素抗性基因盒,导致野生型大肠杆菌细胞(cspA+)的抗辐射能力提高了2.3倍。分别在RecA蛋白缺陷的recA13突变体和热休克蛋白调控子诱导缺陷的rpoH15突变体中,该质粒的辐射防护作用被消除或大幅降低。质粒pCspA::Km使具有中等抗辐射水平的大肠杆菌突变体(Gamr445和KS0160)对γ射线的抗性提高了1.3倍,但略微降低了高度抗辐射的Gamr445突变体的抗性。在具有正常DNA修复系统的大肠杆菌染色体中,纯合状态的cspA::Km突变分别使对γ射线和紫外线致死效应的抗性提高了2.9倍和1.4倍。这些数据表明,冷休克蛋白系统可以调节大肠杆菌细胞对γ射线和紫外线致死效应的抗性。

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