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RuvAB蛋白在避免大肠杆菌K-12内源性tonB基因自发形成缺失突变中的作用。

Role of the RuvAB protein in avoiding spontaneous formation of deletion mutations in the Escherichia coli K-12 endogenous tonB gene.

作者信息

Mashimo Kazumi, Nagata Yuki, Kawata Masakado, Iwasaki Hiroshi, Yamamoto Kazuo

机构信息

Department of Biomolecular Sciences, Graduate School of Life Sciences, Tohoku University, Sendai 980-8577, Japan.

出版信息

Biochem Biophys Res Commun. 2004 Oct 8;323(1):197-203. doi: 10.1016/j.bbrc.2004.08.078.

Abstract

The endogenous tonB gene of Escherichia coli was used as a target for spontaneous deletion mutations which were isolated from ruvAB-, recG-, and ruvC- cells. The rates of tonB mutation were essentially the same in ruv+, ruvAB-, recG-, and ruvC- cells. We analyzed tonB mutants by sequencing. In the ruv+, recG-, and ruvC- strains, the spectra were different from those obtained from the ruvAB- cells, where deletions dominated followed by IS insertions, base substitutions, and frameshifts, in that order. We then analyzed the tonB-trp large deletion, due to simultaneous mutations of the trp operon, and found that the frequency in ruvAB- was higher than those in ruv+, recG-, and ruvC- cells. To characterize deletion formation further, we analyzed all the tonB mutants from one colicin plate. Seven deletions were identified at five sites from the 45 tonB mutants of ruv+ cells and 24 deletions at 11 sites from the 43 tonB mutants of ruvAB- cells. Thus, the ruvAB- strain is a deletion mutator. We discuss the role of RuvAB in avoiding deletions.

摘要

大肠杆菌的内源性tonB基因被用作自发缺失突变的靶点,这些突变是从ruvAB -、recG -和ruvC -细胞中分离出来的。tonB突变率在ruv +、ruvAB -、recG -和ruvC -细胞中基本相同。我们通过测序分析了tonB突变体。在ruv +、recG -和ruvC -菌株中,突变谱与从ruvAB -细胞获得的不同,在ruvAB -细胞中,缺失占主导,其次是IS插入、碱基替换和移码,顺序依次为缺失、IS插入、碱基替换、移码。然后我们分析了由于trp操纵子同时突变导致的tonB - trp大缺失,发现ruvAB -中的频率高于ruv +、recG -和ruvC -细胞中的频率。为了进一步表征缺失形成,我们分析了来自一个大肠杆菌素平板的所有tonB突变体。在ruv +细胞的45个tonB突变体中的5个位点鉴定出7个缺失,在ruvAB -细胞的43个tonB突变体中的11个位点鉴定出24个缺失。因此,ruvAB -菌株是一种缺失突变体。我们讨论了RuvAB在避免缺失中的作用。

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