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RecJ和RecQ蛋白在大肠杆菌K12内源性tonB基因缺失突变自发形成中的作用。

Roles of the RecJ and RecQ proteins in spontaneous formation of deletion mutations in the Escherichia coli K12 endogenous tonB gene.

作者信息

Mashimo Kazumi, Kawata Masakado, 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

Mutagenesis. 2003 Jul;18(4):355-63. doi: 10.1093/mutage/geg004.

Abstract

The endogenous tonB gene of Escherichia coli was used as a target for spontaneous deletion mutations which were isolated from recJ(-) and recQ(-) cells. Large deletions, due to simultaneous mutations of the trp operon, were also isolated. The rates of tonB mutation were 2.77 x 10(-8), 4.13 x 10(-8) and 5.00 x 10(-8) for rec(+), recJ(-) and recQ(-) cells, respectively. We analyzed 94 and 99 tonB mutants from the recJ(-) and recQ(-) cells, respectively, by sequencing. We found that IS insertion dominated, followed by base substitutions, frameshifts and deletions in both recJ(-) and recQ(-) strains. We then analyzed 55 tonB-trp deletions, ranging in size from 5907 to 20,832 bp, from the recJ(-) strains and 47 tonB-trp deletions, ranging in size from 4,959 to 16,390 bp from the recQ(-) strains. About one-third of tonB-trp deletions from both the recJ(-) and the recQ(-) cells were found to have occurred between short sequence repeats at the deletion termini. About one-third of tonB-trp deletions from both mutants showed 2-4 bp repeats in the immediate vicinity of the endpoints, which appeared to indicate no clear association with deletion. The remaining one-third of tonB-trp deletions had no homology at the endpoint. These results were similar to those for the rec(+) cells. Hanada and colleagues demonstrated that structually similar rearrangements arising during lambda bio phage formation (illegitimate recombination) increased in the recQ(-) strain. To explain this discrepancy, we interpreted as distinctive the mechanism for rearrangement during transducing phage formation which is recQ-dependent and that for deletions formed in chromosomes which is recQ-independent.

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recJ(-)和recQ(-)细胞中分离出来的。同时,由于色氨酸操纵子的同时突变,也分离出了大的缺失突变。rec(+)、recJ(-)和recQ(-)细胞的tonB突变率分别为2.77×10(-8)、4.13×10(-8)和5.00×10(-8)。我们分别对recJ(-)和recQ(-)细胞中的94个和99个tonB突变体进行了测序分析。我们发现,在recJ(-)和recQ(-)菌株中,IS插入占主导地位,其次是碱基替换、移码和缺失。然后,我们分析了recJ(-)菌株中55个大小从5907到20832 bp的tonB-trp缺失以及recQ(-)菌株中47个大小从4959到16390 bp的tonB-trp缺失。发现recJ(-)和recQ(-)细胞中约三分之一的tonB-trp缺失发生在缺失末端的短序列重复之间。两个突变体中约三分之一的tonB-trp缺失在端点附近显示出2-4 bp的重复,这似乎表明与缺失没有明显关联。其余三分之一的tonB-trp缺失在端点处没有同源性。这些结果与rec(+)细胞的结果相似。花田及其同事证明,在λ生物噬菌体形成过程中出现的结构相似的重排(非法重组)在recQ(-)菌株中增加。为了解释这种差异,我们将转导噬菌体形成过程中依赖recQ的重排机制与染色体中形成的不依赖recQ的缺失机制解释为不同的机制。

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