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奇异变形杆菌单链DNA结合蛋白(SSB)基因的克隆、测序及对大肠杆菌ssb点突变和缺失突变的互补作用

Cloning and sequencing of the Proteus mirabilis gene for a single-stranded DNA-binding protein (SSB) and complementation of Escherichia coli ssb point and deletion mutations.

作者信息

de Vries J, Wackernagel W

机构信息

Fachbereich Biologie, Universität Oldenburg, Germany.

出版信息

Microbiology (Reading). 1994 Apr;140 ( Pt 4):889-95. doi: 10.1099/00221287-140-4-889.

Abstract

The gene of Proteus mirabilis coding for a single-stranded DNA-binding protein (SSB) was cloned in Escherichia coli from a genomic library. It restored the UV resistance and the rate of cell division of an E. coli ssb-113 mutant to the same extent as the cloned E. coli ssb+ gene did. An E. coli mutant with deleted ssb was viable with the P. mirabilis ssb+ gene provided on a single-copy-number plasmid and had the same cell division rate as with the E. coli ssb+ gene on the same vector plasmid. The recovery from UV damage of an excision repair deficient (uvrA) mutant deleted for the ssb gene was identical with the ssb+ gene from P. mirabilis or E. coli, suggesting full substitution in recombinational DNA repair of the homologous by the heterologous SSB protein. The nucleotide sequence of the gene revealed that the SSB has 81% amino acid sequence homology with the E. coli SSB and only 58-63% with various plasmid SSBs. The data provide evidence that the bacterial chromosomally coded SSBs and the plasmid encoded SSBs constitute separate groups.

摘要

奇异变形杆菌编码单链DNA结合蛋白(SSB)的基因从基因组文库中克隆到大肠杆菌中。它将大肠杆菌ssb - 113突变体的紫外线抗性和细胞分裂速率恢复到与克隆的大肠杆菌ssb⁺基因相同的程度。缺失ssb的大肠杆菌突变体在单拷贝数质粒上携带奇异变形杆菌ssb⁺基因时能够存活,并且其细胞分裂速率与在同一载体质粒上携带大肠杆菌ssb⁺基因时相同。缺失ssb基因的切除修复缺陷型(uvrA)突变体从紫外线损伤中的恢复情况与来自奇异变形杆菌或大肠杆菌的ssb⁺基因相同,这表明在重组DNA修复中异源SSB蛋白可完全替代同源蛋白。该基因的核苷酸序列显示,该SSB与大肠杆菌SSB具有81%的氨基酸序列同源性,而与各种质粒SSB的同源性仅为58 - 63%。这些数据证明细菌染色体编码的SSB和质粒编码的SSB构成不同的组。

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