从固氮菌草螺菌属血清型草螺菌菌株Z78中克隆一个类recA基因。

Cloning of a recA-like gene from the diazotroph Herbaspirillum seropedicae strain Z78.

作者信息

Steffens M B, Rigo L U, Funayama S, Souza E M, Machado H B, Pedrosa F O

机构信息

Department of Biochemistry, Universidade Federal do Paraná, Curitiba, Brazil.

出版信息

Can J Microbiol. 1993 Dec;39(12):1096-102. doi: 10.1139/m93-166.

DOI:10.1139/m93-166

PMID:8131107

Abstract

A recombinant plasmid, pBMR5, carrying a recA-like gene of Herbaspirillum seropedicae, was isolated from a H. seropedicae genomic library by intergeneric complementation of Escherichia coli recA mutant strain HB101. Quantitative survival experiments showed that pBMR5 restored the ultraviolet radiation and methyl methanesulfonate resistances and recombinational proficiency of this strain. Hybridization studies showed that there is DNA sequence homology between the recA gene of E. coli K12 and that of H. seropedicae. Restriction sites for EcoRI, HindIII, BamHI, and Bg/II were found in the DNA insert derived from H. seropedicae in pBMR5. A Tn5 insertional mutant of pBMR5, called pBMR26.2, failed to restore recombination proficiency and methyl methanesulfonate and ultraviolet resistance to recA mutants of E. coli.

摘要

通过大肠杆菌recA突变菌株HB101的属间互补，从巴西固氮螺菌基因组文库中分离出携带巴西固氮螺菌recA样基因的重组质粒pBMR5。定量存活实验表明，pBMR5恢复了该菌株对紫外线辐射和甲磺酸甲酯的抗性以及重组能力。杂交研究表明，大肠杆菌K12的recA基因与巴西固氮螺菌的recA基因之间存在DNA序列同源性。在pBMR5中源自巴西固氮螺菌的DNA插入片段中发现了EcoRI、HindIII、BamHI和Bg/II的限制性酶切位点。pBMR5的一个Tn5插入突变体，称为pBMR26.2，未能恢复大肠杆菌recA突变体的重组能力以及对甲磺酸甲酯和紫外线的抗性。