Fernández de Henestrosa A R, Barbé J
Department of Genetics and Microbiology, Faculty of Sciences, Autonomous University of Barcelona, Bellaterra, Spain.
Mutat Res. 1994 Jul 1;308(1):99-107. doi: 10.1016/0027-5107(94)90202-x.
A fusion between the recA gene of Rhizobium phaseoli and the lacZ gene was constructed in vitro and cloned in a mini-Tn5 transposon derivative to obtain chromosomal insertions which make it possible to quantitatively examine their transcriptional regulation in both R. phaseoli and E. coli. Likewise, and by insertion of a spectinomycin-resistance gene cassette into the recA gene of R. phaseoli and subsequent marker exchange, a RecA- derivative of this bacterial species has been obtained. Analysis of this recA-lacZ fusion showed that it was inducible by DNA damage in the RecA+ strain of R. phaseoli but not in the RecA- mutant. On the other hand, the recA-lacZ fusion of R. phaseoli was not induced in DNA-damaged RecA+ cells of E. coli. Furthermore, the range of UV doses which give rise to dose dependence in the induction of its respective recA genes is different in R. phaseoli from that in E. coli.
菜豆根瘤菌recA基因与lacZ基因的融合体在体外构建,并克隆到mini-Tn5转座子衍生物中,以获得染色体插入片段,从而能够定量检测它们在菜豆根瘤菌和大肠杆菌中的转录调控。同样,通过将壮观霉素抗性基因盒插入菜豆根瘤菌的recA基因并随后进行标记交换,获得了该细菌物种的RecA-衍生物。对这种recA-lacZ融合体的分析表明,它在菜豆根瘤菌的RecA+菌株中可被DNA损伤诱导,但在RecA-突变体中不能被诱导。另一方面,菜豆根瘤菌的recA-lacZ融合体在DNA损伤的大肠杆菌RecA+细胞中未被诱导。此外,在菜豆根瘤菌中,引起其各自recA基因诱导剂量依赖性的紫外线剂量范围与大肠杆菌中的不同。