Paul K, Ghosh S K, Das J
Mol Gen Genet. 1986 Apr;203(1):58-63. doi: 10.1007/BF00330384.
A library containing more than 80% of the Vibrio cholerae genome was constructed by cloning BamH1 restriction fragments into pBR322. Using interspecific complementation of an Escherichia coli recA mutant with plasmids containing the gene bank of V. cholerae, a recA-like gene was identified. The recombinant plasmid, designated as pDP145, contained a 1.45 kb segment of V. cholerae DNA which codes for a protein of molecular weight 39,000. The product of this gene confers methyl methane sulphonate resistance on the E. coli recA mutant, suppresses its ultraviolet (UV) light sensitive phenotype and has proteolytic activity on the phage lambda repressor. Induction of a 39,000 dalton protein in UV-irradiated V. cholerae cells was demonstrated.
通过将霍乱弧菌的BamH1限制性片段克隆到pBR322中构建了一个包含80%以上霍乱弧菌基因组的文库。利用大肠杆菌recA突变体与含有霍乱弧菌基因文库的质粒进行种间互补,鉴定出一个recA样基因。重组质粒命名为pDP145,含有一段1.45kb的霍乱弧菌DNA片段,该片段编码一种分子量为39000的蛋白质。该基因的产物赋予大肠杆菌recA突变体对甲基磺酸甲酯的抗性,抑制其紫外线(UV)敏感表型,并对噬菌体λ阻遏物具有蛋白水解活性。证明了紫外线照射的霍乱弧菌细胞中诱导出一种39000道尔顿的蛋白质。