Grkovic S, Mahanty H K
Department of Plant and Microbial Sciences, University of Canterbury, Christchurch, New Zealand.
Mol Gen Genet. 1996 Feb 25;250(3):323-8. doi: 10.1007/BF02174390.
A recA clone was isolated from a cosmid library of Serratia entomophila constructed in the Escherichia coli strain HB101. Subcloning and transposon mutagenesis were used to identify a 1.36 kb fragment containing the recA gene. A cloned recA mutation, generated by transposon mutagenesis and the replacement of a portion of the recA gene with an antibiotic resistance cassette, was introduced into the chromosome via a marker exchange technique. The recA strains created were deficient in DNA repair, homologous recombination and both the spontaneous and UV induction of prophages. S. entomophila recA strains showed continued pathogenicity towards the New Zealand grass grub, Costelytra zealandica. Simple procedures for further construction of S. entomophila recA strains have been demonstrated.
从构建于大肠杆菌HB101菌株中的嗜虫沙雷氏菌黏粒文库中分离出一个recA克隆。通过亚克隆和转座子诱变来鉴定包含recA基因的1.36 kb片段。由转座子诱变产生并用抗生素抗性盒替换部分recA基因而形成的克隆recA突变体,通过标记交换技术导入染色体。所构建的recA菌株在DNA修复、同源重组以及原噬菌体的自发诱导和紫外线诱导方面存在缺陷。嗜虫沙雷氏菌recA菌株对新西兰牧草金龟子Costelytra zealandica仍具有致病性。已证明了进一步构建嗜虫沙雷氏菌recA菌株的简单程序。