Molotov S V, Duzhiĭ D E, Danilevich V N, Sukhodolets V V
Mol Gen Mikrobiol Virusol. 1990 Apr(4):10-4.
The beta-galactosidase gene from the chromosome of Streptococcus thermophilus, strain 6 kb, has been cloned on a vector plasmid pBR322. The corresponding gene has been found to be located on the Pst1 DNA fragment. The restriction map of this 6 kb fragment has been constructed. The shortening of the DNA fragment carrying the beta-galactosidase gene has been achieved by digestion of the recombinant derivative of pBR322 by the restriction endonuclease Sau3A under the conditions of incomplete hydrolysis. The obtained fragments have been cloned into the BamHI site in the berepliconed shuttle vector pCB20 for grampositive and gramnegative bacteria. The obtained recombinant plasmids contained the beta-galactosidase gene in the inserted fragments of different length. Expression of the cloned beta-galactosidase gene in Escherichia coli and Bacillus subtilis cells has been studied.
嗜热链球菌6 kb染色体上的β-半乳糖苷酶基因已克隆到载体质粒pBR322上。已发现相应基因位于Pst1 DNA片段上。构建了该6 kb片段的限制性图谱。在不完全水解条件下,用限制性内切酶Sau3A消化pBR322的重组衍生物,实现了携带β-半乳糖苷酶基因的DNA片段的缩短。将获得的片段克隆到用于革兰氏阳性和革兰氏阴性细菌的双复制穿梭载体pCB20的BamHI位点。获得的重组质粒在不同长度的插入片段中含有β-半乳糖苷酶基因。研究了克隆的β-半乳糖苷酶基因在大肠杆菌和枯草芽孢杆菌细胞中的表达。