Gorbunov Iu A, Daniliuk N K, Il'ichev A A, Krasnykh V N, Lomakin A I
Bioorg Khim. 1986 May;12(5):647-54.
In order to obtain the recombinant Bacillus subtilis strain, a transcriptional-translational control unit of the alpha-amylase gene of B. amyloliquefaciens was synthesized. The oligodeoxyribonucleotides were prepared by the modified triester method in solution and by the solid-phase approach. Then these oligonucleotides were joined by DNA ligase into two fragments which were cloned in the phage M13mp9 DNA and the plasmid pBR327. A plasmid harboring the site regulating the transcription of the alpha-amylase gene may be employed as vector for cloning the promoter-containing fragments in E. coli cells.
为了获得重组枯草芽孢杆菌菌株,合成了解淀粉芽孢杆菌α-淀粉酶基因的转录-翻译控制单元。寡脱氧核糖核苷酸通过溶液中的改良三酯法和固相法制备。然后,这些寡核苷酸通过DNA连接酶连接成两个片段,克隆到噬菌体M13mp9 DNA和质粒pBR327中。携带调节α-淀粉酶基因转录位点的质粒可用作载体,用于在大肠杆菌细胞中克隆含启动子的片段。