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[用于在枯草芽孢杆菌细胞中表达合成基因的遗传元件的化学酶法合成]

[Chemico-enzymatic synthesis of genetic elements for expression of synthetic genes in Bacillus subtilis cells].

作者信息

Gorbunov Iu A, Daniliuk N K, Il'ichev A A, Krasnykh V N, Lomakin A I

出版信息

Bioorg Khim. 1986 May;12(5):647-54.

PMID:3089232
Abstract

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.

摘要

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。

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