Luk'ianchuk V V, Polishchuk L V
Mikrobiol Z. 2010 May-Jun;72(3):47-51.
With the help of polymerase cycle reaction a fragment of chromosome Streptomyces coelicolor A 3(2) was amplified which contains gene SCO1206 coding subunit proteins of polyketide synthase III. Cloning of amplified DNA in the structure of shuttle vectors pAX5a and pWHM4 in Escherichia coli XL1 Blue was done. Plasmids of some selected ampicillin-resistent clones were transformed into protoplasts of plasmidless variant of Streptomyces globisporus 1912. Restrictional analysis of plasmid DNA of all investigated transformants both Escherichia coli and Streptomyces globisporus 1912 showed that 1.1 kb fragments of DNA were cloned in them. Research of the complex of metabolites, synthesized by transformants of Streptomyces globisporus 1912 revealed the presence of new substances which were not synthesized by the recipient.
借助聚合酶循环反应,扩增出天蓝色链霉菌A 3(2)染色体的一个片段,该片段包含编码聚酮合酶III亚基蛋白的SCO1206基因。将扩增的DNA克隆到大肠杆菌XL1 Blue中的穿梭载体pAX5a和pWHM4结构中。将一些选定的氨苄青霉素抗性克隆的质粒转化到无质粒的球形链霉菌1912变体的原生质体中。对所有研究的转化体(包括大肠杆菌和球形链霉菌1912)的质粒DNA进行限制性分析表明,1.1 kb的DNA片段已克隆到其中。对球形链霉菌1912转化体合成的代谢物复合物的研究揭示了存在受体未合成的新物质。