四环素产生菌质粒DNA的分离检测及遗传学分析

ISOLATION, IDENTIFICATION AND GENETIC ANALYSIS OF PLASMID DNA FROM A PRODVCER OF TETRACYCLINE

用７％蔗糖的ＹＥＭＥ液体培养基培养四环素产生菌金色链霉菌（Ｓｔｒｅｐｔｏｍｙｃｅｓａｕｒｅｏｆａｃｉｅｎｓ）９１－０６，以链霉菌遗传操作法［８］为基础，从该菌体中分离到质粒ｐＳＡ３１４，分子量约为２８．５ｋｂ（１８．８ＭＤ）．该质粒经酶切分析表明，具有２个ＢａｍＨＩ位点和１个ＥｃｏＲＩ位点。用 啶橙处理获得无质粒株，其在孢子形成和菌落形态方面出现明显差异。

By means of alkaline lysis method of T. Kieser, plasmid pSA314 was isolaied from the producer of tetracycline, Streptomyces aureofaciens 91-06, which grew in YEME liquid media containing 7% sucrose. Its molecuar weight is about 28. 5kb (18x106 daltons). The plasmid DNA was digested into two fragments with BamHI and one fragment with EooRI.There are differences in spore formation and clone morphology between plasmid-harboring and plasmid-cured strains which have been isolated after treating the original strains with acridine orange.