Li Y A, Sun Y P, Shi L Y, Li X P, Li H L
Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences, Beijing.
Chin J Biotechnol. 1991;7(1):43-9.
The midecamycin producer, S. mycarofaciens mutant, which has propionyl acylase activity, can convert spiramycin into propionyl spiramycin. Plasmid pIJ702 was used as a vector for the cloning of propionyl acylase gene. After shot-gun cloning, the DNA fragments of the mutant were cloned in S. lividans TK54. The results of TLC and HPLC showed that No.9 transformant can convert spiramycin into propionyl spiramycin. It demonstrated that the propionyl acylase gene was cloned and expressed in S. lividans TK54. The insert fragment of No.9 recombinant plasmid is about 4.16 kb. Southern hybridization showed that the fragment originated from S. mycarofaciens mutant. The restriction endonuclease map of No.9 recombinant plasmid was constructed.
麦迪霉素产生菌——具有丙酰基转移酶活性的变铅青链霉菌突变株，可将螺旋霉素转化为丙酰螺旋霉素。质粒pIJ702用作丙酰基转移酶基因克隆的载体。经鸟枪法克隆后，将突变株的DNA片段克隆到变铅青链霉菌TK54中。薄层层析（TLC）和高效液相色谱（HPLC）结果表明，9号转化子可将螺旋霉素转化为丙酰螺旋霉素。这表明丙酰基转移酶基因已在变铅青链霉菌TK54中克隆并表达。9号重组质粒的插入片段约为4.16 kb。Southern杂交表明该片段源自变铅青链霉菌突变株。构建了9号重组质粒的限制性内切酶图谱。
