The College of Life Sciences, Central China Normal University, Wuhan 430079, China.
J Basic Microbiol. 2010 Jun;50(3):299-301. doi: 10.1002/jobm.200900240.
Streptomyces sp. AM-7161 is a producer of an aromatic polyketide medermycin with strong antibacterial and antitumor activity. It has been inefficient to perform genetic manipulations in this strain, which heavily hinders the genetic analysis of some interesting problems concerning tailoring modifications in medermycin biosynthesis. Here, condition optimizations of sporulation and mycelium growth of this strain as well as protoplast preparation and regeneration were conducted systematically. Based on these results, protoplast transformation system was established, and two types of foreign plasmids (integrative and auto-replicating) were efficiently introduced into this strain. Additionally, a conjugation system to mediate plasmid transfer between Escherichia coli and Streptomyces sp. AM-7161 was also optimized and established. These results provide the practical procedures to perform more convenient genetic analysis of medermycin biosynthesis in this strain.
链霉菌 AM-7161 是一种产生芳香聚酮类美替麦辛的微生物，具有很强的抗菌和抗肿瘤活性。该菌株的遗传操作效率较低，严重阻碍了对美替麦辛生物合成中一些有趣的修饰问题的遗传分析。在这里，我们系统地优化了该菌株的孢子形成和菌丝生长条件，以及原生质体制备和再生条件。在此基础上，建立了原生质体转化系统，有效地将两种类型的外源质粒（整合型和自主复制型）导入该菌株。此外，还优化并建立了大肠杆菌和链霉菌 AM-7161 之间质粒转移的接合系统。这些结果为该菌株中美替麦辛生物合成的更方便的遗传分析提供了实用程序。
