Practical procedures for genetic manipulation systems for medermycin-producing Streptomyces sp AM-7161
Practical procedures for genetic manipulation systems for medermycin-producing Streptomyces sp AM-7161
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DOI：
10.1002/jobm.200900240
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发表时间：
2010-06-01
影响因子：
3.1
通讯作者：
Li Aiying
中科院分区：
文献类型：
--
作者：
Deng Huiqun;Cai Xiaofeng;Li Aiying
链霉菌AM-7161是一种芳香族聚酮美得霉素的产生菌，具有很强的抗菌和抗肿瘤活性。在该菌株中进行遗传操作的效率很低，这严重阻碍了有关麦迪霉素生物合成中剪裁修饰的一些有趣问题的遗传分析。本文对该菌株产孢条件、菌丝生长条件、原生质体制备和再生条件进行了系统的研究。在此基础上，建立了原生质体转化体系，并成功地将整合型和自复制型两种外源质粒导入该菌株。此外，还优化并建立了介导大肠杆菌与链霉菌AM-7161之间质粒转移的接合系统。这些结果提供了实用的程序，进行更方便的遗传分析，在该菌株的美得霉素生物合成。
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 con 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.