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Practical procedures for genetic manipulation systems for medermycin‐producing Streptomyces sp. AM‐7161
Author(s) -
Huiqun Deng,
Xiaofeng Cai,
Jianxin Peng,
Huazhu Hong,
Koji Ichinose,
Aiying Li
Publication year - 2010
Publication title -
journal of basic microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.58
H-Index - 54
eISSN - 1521-4028
pISSN - 0233-111X
DOI - 10.1002/jobm.200900240
Subject(s) - protoplast , plasmid , strain (injury) , mycelium , transformation (genetics) , streptomyces , polyketide , escherichia coli , biology , mutant , bacteria , chemistry , microbiology and biotechnology , biosynthesis , computational biology , genetics , biochemistry , gene , botany , anatomy
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. (© 2010 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)