Deletion analysis of oligomycin PKS genes （olmA） in Streptomyces arermitilis
Deletion analysis of oligomycin PKS genes （olmA） in Streptomyces arermitilis
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发表时间：
2004
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通讯作者：
ZhangXiaolin;ChenZhi;ZHAOJinlei;SONGYuan;Wenying;LIJilun
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作者：
ZhangXiaolin;ChenZhi;ZHAOJinlei;SONGYuan;Wenying;LIJilun
利用基因缺失载体pXL 05（pKC 1139：：△ olmA 1 +△ olmA 4）破坏驱虫阿维菌素B和细胞生长抑制剂寡霉素产生菌阿维链霉菌CZ 8 -73中的寡霉素PKS编码基因（olmA），染色体上的olmA基因簇通过双交换被质粒上的缺失等位基因取代。其中4种干扰物经Southern杂交证实。摇瓶试验和HPLC分析表明，4株突变株不再产生毒性的寡霉素，而只产生4种阿维菌素组分B，即阿维菌素Bla、B1 B、B2 a、B2 B。这些突变体中阿维菌素B的产量分别与CZ 8 -73中的产量相当。这说明olmA基因的缺失并不影响阿维菌素的生物合成。缺失突变株遗传稳定，有望成为阿维菌素B工业化生产的优良菌株。
Gene deletion vector pXL05(pKC1139::△olmA1+△olmA4) was used to disrupt oligomycin PKS encoding genes (olmA ) in Streptomyces avermitilis CZ8-73, the producer of anthelmintic avermectins B and the cell growth inhibitor oligomycin, olmA gene cluster in the chromosome was displaced by deletion allele on the plasmid via double crossover. Four of disruptants were confirmed by Southern blotting. Shaking flask experiments and HPLC analyses showed that the four mutants no longer produced the toxic oligomycin, but only made four components of avermectins B, which were avermectin Bla, Blb, B2a, B2b. The yields of avermectins B in these mutants were separately equal to those in CZ8-73. This revealed that olmA genes deletion did not affect the biosynthesis of avermectins. The deletion mutants were proved to be genetically stable, and thus might be promising strains in industrial production of avermectins B.