Objective To construct a Streptomyces avermitilis strain producing only "B" components by the rnreplacement of aveD gene with a resistance cassette. Methods A DNA fragment carrying aveD gene amplified by PCR were interrupted by apt (apramycin resistance gene) inserted to a restriction site of NruI, the constructed recom-binant plasmid pID03 was transferred by conjugation via E. coli ET12567 (pUZ8002) into a wild type strain, and S. avermitilis S-2 obtained a mutant strain AvcD24. Conclusions Analysis of the metabolites of AveD24 by HPLC re-vealed that the disappearance of component "A" with still remaining of component B, as expected. But two unexpected rncompounds were also identified as being oligomycin A and 5-oxoavermectin 1a by HPLC and LC-MS.%目的 探索阿维链霉菌中aveD基因插入失活后对发酵产物组分的影响.方法采用将抗生素(安普霉素)抗性基因插入到aveD基因的方法,构建了重组质粒pID03;利用接合转移的方法将重组质粒导人到阿维链霉菌S-2(Streptomyces atwraitilis S-2)菌株中,并通过抗性标记筛选双交换的菌株.结果得到了aveD基因插入失活的菌株AveD24.该菌株不再产生4个A组分,只产生4个B组分,同时还产生2个异常的组分,经HPLC和质谱分析,初步确定异常组分D24-1为寡霉素A,另一异常组分D24-2为5-酮avermectin 1a.
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