阿维链霉菌中aveD基因插入失活产生的异常组分(英文)  

Unusual metabolites produced by recombinant Streptomyces avermitilis after insertional inactivation into aveD gene

作　　者：陈红霞[1] 何建勇[1] 张怡轩[1] 

机构地区：[1]沈阳药科大学生命科学与生物制药学院,沈阳110016

出　　处：《中国抗生素杂志》2009年第1期18-23,60,共7页Chinese Journal of Antibiotics

基　　金：中国博士后项目(No.20070410356)。

摘　　要：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入到阿维链霉菌S-2(Streptomyces avermitilis 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 la。Objective To construct a Streptomyces avermitilis strain producing only “B” components by the replacement of aveD gene with a resistance cassette. Methods A DNA fragment carrying aveD gene amplified by PCR were interrupted by apr （apramycin resistance gene） inserted to a restriction site of NruI, the constructed recombinant 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 AveD24. 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 compounds were also identified as being oligomycin A and 5-oxoavermectin la by HPLC and LC-MS.

关 键 词：阿维链霉菌 AVERMECTIN aveD基因 寡霉素 5-酮基avermectin 

分 类 号：Q93[生物学—微生物学]