阿维链霉菌BAC文库的构建及分析

Construction and analysis of a BAC library of Streptomyces avermitilis genome

为进一步阐明阿维菌素的生物合成路径和调控途径,提高阿维菌素的产量,采用BamHⅠ限制性内切酶酶解阿维链霉菌基因组DNA的方法,构建了阿维链霉菌的细菌人工染色体(BAC)文库。该文库共包含2 304个克隆,平均插入片段为101kb,空载率约9%,覆盖了该菌基因组的25.9倍。该文库的成功构建可以为其他微生物尤其是基因组具有磷硫酰化修饰的微生物的文库构建提供参考。

To further clarify the biosynthesis and regulation pathways of avermectins and to increase its production,a bacterial artificial chromosome(BAC)library was constructed using the BamHⅠ-digested genomic DNA of Streptomycesavermitilis.The Streptomyces avermitilis BAC library was consisted of 2 304 clones with an average insert size of 101 kb,empty clone rate of about 9%,and 25.9-fold coverage of the whole genome.The DNA phosphorothioation of the Streptomyces avermitilis genome made it very difficult to construct a BAC library.The successful construction of this BAC library will provide a reference for constructing BAC libraries of other microorganisms,especially those with DNA phosphorothioation.