武夷菌素产生菌CK-15遗传操作系统的优化

Optimization of Genetic Manipulation System for Wuyiencin Producer CK-15

旨在优化武夷菌素产生菌不吸水链霉菌武夷变种CK-15的遗传操作系统,实现武夷菌素生物合成基因的高效敲除。以自杀性质粒pKC1132为载体,优化了外源DNA通过接合转移进入菌株CK-15的方法,确立了构建基因敲除突变株的最佳方案,最终成功得到了武夷菌素生物合成基因的双交换突变株。获得了菌株CK-15稳定高效的遗传操作系统,为进一步研究武夷菌素生物合成机理和对武夷菌素利用组合生物合成改造奠定基础。

In order to optimize the genetic manipulation system of Streptomyces ahygroscopicus var. wuyiensis CK-15 for disrupting genes involved in wuyiencin biosynthesis, we developed a method of conjugation to transfer exotic DNA pKCl132 into S. ahygroscopicus var. wuyiensis CK-15. The best protocol of getting mutant by gene knockout was established and a double-crossover mutant ofwuyiencin biosynthesis gene was successfully obtained. A stable and efficient genetic manipulation system for S. ahygroscopicus var. wuyiensis CK-15 was developed, which was a solid foundation for the further study of wuyiencin biosynthesis mechanism and more important for combinatorial biosynthesis renovation of wuyiencin.