曲张链菌素产生菌壮观链霉菌NRRL2494菌株遗传操作体系的建立  

Gene Manipulation System for Streptovaricin Producer Streptomyces spectabilis NRRL2494

作　　者：宋姣姣[1] 康前进[2] 张连茹[1] 吴莹莹[1,3] 白林泉[2] 

机构地区：[1]厦门大学生命科学学院,天然产物源靶向药物国家地方联合工程实验室,福建厦门361102 [2]上海交通大学生命科学技术学院,微生物代谢国家重点实验室,上海200240 [3]中国科学院上海生命科学研究院植物生理生态研究所,合成生物学重点实验室,上海200032

出　　处：《厦门大学学报（自然科学版）》2016年第1期60-66,共7页Journal of Xiamen University：Natural Science

基　　金：国家自然科学基金(31100027);高等学校博士学科点专项科研基金(20110121120013)

摘　　要：曲张链菌素属于安莎类抗生素,具有显著的生物活性.在从壮观链霉菌NRRL2494菌株的发酵产物中分离和鉴定了多组分曲张链菌素的基础上,探索和优化了外源DNA通过接合转移进入壮观链霉菌NRRL2494菌株的操作方法和培养条件.以游离型质粒pJTU1278为载体,在体外构建了一个曲张链菌素酰胺合酶基因svaF敲除的质粒,通过接合转移转入到壮观链霉菌NRRL2494野生型菌株中,所获得的svaF基因缺失突变株失去了产生曲张链菌素的能力.该遗传操作体系的成功建立和优化,使得在体内分析和鉴定曲张链菌素生物合成基因的功能成为可能,同时也为建立其他类似放线菌的遗传操作体系提供了参考.In order to enable the streptovaricin biosynthetic study by in vivo gene disruption, it is crucial to develop a genetic modification system for the streptovaricin producer Streptornyces spectabilis NRRL2494. In this study,we aimed to establish a method of conjugation to transfer exogenous DNA into Streptomyces spectabilis NRRL2494. A putative streptovaricin amide synthase gene svaF was disrupted in vitro ,and the constructed plasmid was transferred into Streptomyees spectabilis NRRL2494 by conjugation through a double-crossover recombination event.The resulting svaF mutant lost the ability to produce streptovaricin. Based on the above work,we developed a genetic manipulation system for Streptomyces spectabilis NRRL2494, enabling the functional characterization of streptovaricin biosynthetic genes in vivo ,and offering a positive example for other fast-growing Actinobacteria which lack an appropriate genetic manipulation system.

关 键 词：壮观链霉菌 遗传操作体系 曲张链菌素 生物合成 酰胺合酶 

分 类 号：Q933[生物学—微生物学]