Sanglifehrin A产生菌Streptomyces flaveolus DSM 9954遗传转化系统的建立  

The genetic system development of Sanglifehrin A producing host Streptomyces flaveolus DSM 9954

作　　者：刘运[1] 陶科[1] 史冠莹[1] 瞿旭东[2] 侯太平[1] 

机构地区：[1]四川大学生命科学学院生物资源与生态环境教育部重点实验室,成都610064 [2]中国科学院上海有机化学研究所生命有机化学国家重点实验室,上海200032

出　　处：《四川大学学报（自然科学版）》2011年第2期441-444,共4页Journal of Sichuan University(Natural Science Edition)

基　　金：国家自然科学基金(30800034)

摘　　要：Sanglifehrin A的产生菌淡黄色链霉菌Streptomyces flaveolus DSM 9954,通过筛选,得到其菌丝培养基为TSB,生孢培养基为ISP-4,抗性实验显示淡黄色链霉菌Streptomycesflaveolus DSM 9954对所选抗生素都较敏感.利用原生质体转化和接合转移都成功地构建了两种遗传转移系统,并且从接合转移成功的链霉菌转化子中回收质粒pKC 1139,经EcoRⅠ单酶切电泳,验证了质粒确实通过接合转移从大肠杆菌转移至宿主链霉菌,这为研究Sanglifehrin A的生物合成机制和结构改造奠定了基础.Sanglifehrin A is produced by Streptomyces flaveolus DSM 9954. TSB culture was selected as its growing culture, ISP-4 as sporing culture. Antibiotic resisting test showed S. flaveolus DSM 9954 was sensitive to all the chosen antibiotics. The genetic transformation system of S. flaveolus DSM 9954 was successfully developed either by the protoplast transformation or conjugation between S. flaveolus and E. coll. And the plasmid pKCl139 was rescued from the transformants and confirmed by EcoR I digestion, which proved the success of conjugation between S. flaveolus and E. coll. These results laid the foundation for studying Sanglifehrin A＇s biosynthesis pathway and producing its analogues.

关 键 词：Sanglifehrin A 原生质体转化 接合转移 

分 类 号：Q78[生物学—分子生物学]