刺糖多孢菌鼠李糖和福乐糖胺合成基因的克隆和组装  

Cloning and reconstitution of rhamose and forosamine biosynthetic gene of Saccharopolyspora spinosa NRRL18395

作　　者：郭航[1] 白亭丽[1] 陶美凤[1,2] 

机构地区：[1]华中农业大学农业微生物学国家重点实验室,武汉430070 [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200030

出　　处：《华中农业大学学报》2012年第3期298-302,共5页Journal of Huazhong Agricultural University

基　　金：国家自然科学基金项目(30870073)

摘　　要：鼠李糖和福乐糖胺是多杀菌素生物合成必需的2个脱氧糖结构单元,负责其合成的4种酶的基因(gtt、epi、gdh、kre)与多杀菌素生物合成基因簇并不在一起,而是分散分布在染色体的3个位点上。为克隆到完整的多杀菌素生物合成基因簇,采用构建基因组文库、PCR扩增等手段从刺糖多孢菌NRRL18395染色体分别克隆gtt、epi、gdh和kre基因,并将其克隆组装在同一整合型载体上,以便于构建用于表达多杀菌素糖单元合成基因的表达载体。Rhamnose and forosamine are two essential deoxysugars components of spinosad.Four genes involved in the biosynthesis of rhamnose and forosamine,i.e.gtt（encoding NDP-glucose synthase）,gdh（encoding NDP-glucose dehydratase）,epi（encoding epimerase） and kre（encoding 4′-ketoreductase）,are not located together with the spinosad biosynthetic gene cluster,but distributed in 3 scattered loci in the chromosome of Saccharopolyspora spinosa NRRL18395.To get an integral spinosad biosynthetic pathway,the four genes were cloned by constructing a genomic library and PCR amplifying from S.spinosa NRRL18395.The 4 genes were successively assembled into an integrative vector to construct an expression vector for biosynthesis of rhamnose and forosamine in heterologous hosts.

关 键 词：刺糖多孢菌 多杀菌素 鼠李糖 福乐糖胺 cosmid文库 原位杂交 

分 类 号：Q78[生物学—分子生物学]