一株表达延胡索酸水合酶的基因工程菌的构建  

The Construction of an E.coli Strain with Expression of Fumarate Hydratase

作　　者：史燕东[1] 吴梧桐[1] 张玉彬[1] 王旻[1] 龚韬 

机构地区：[1]南京中国药科大学生物技术中心

出　　处：《中国药科大学学报》1994年第3期159-162,共4页Journal of China Pharmaceutical University

摘　　要：应用ＰＣＲ技术从大肠杆菌染色体ＤＮＡ中扩增出了长约１．８ｋｂ的ｆｕｍＡ基因片段，将其插入到ｐＵＣ１９载体中，转化大肠杆菌ＪＭ１０５，最后筛选出一株延胡索酸水合酶高产菌，命名为ＣＰＵ－Ｗ－９２１１，该菌的延胡索酸水合酶活力比ＪＭ１０５提高约６倍。从ＣＰＵ－Ｗ－９２１１里提取的质粒ｐＵＦ５２作为ＰＣＲ扩增的模板，以合成的ｆｕｍＡ基因两侧各长２４个核苷酸的片段为引物，可以扩增出１．８ｋｂ的ｆｕｍＡ片段；用限制性内切酶也可以从ｐＵＦ５２中切出一个接近１．８ｋｂ的插入片段。An 1.8 kb fumA fragment,amplified out from E.coli chromosome DNA by PCR technology,was inserted into plasmid pUC19.The recombinant plasmid was used to transform E.coli JM105.From the transformants,a strain, named CPU-W-9211 was obtained,whose activity of fumarate hydratase is five times higher than that of the host strain JM105.Plasmid pUF52 extracted from CPU-W-9211 is about 1.8kb longer than plasmid pUC19.With pUF52 as template DNA,the 1.8 kb fumA fragment can also be obtained by PCR technology.

关 键 词：PRC 重组 延胡索酸水 酶 

分 类 号：TQ464.8[化学工程—制药化工]