【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。更多还原

【Abstract】 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.更多还原