【摘要】 以福建红曲菌M-CL为出发菌株,以PSKH质粒为模板克隆PtrpC启动子及TrpC终止子。通过重叠延伸PCR技术融合PtrpC启动子、MpigE (目的)基因、TrpC终止子,获得PMT融合片段。为了验证融合片段PMT的正确性,进行琼脂糖凝胶电泳分析,在2 100bp左右出现单一且清晰的条带。通过EcoR V和SpeI酶切PMT片段,插入带有潮霉素抗性的PSKH质粒的EcoR V和Spe I位点,构建1个红曲菌过表达载体HPMT,大小为7 155bp。通过构建过表达载体,以期对红曲菌M-CL进行改造,获得高产色素低产桔霉素的菌株。更多还原

【Abstract】 By taking the M-CL of Monascusin Fujian as the original strain,the PtrpCpromoter and TrpCterminator were cloned by using the PSKH plasmid as the template.The PMT fusion fragments were obtained by fusing PtrpC promoter,MpigEtarget gene and TrpCterminator with the overlap-extension PCR technology.In order to verify the correctness of the fusion fragment PMT,the agarose gel electrophoresis analysis was carried out,and a single clear band appeared at about 2,100 bp.The fragment of PMT was digested by EcoR V and SpeI,and the EcoR V and Spe I sites of the PSKH plasmid with hydthromycin resistance were inserted to construct an overexpression vector HPMT of Monascus,with a size of 7 155 bp.By constructing the overexpression vector,the M-CL of Monascus was modified,thus to obtain the strain with high yield of pigment and low yield of citrinin.更多还原