【摘要】 采用RTPCR技术从人肝总RNA中分离扩增了045kb的人铜锌超氧化物歧化酶(Cu/ZnSOD)基因的cDNA序列,首先克隆至大肠杆菌表达质粒pET23b,进行了序列测定和超高表达。将Cu/ZnSODcDNA亚克隆至乳酸乳球菌表达载体pMG36e,用电穿孔法将重组质粒pMG36esod转化到乳酸乳球菌中,获得Cu/ZnSOD的组成型表达,其表达量约占乳酸乳球菌可溶性蛋白的5%以上,活性染色表明该工程菌表达的Cu/ZnSOD具有较好的酶活性。更多还原

【Abstract】 The cDNA encoding human Cu/Zn SOD was amplified by RT PCR using the total RNA of human liver as the template,and was cloned into an E.coli expression vector pET23b.After the DNA sequence was determined,the recombinant plasmid pET23bsod was introduced into E.coli BL21(DE3)/pLysS.SDS PAGE analysis revealed that the recombinant E.coli expressed the predicted 19 kDa human Cu/Zn SOD,and its amount was over 50% of total proteins.The Cu/Zn SOD cDNA was then subcloned into a lactococcal expression vector pMG36e,and resulting pMG36esod was introduced into L.lactis MG1363 by electroporation.The human Cu/Zn SOD was expressed up to 5% of the soluble proteins,and the enzymatic activity was also observed by SOD activity dying.更多还原