一种新型食品级表达载体pLEB590表达小鼠mCu/ZnSOD的初步研究  

Preliminary Study on Expression of mCu/ZnSOD by pLEB590,A Novel Food-grade Expression Vector

作　　者：罗立新[1] 王成[1] 施周铭[1] 

机构地区：[1]华南理工大学生物科学与工程学院,广东广州510006

出　　处：《食品与发酵工业》2009年第2期22-26,共5页Food and Fermentation Industries

基　　金：广东省农业攻关计划项目(2006B20501001)

摘　　要：采用RT-PCR技术从小鼠肝脏总RNA扩增0.46kb的小鼠铜锌超氧化物歧化酶基因的cDNA序列,首先T-A克隆至大肠杆菌表达质粒pUC19,进行序列测定。再将mCu/ZnSOD cDNA亚克隆至以nisⅠ为食品级选择标记的乳酸乳球菌表达载体pLEB590中,用电穿孔法将重组质粒pLEB590-mCu/ZnSOD转化到乳酸乳球菌MG1614中,经SDS-PAGE和Westernblotting检测,获得了mCu/ZnSOD的组成型表达,并通过SOD酶活测定表明该重组菌表达的mCu/ZnSOD具有较好的生物活性。The 0.46kb cDNA encoding Mus musculus Cu/Zn SOD（mCu/ZnSOD） was amplified by RT-PCR using the total RNA as the template from Mus musculus liver, and was cloned into an E. coli expression vector pUC19 to determine the DNA sequence. Then, the mCu/ZnSOD cDNA was subcloned into a lactococcat expression vector pLEBS90, choosing nisI as a food-grade selection marker, and the recombinant plasmid pLEB590-mCu/ZnSOD was transformed into L. lactis MG1614 by electroporation. The mCu/Zn- SOD was constitutive expressed in MG1614 as it was demonstrated by SDS-PAGE and Western blotting. And its good enzymatic activity was also observed by SOD activity assay.

关 键 词：乳酸乳球菌 pLEB590 食品级选择标记nisI 超氧化物歧化酶 

分 类 号：Q78[生物学—分子生物学]