重组人载脂蛋白AI_(米兰变体)在大肠杆菌中的高效表达  

Over-expression of Proapolipoprotein AI_(Milano) in Escherichia coli

作　　者：丁满生[1] 马文峰[1] 张梅芳 刘大涛 郭美锦[1] 庄英萍[1] 储炬[1] 龚邦强 

机构地区：[1]华东理工大学生物反应器国家重点实验室,上海200237 [2]上海凯曼生物科技有限公司,上海200233 [3]上海信谊药业有限公司药研所,上海200234

出　　处：《微生物学通报》2005年第4期107-111,共5页Microbiology China

基　　金：国家高技术研究发展计划项目("863"项目)(No.2002AA217021);国家重大科技专项(No.2002AA2Z3451)

摘　　要：通过大肠杆菌表达系统来生产重组人载脂蛋白AIM(proapolipoprote in AIM,proapoAIM)。整个proapo AIM基因被分成两个片断,通过RT-PCR的方法合成。在对该基因的上游部分进行改造后,插入到表达载体pBV220中进行表达。蛋白的表达量达到45%左右,蛋白的表达形式为包涵体。包涵体通过疏水柱进行柱上复性,复性后的蛋白具有良好的生物活性。To facilitate the mutation of Milano site, the whole proapo AIM gene was divided to two fragments to synthesize by RT-PCR. The optimization of upstream gene sequence was carried out to improve proapo AIM expression level in E. coll. After induction with a shift of temperature, yields of recombinant proapo AIM achieved about 45 % of total cell protein and the recombinant proapo AIM was expressed as a form of inclusion body in cells. The renaturation of protein was carried with a hydrophobic interaction column, the results showed that recomhinant proapo AIM had similar functional properties identical to those of native protein.

关 键 词：载脂蛋白AI MILANO 包涵体 疏水柱 复性 

分 类 号：Q819[生物学—生物工程]