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| 重组人载脂蛋白AI米兰变体在大肠杆菌中的高效表达 | |
| 引用本文： | 丁满生,马文峰,张梅芳,刘大涛,郭美锦,庄英萍,储炬,龚邦强.重组人载脂蛋白AI米兰变体在大肠杆菌中的高效表达[J].微生物学通报,2005,32(4):107-111. |
| 作者姓名： | 丁满生 马文峰 张梅芳 刘大涛 郭美锦 庄英萍 储炬 龚邦强 |
| 作者单位： | 1. 华东理工大学生物反应器国家重点实验室,上海,200237;上海凯曼生物科技有限公司,上海,200233 2. 华东理工大学生物反应器国家重点实验室,上海,200237 3. 上海凯曼生物科技有限公司,上海,200233 4. 上海信谊药业有限公司药研所,上海,200234 |
| 基金项目： | 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。 |
| 关 键 词： | 载脂蛋白AI Milano 包涵体 疏水柱 复性 |
| 文章编号： | 0253-2654(2005)04-0107-05 |
| 收稿时间： | 10 19 2004 12:00AM |
| 修稿时间： | 11 22 2004 12:00AM |
Over-expression of Proapolipoprotein AIMilano in Escherichia coli |
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| Ding ManSheng;Ma WenFeng;Zhang MeiFang;Liu DaTao;Guo MeiJin;Zhuang YingPing;Chu Ju;Gong BangJiang.Over-expression of Proapolipoprotein AIMilano in Escherichia coli[J].Microbiology,2005,32(4):107-111. | |
| Authors: | Ding ManSheng;Ma WenFeng;Zhang MeiFang;Liu DaTao;Guo MeiJin;Zhuang YingPing;Chu Ju;Gong BangJiang |
| Abstract: | To facilitate the mutation of Milano site, the whole proapo AI_(M) gene was divided to two fragments to synthesize by RT-PCR. The optimization of upstream gene sequence was carried out to improve proapo AI_(M) expression level in E. coli. After induction with a shift of temperature, yields of recombinant proapo AI_(M) achieved about 45 % of total cell protein and the recombinant proapo AI_(M) 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 recombinant proapo AI_(M) had similar functional properties identical to those of native protein. |
| Keywords: | Proapolipoprotein AI Milano Inclusion body Hydrophobic Renaturation |
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