| 大肠杆菌中可溶性表达重组人成纤维细胞生长因子21及制备 |
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| 引用本文： | 范开,谭克海,张淳,张益.大肠杆菌中可溶性表达重组人成纤维细胞生长因子21及制备[J].重庆工学院学报,2011(5). |
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| 作者姓名： | 范开 谭克海 张淳 张益 |
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| 作者单位： | 重庆理工大学药学与生物工程学院; |
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| 基金项目： | 国家科技支撑计划资助项目(2008DAI63D05) |
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| 摘 要： | 为在大肠杆菌中高效、非融合、可溶性表达重组人成纤维细胞生长因子21(rhFGF-21),研究了纯化方法,以获得具有较高生物活性的rhFGF-21。全基因合成人成纤维细胞生长因子21cDNA序列,克隆进原核表达载体pET-3c,在转化大肠杆菌BL21(DE3)PlysS中进行表达。表达产物经盐析、层析等方法纯化后,利用3T3-L1细胞鉴定其促葡萄糖摄取的生物学活性。结果表明:rhFGF-21在BL21(DE3)PlysS实现了高效、非融合、可溶性表达,目的蛋白占菌体总蛋白的20%左右;表达产物经纯化后,纯度可达95%以上;表达产物能够明显促进3T3-L1细胞对葡萄糖的吸收。该方法成功实现了rhFGF-21在大肠杆菌中高效可溶性表达。
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| 关 键 词： | 人成纤维细胞生长因子21 可溶性表达 葡萄糖摄取 |
Soluble Expression and Preparation of Recombinant Human Fibroblast Growth Factor 21 in E.coli |
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| FAN Kai,TAN Ke-hai,ZHANG Chun,ZHANG Yi.Soluble Expression and Preparation of Recombinant Human Fibroblast Growth Factor 21 in E.coli[J].Journal of Chongqing Institute of Technology,2011(5). |
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| Authors: | FAN Kai TAN Ke-hai ZHANG Chun ZHANG Yi |
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| Affiliation: | FAN Kai,TAN Ke-hai,ZHANG Chun,ZHANG Yi(School of Pharmacy and Bioengineering,Chongqing University of Technology,Chongqing 400054,China) |
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| Abstract: | To effectively express human Fibroblast Growth Factor 21 in the form of solubility without fusion and purify it from E.coli,the gene of human FGF-21 was integrated and inserted into pET-3c,and itconstructed the recombinant vector pET-3c-FGF21 which was subsequently transformed into BL21(DE3)PlysS.The engineering bacteria was induced by IPTG to express recombinant human FGF-21.The recombinant protein's biological function of uptake glucose on 3T3-L1 was tested after purifying with centrifugation,salting and ... |
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| Keywords: | human FGF-21 soluble expression glucose uptake |
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