Optimizing expression of recombinant human monocyte chemoattractant protein-1 in E.coli
- VernacularTitle:重组人单核细胞趋化蛋白-1在大肠杆菌中表达的优化
- Author:
Hong MIAO
;
Baoyu GUO
;
Xu YANG
;
Pengqun YUAN
- Publication Type:Journal Article
- Keywords:
monocyte chemoattractant protein 1(MCP-1);
recombinant fusion proteins;
Escherichia coli;
orthogonal experimental design;
gene expression
- From:
Academic Journal of Second Military Medical University
2001;0(09):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To optimize the expression of recombinant human monocyte chemoattractant protein (rhuMCP-1) in E.coli DE3. Methods:With NBS-MICROS 15 L T.DR fermentor, pGEX-IN/huMCP-1 was constructed by our laboratory. Four parameters including pH,temperature,agitation rate and concentration of IPTG were studied by orthogonal experimental design. Results: It was found that the expression level was greatly affected by the amount of dissolved oxygen. This indicated that the agitation rate and ventilation amount were the most important parameters during fermentation. Examined by SDS-PAGE and gel scanning, the expression level of total protein was over 40% when agitation rate was 300 r/min and ventilation amount was 10 L/min. Conclusion: A method for high-level expression of huMCP-1 on pilot-scale is established, and it will be useful for large-scale industrial production of target protein.
