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.

This article focused on basic clinical skill training of Changhai Hospital,in such aspects as courses,teaching infrastructure,faculty competence,teaching pattern,evaluation system and incentives mechanism.These efforts aim at exploring the establishment of a new clinical skills teaching model,which prove satisfactory and improving the standardized residents basic clinical skill training system.