重组人肝再生因子可溶性表达及活性鉴定  

Optimization of production process and activity identification of recombinant human liver regeneration factors

作　　者：李春男 毛羚羽 黄瑞麟 高慧英 岳鹏升 丁可盺 贾东明 崔春萍 LI Chun-nan;MAO Ling-yu;HUANG Rui-lin;GAO Hui-ying;YUE Peng-sheng;DING Ke-xin;JIA Dong-ming;CUI Chun-ping(Anhui Medical University,Hefei 230032,China;State Key Laboratory of Proteomics,Beijing Proteome Research Center,National Center for Protein Sciences Beijing,Beijing Institute of Lifeomics,Beijing 102206,China)

机构地区：[1]安徽医科大学,合肥230032 [2]北京蛋白质组研究中心蛋白质组学国家重点实验室国家蛋白质科学中心(北京)北京生命组学研究所,北京102206

出　　处：《军事医学》2018年第5期339-343,共5页Military Medical Sciences

基　　金：国家"重大新药创制"重大专项资助项目(2012ZX09102301-012)

摘　　要：目的探索重组人肝细胞生成素Cn(HPPCn)蛋白可溶性表达方法和纯化工艺,提高其比活性和蛋白产量,为发展新的急性肝病治疗药物奠定基础。方法利用分子生物学方法构建4个HPPCn原核表达载体,分别与不同的分子伴侣融合表达,助其表达后正确折叠。利用亲和层析方法纯化获得的融合蛋白,MTS法检测所得重组蛋白促进细胞增殖活性。结果成功构建了4个HPPCn表达载体,其中p MBP-P-HPPCn实现了rh HPPCn的可溶性表达。结论首次建立了重组HPPCn蛋白的可溶性生产方法,并对其蛋白活性进行分析和鉴定。Objective To explore the soluble expression method and purification process of recombinant human hepatopoietin Cn（HPPCn） protein in order to improve its specific activity and protein production, and contribute to the development of new therapeutic drugs for acute liver diseases. Methods We constructed four HPPCn prokaryotic expression vectors using molecular biology methods, and expressed them separately with different molecular chaperones to help them correctly fold after expression. The obtained fusion protein was purified by affinity chromatography, and the activity of the recombinant protein to promote cell proliferation was detected by MTS method. Results Four HPPCn expression vectors were successfully constructed, and the pMBP-P-HPPCn achieved soluble expression of rhHPPCn. Conclusion In this study, a soluble production method of recombinant HPPCn protein is established for the first time, and its protein activity is analyzed and identified.

关 键 词：肝细胞生长因子 肝细胞生成素Cn 重组蛋白质类 可溶性表达 MBP融合蛋白 蛋白纯化 肝细胞增殖 

分 类 号：R657.3[医药卫生—外科学]