重组人可溶性补体受体1型SCR15-18片段表达纯化及生物活性鉴定  

Expression,purification and bioactivity identification of recombinant SCR15-18 domain of human soluble complement receptor type 1

作　　者：谭兵[1] 张德纯[2] 汪正清[1] 

机构地区：[1]第三军医大学基础医学部微生物学教研室 [2]重庆医科大学基础医学院病原生物学教研室

出　　处：《第三军医大学学报》2007年第10期892-895,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金(30471723)~~

摘　　要：目的获得高表达、高纯度和高活性的可溶性补体受体1型SCR15-18(sCR1-SCR15-18)蛋白。方法重组原核表达载体pET32a-sCR1-SCR15-18在大肠杆菌BL21中经不同IPTG浓度、诱导时间和温度诱导表达,超声破菌,提取包含体,经Ni2+-NTA亲和层析后,选择不同氧化还原条件进行复性,进而检测其生物学活性。结果得到了有较高表达量、较高纯度和较好生物学活性的sCR1-SCR-15-18蛋白。结论优化了sCR1-SCR15-18蛋白的表达、纯化和复性的参数,所获结果为进一步动物体内保护实验研究奠定了基础。Objective To prepare highly expressed, purified and refolded SCR15-18 of human soluble complement receptor type 1 （ sCR1-SCR15-18 ） protein. Methods The expression of recombinant pET32- sCR1-SCR15-18 in E. coli.. BL21 was induced by IPTG of different concentrations for different time period under different temperatures and the bacteria were split by sonication. The sCR1-SCR15-18 protein was purified by Ni^2＋ -NTA resin affinity chromatography. The purified protein was refolded under different conditions. Then the bioactivity of the protein was analyzed. Results The sCR1-SCR15-18 protein of high expression, purity and bioactivity was attained. Conclusion The parameters of expression, purification and refolding of sCR1- SCR15-18 protein were optimized, which may pave a way for further studies.

关 键 词：可溶性补体1型受体 表达 包含体 纯化 复性 

分 类 号：R392.33[医药卫生—免疫学] R392.11[医药卫生—基础医学]