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| 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 | |
| 引用本文： | 谭兵,张德纯,汪正清.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[J].第三军医大学学报,2007,29(10):892-895. |
| 作者姓名： | 谭兵 张德纯 汪正清 |
| 作者单位： | 1. 第三军医大学基础医学部微生物学教研室,重庆市微生物工程实验室,重庆,400038;重庆医科大学基础医学院病原生物学教研室,临床检验诊断学省部共建教育部重点实验室,重庆,400016 2. 重庆医科大学基础医学院病原生物学教研室,临床检验诊断学省部共建教育部重点实验室,重庆,400016 3. 第三军医大学基础医学部微生物学教研室,重庆市微生物工程实验室,重庆,400038 |
| 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. |
| 关 键 词： | 可溶性补体1型受体 表达 包含体 纯化 复性 |
| 文章编号： | 1000-5404（2007）10-0892-04 |
| 修稿时间： | 2006-09-25 |
Expression, purification and bioactivity identification of recombinant SCR15-18 domain of human soluble complement receptor type 1 |
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| TAN Bing,ZHANG De-chun,WANG Zheng-qing.Expression, purification and bioactivity identification of recombinant SCR15-18 domain of human soluble complement receptor type 1[J].Acta Academiae Medicinae Militaris Tertiae,2007,29(10):892-895. | |
| Authors: | TAN Bing ZHANG De-chun WANG Zheng-qing |
| Institution: | 1. Department of Microbiology, College of Medicine, Third Military Medical University, Chongqing 400038, 2.Key Laboratory of Laboratory Medical Diagnostics of Ministry of Education, Chongqing Medical University, Chongqing 400016, China |
| Abstract: | 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 Ni2 -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. |
| Keywords: | soluble complement receptor type 1 expression inclusion body purification refolding |
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