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.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
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)~~
关键词
可溶性补体1型受体
表达
包含体
纯化
复性
soluble complement receptor type 1
expression
inclusion body
purification
refolding