| 重组人MASP2蛋白在大肠杆菌中的表达和纯化 |
| |
| 引用本文： | 高婷,赵怀龙,刘萱,曹诚. 重组人MASP2蛋白在大肠杆菌中的表达和纯化[J]. 生物技术通讯, 2011, 22(6): 806-808,891. DOI: 10.3969/j.issn.1009-0002.2011.06.013 |
| |
| 作者姓名： | 高婷 赵怀龙 刘萱 曹诚 |
| |
| 作者单位： | 军事医学科学院生物工程研究所,北京,100850 |
| |
| 摘 要： | 目的:获得酶原形式的重组人甘露聚糖结合凝集素相关丝氨酸蛋白酶2(MASP2)。方法:在大肠杆菌中诱导表达重组人MASP2全长蛋白,包涵体裂解后,经复性、透析、浓缩、考马斯亮蓝染色、SDS-PAGE及Western印迹,鉴定纯化结果及酶活性。结果:复性后的MASP2蛋白经考马斯亮蓝染色未见杂带。自激活实验表明,当MASP2浓度在1μmool/L以下时,无论在4℃还是37℃,都能较稳定地保持酶原形式;蛋白浓度为3.5μmool/L时只能在4℃保持稳定,37℃发生自激活;蛋白浓度达到12μmool/L后,在4℃时已不能稳定存在。结论:获得了较纯的重组人MASP2蛋白,且具有自激活活性。
|
| 关 键 词： | 甘露聚糖结合凝集素相关丝氨酸蛋白酶 原核表达 包涵体 复性 自激活 |
Expression in Escherichia coli and Purification of Full Length Recombinant Human MASP2 |
| |
| GA Ting,ZHA Huai-Long,LIU Xuan,CA Cheng. Expression in Escherichia coli and Purification of Full Length Recombinant Human MASP2[J]. Letters in Biotechnology, 2011, 22(6): 806-808,891. DOI: 10.3969/j.issn.1009-0002.2011.06.013 |
| |
| Authors: | GA Ting ZHA Huai-Long LIU Xuan CA Cheng |
| |
| Affiliation: | GAO Ting,ZHAO Huai-Long,LIU Xuan,CAO Cheng Beijing Institute of Biotechnology,Beijing 100850,China |
| |
| Abstract: | Objective: To express in Escherichia coli and purify the zymogenic recombinant human mannose-binding lectin associated serine protease 2(MASP2).Methods: The expression of full length wild type recombinant human MASP2 was induced by IPTG in Escherichia coli.The inclusion bodies were solubilized,and diluted into refolding buffers.The renatured proteins were dialyzed against PBS,pooled and concentrated,and estimated by SDS-PAGE,Coomassie blue stain and Western blot.Results: The refolded protein was detected by... |
| |
| Keywords: | mannose-binding lectin associated serine protease prokaryotic expression inclusion body autoactivating |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ |
|
