|
|||||
|
|
| 重组蛋白sHLA-G1的原核表达与纯化 | |
| 引用本文： | 张 劲,郭霭光,丰美满.重组蛋白sHLA-G1的原核表达与纯化[J].西北农林科技大学学报(社会科学版),2007,35(8):180-184. |
| 作者姓名： | 张 劲 郭霭光 丰美满 |
| 作者单位： | 1. 西北农林科技大学,生命科学学院,陕西,杨凌,712100;中国科学院,动物研究所,生物膜与膜生物工程国家重点实验室,北京,100080 2. 西北农林科技大学,生命科学学院,陕西,杨凌,712100 3. 中国科学院,动物研究所,生物膜与膜生物工程国家重点实验室,北京,100080 |
| 摘 要： | 为了进一步在蛋白质水平对人白细胞抗原HLA-G进行分析,利用已经构建好的克隆质粒pGEM-T-HLA-G1,进一步构建了重组原核表达质粒pET28a-sHLA-G1。SDS-PAGE检测表明,重组蛋白sHLA-G1在大肠杆菌(E.coli)中得到稳定表达,原核表达的重组蛋白sHLA-G1主要以包涵体形式存在。包涵体蛋白经充分洗涤、尿素溶解后用钴离子树脂纯化,并用SDS-PAGE和Western印迹进行了分析,鉴定结果表明,该蛋白纯度达到90%以上,并能与HLA-G特异性抗体87G反应。该研究结果为进一步的复性工作奠定了基础。 |
| 关 键 词： | HLA-G 原核表达 重组蛋白 包涵体 |
| 文章编号： | 1671-9387（2007）08-0180-05 |
| 收稿时间： | 2006/7/14 0:00:00 |
| 修稿时间： | 2006-07-14 |
Prokaryotic expression and purification of recombinant protein soluble HLA-G1 |
|
| ZHANG Jin,GUO Ai-guang,FENG Mei-fu.Prokaryotic expression and purification of recombinant protein soluble HLA-G1[J].Journal of Northwest Sci-Tech Univ of Agr and,2007,35(8):180-184. | |
| Authors: | ZHANG Jin GUO Ai-guang FENG Mei-fu |
| Institution: | 1 College of Life Sciences,Northwest A & F University,YangLing,Shaanxi 712100,China;2 The Laboratory of Biomembrane and Membrane Biotechnology,Institute of Zoology,The Chinese Academy of Sciences,Beijing 100080,China) |
| Abstract: | In order to study human leukocyte antigen(HLA-G) protein,on the basis of a recombinant cloning plasmid pGEM-T-HLA-G1,prokaryotic expression plasmid pET28a-sHLA-G1 was constructed,which could express soluble recombinant protein HLA-G1 in Escherichia coli(E.coli).Most of His-sHLA-G1 existed in the inclusion body.The inclusion body was washed extensively and solubilized with urea,and then purified with Co2 resin.Finally,the purified protein was analyzed by using SDS-PAGE and Western blot.The results indicated that the purity of His-sHLA-G1 could reach 90%,and the recombinant protein could react with monoclonal antigen 87G.This research provided the basis for the further refolding study on HLA-G molecule. |
| Keywords: | HLA-G |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ | |
| 点击此处可从《西北农林科技大学学报(社会科学版)》浏览原始摘要信息 | |
| 点击此处可从《西北农林科技大学学报(社会科学版)》下载免费的PDF全文 | |