| 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: 原核表达 重组蛋白 包涵体 |
| DOI： |
| 分类号: |
| 基金项目:国家自然科学基金项目（30370383） |
|
| Prokaryotic expression and purification of recombinant protein soluble HLA-G1 |
|
|
| 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. |
| Key words: HLA-G prokaryotic expression recombinant protein inclusion body |
