Position: Home > Articles > Expression,purification and activity indentification of HLA-G1
Journal of Northwest A & F University(Natural Science Edition)
2004,32
(7)
16-20
HLA-G1的基因表达、纯化及其活性分析
作 者：
汤海洋;张彦明;汪蕴;刘亚兵;丰美福
单 位：
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;西北农林科技大学动物科技学院
关键词：
HLA-G1;原核表达;蛋白纯化;融合蛋白;蛋白活性
摘 要：
在获得HLA-G1cDNA克隆序列的基础上,构建了pGEX-4T2-HLA-G1原核表达载体,对HLA-G1的诱导表达发现,融合蛋白以包涵体形式存在于表达菌中。进一步溶解包涵体,改善复性条件,最终获得高纯度的GST/HLA-G1融和蛋白。51Gr释放试验表明,纯化的HLA-G1具有抑制NK细胞对靶细胞的杀伤活性。
译 名：
Expression,purification and activity indentification of HLA-G1
作 者：
TANG Hai-yang~(1,2),ZHANG Yan-ming~1,WANG Yun~2,LIU Ya-bing~2,FENG Mei-fu~2(1 College of Animal Science and Technology,Northwest Sci-Tech University of Agriculture and Forestry,Yangling,Shaanxi 712100,China;f Zoology,Chinese Academy of Sciences,Beijing 100080,China)
关键词：
HLA-G;prokaryotic expression;protein purification;fusion protein;protein activity
摘 要：
HLA-G,which is a non-classical HLA class I molecule,plays an important role of immunotolerance.Based on the cloning of HLA-G1 cDNA,we constructed prokaryotic expression plasmid,pGEX-4T2-sHLA-G1,and transformed into E.coli BL21(DE3).GST fusion proteins highly expressed in E.coli after being induced by IPTG,but they formed inclusion bodies which have no native structures and no biological activities.So we had to lyse and renature them for the further research.Finally,we got purified GST/HLA-G fusion protein which can inhibit the cytotoxicity of NK cell.
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