pPRMETb-TRAIL原核表达载体的构建和表达及纯化  

The Construction, Expression and Purification of Prokaryotic Expression Vector pPRMETb-TRAIL

作　　者：孙等军[1] 方琴[1] 陈剑[2] 肖芸[2] 任志娟[2] 张维[2] 张旭[2] 王季石[2] 

机构地区：[1]贵阳医学院附院药剂科,贵州贵阳550004 [2]贵阳医学院附院血液内科,贵州贵阳550004

出　　处：《贵阳医学院学报》2002年第1期4-6,10,共4页Journal of Guiyang Medical College

基　　金：贵州省省长专项资金资助项目(S2 0 0 1 - 1 4 )

摘　　要：目的 :为获取较高纯度的肿瘤坏死因子相关凋亡诱导配体 (TNF relatedapoptosis inducingligand ,TRAIL)工程蛋白。方法 :将TRAIL基因构建于原核表达载体pPRMETb中 ,在大肠杆菌中由异丙基硫代 β D 半乳糖苷 (IPTG)诱导表达 ,对包涵体进行洗涤、尿素裂解、复性后过镍柱层析纯化。结果 :经酶切、测序鉴定证实 pPRMETb TRAIL构建完全正确 ,在大肠杆菌xl1 blue中经IPTG诱导表达量较高 ,包涵体经尿素裂解后行镍柱纯化得到了较高纯度的TRAIL蛋白。结论Objective: To abtain a pure TRAIL protein. Methods: A prokaryotic expression vector pPRMETb TRAIL was constructed and induced by IPTG to express in E.coli. TRAIL protein was purified with nickel column chromatography after inclusion body lysis , protein renaturation and recovery . Results:The constrution of prokaryotic expression vector pPRMETb TRAIL was proofed correct by digestive identification and sequence measuring. TRAIL protein induced by IPTG had a high expression in xl1 blue. A pure TRAIL protein was obtained with nickel column chromatography after inclusion body lysis. Conclusion: A technological procedure of obtaining a pure TRAIL protein was successfully established.

关 键 词：大肠杆菌 细胞包涵体 聚合酶链反应 肿瘤坏死因子相关凋亡诱导配体 pPRMETb-TRAIL 原核表达载体 

分 类 号：Q782[生物学—分子生物学] R378.21[医药卫生—病原生物学]