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

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.