摘要
在大肠杆菌中，利用pET－3c表达载体，高效表达了具有全长序列的HIV—1整合酶蛋白（p31）。表达产物主要以包涵体的形式存在，用1mol／LNaCl溶解包涵体可部分地溶解p31蛋白，溶解上清中p31蛋白具有很高的纯度。本文进一步描述了一种在变性条件下重组蛋白的纯化方法，即将包涵体用8mol／L尿素溶解，先后经过S-SepharoseFastFlow与Sephacryl-300柱层析，可将重组产物纯化到电泳纯。纯化蛋白以1μg／ml包被ELISA板，分析正常人及HIV－1阳性个体的血清，结果重组p31与检测的所有41份阳性血清均可发生特异反应，而与36份正常血清则不起反应，表明纯化的重组蛋白可用于检测血清中的抗HIV-1抗体。
The Sequence encwhng the fall length Of human inununodchciency virus type I integrase ha been cloned and highly expand becdy using PET system in E. colt. The recombinant product mainly existed in inclusion bodies (IB). Quantities Of ~inant integraSe Protein could be released by dissolving the inclusion babies with I cool/L NaCI, resulting in high delve Of pndcallon. alternatively, IB Protein could be solublized by & mol/L urea and puritied under denaturing conditions. The prDduct has been poded close to homogeneity sequenhally by S-Sepharose Fast Flow and Sephacryl-300 columns. The poded Product reacted with all 41 HIV-1 posihve sera but none Of 36 normal sera tested. This suggests that the recolnbinant integrase protein aught be useful for detechng HIV-1 anhbodies in blood samples.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
1999年第1期17-20,共4页
Chinese Journal of Cellular and Molecular Immunology
基金
"九五"国家医学科技攻关资助!96-906-03-16
