【摘要】 目的 :探讨HIV 2gag非融合蛋白的表达。方法 :应用DNA重组技术 ,将HIVgag基因全序列 (gag)和部分 (gag’)cDNA片段克隆到pBV2 2 0载体PRPL 串联启动子下游 ,构建成HIV 2gag重组表达载体pBV gag和pBV gag’ ,在大肠杆菌中表达。结果 :SDS PAGE显示pBV gag’在 2 1kD处可见一明显的额外蛋白带 ,经薄层扫描分析占菌体总蛋白的 6 .1% ,蛋白印迹结果证明 ,表达出的特异蛋白分子量pBV gag为 5 7kD和 47kD ;pBV gag’为 47kD和 2 1kD。经提取包涵体证明 ,表达的蛋白主要以包涵体形式存在于菌体中。结论 :HIV 2gag非融合蛋白可在大肠杆菌中获得表达并主要以包涵体形式存在更多还原

【Abstract】 Objective:In order to study the expression of HIV2 gag gene in E.coli.Methods:Construction of recombinant expression plasmid by digesting the plasmid vector pCR TMII containing HIV2 gag gene with BgⅡ.And the fragments of gag containing full longth of HIV2 gag gene and gag’containing HIV2 gag gene truncated at Cterminus were obtained and inserted downstram of P RP L promoter on a plasmid pBV220,and then expressed in E.coli DH5α.Results: The molecular weight of expressed proteins were 57 kD and 47 kD (gag),47 kD and 21 kD(gag’) respectively.Only gag’showed additional band at 21 kD on SDSPAGE.Densitometric analysis showed that the content of 6.1%in total E.Coli proteins.Western blot result showed that however,all calculated molecular weight of gag and gag’proteins were specific reactions with positive sera of HIV2 patients.The expression products of gag and gag’ gene were presented in inclusion bodies.Conclusion:HIV2 gag genes expressed in E.Coli DH5α and mainly presented in inclusion bodies.更多还原