BCG HSP_(65)-GP_2融合蛋白的原核表达及其鉴定

Expression of fusion protein of BCG HSP_(65)-GP_2

目的 :表达可能对多种表皮来源的肿瘤具有预防及治疗作用的卡介苗热休克蛋白 6 5 (BCGHSP65)与人Her 2 /neuCTL表位 (GP2 )构成的融合蛋白 (BCGHSP65 GP2 )。方法 :将已构建的经筛选确定为正向重组的原核表达质粒PET15b BCGHSP65 GP2 转化感受态宿主菌BL2 1(DE3 )PlysS ,加IPTG诱导后 ,提取全菌总蛋白进行SDS PAGE分析 ,并用Western blot验证有无特异性目的蛋白表达。结果 :SDS PAGE分析显示 ,带正向重组质粒的菌株经IPTG诱导后 ,表达了一相对分子量约为 6 2 .3kD的新蛋白 ,Western blot结果显示诱导组具有特异性阳性条带。结论 :BCGHSP65 GP2 融合蛋白得到正确表达 ,使简便获得大量BCGHSP65 GP2 融合蛋白成为可能 ,为肿瘤疫苗的研究奠定了基础。

Objective:To investigate the expression of fusion protein of BCG HSP 65 combined with human Her 2/neu CTL epitope that fuctionally preventing and therapying from variant kind of epidermis tumor.Methods:PET 15b BCG HSP 65 GP 2 recombinant plasmid was transformed into BL 21 (DE 3)PlysS host bacterial strain.After IPTG inducing,the total protein was extracted and confirmed by SDS PAGE and Western blot.Results:It has been showed that a novel 62.3 kD protein was expressed by SDS PAGE analysis.The inducing group showed a specific positive band by Western blot analysis.Conclusions:It has been suggested that was expressed correctly.It is possible to obtain large scale BCG HSP 65 GP 2 fusion protein through simple approaches.It lays foundation on studies of tumor vaccine.