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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 express a fusion protein (BCGHSP65 GP2) consisting of the BCGHSP65 and human Her2 / neuCTL epitopes (GP2) that may have preventive and therapeutic effects on a variety of epidermal-derived tumors. METHODS: The constructed recombinant prokaryotic expression plasmid PET15b BCGHSP65 GP2 was transformed into competent cells BL21 (DE3) PlysS. After induction with IPTG, total proteins were extracted and analyzed by SDS PAGE. Western Blot Verify that there is no specific target protein expression. Results: SDS PAGE analysis showed that a new recombinant plasmid with a relative molecular weight of about 6.23 kD was induced by IPTG. Western blot results showed that the induced group had a specific positive band. Conclusion: The correct expression of BCGHSP65 GP2 fusion protein makes it easy to obtain a large amount of BCGHSP65 GP2 fusion protein, which lays the foundation for the study of tumor vaccine.