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目的：将已构建好的新型抗乳腺癌双重靶向性融合蛋白Del1-9-G129R-T3的原核表达载体转化入原核表达宿主，诱导融合蛋白的表达，对之进行鉴定后进行分离纯化。方法：将融合蛋白重组表达质粒pET22b-Del1-9-G129R-T3转化入大肠杆菌BL21（DE3），IPTG诱导融合蛋白表达，收集菌体进行超声破碎，高速离心分离沉淀和上清。通过SDS-PAGE和Western blot对目的蛋白的表达进行检测和鉴定，利用亲和层析纯化融合蛋白。结果：SDS-PAGE结果显示，在诱导表达菌超声破碎后的沉淀物中出现一与预期分子量约24.7 kD大小相吻合的新蛋白区带，表明融合蛋白可能主要存在于细胞质中，形成不溶性包涵体。Western blot分析显示，蛋白在预期分子量处出现印迹。结论：融合蛋白Del1-9-G129R-T3已经在大肠杆菌中高效表达，为下一步融合蛋白的功能研究奠定基础。“,”Objective To transform the prokaryotic expression plasmid expressing a fusion protein of dual-targeting anti-breast cancer, Del1-9-G129R-T3, into E.coli expression host, and then induce and identify the expression of target protein. Methods The recombinant prokaryotic expression plasmid pET22b-Del1-9-G129R-T3 was first transformed into E.coli BL21(DE3). The expression of target protein was induced by IPTG. Bacteria were then col ected and ultrasonical y pulverized, fol owed by centrifugation and isolation of the precipitation and supernatant. Final y, analyzed and detected the expression of fusion protein by meas of SDS-PAGE and Western blot.The fusion protein was purified by affinity chromatography. Results SDS-PAGE results showed an new protein band in the bacterial precipitation. The size of the new protein was identical to the expected molecular weight of Del1-9-G129R-T3 (about 24.7 kD). These findings suggested that fusion protein may mainly locate in the cytoplasm and exist in the form of insoluble inclusion bodies. Western blot analysis demonstrated that the inducible protein appeared blot at expected molecular weight. Conclusion The Del1-9-G129R-T3 fusion protein was expressed in E.coli, which pave the way for further investigation on its function.