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目的:构建人n IL-2与n HBsAg融合基因并转化到BCG中构建重组BCG(recombinant BCG，rBCG)，为制备治疗和预防乙型肝炎和结核病的二联疫苗奠定基础。n 方法:以IL-2 pMalLP质粒和HBsAg pMalLP质粒为模板，设计引物，利用PCR技术扩增n HBsAg和n IL-2基因，进行重叠PCR获得融合基因n IL-2-n HBsAg并纯化，将融合基因克隆至穿梭表达载体pMV261得到重组质粒，电转化导入BCG感受态细胞，以蛋白质印迹法检测融合基因在rBCG中的表达。n 结果:PCR扩增得到n IL-2和n HBsAg，通过重叠PCR获得n IL-2-n HBsAg融合基因后成功插入穿梭表达载体，rBCG能表达融合蛋白。n 结论:构建人n IL-2与n HBsAg融合基因重组质粒并导入BCG，获得了稳定表达融合蛋白的rBCG。n “,”Objective:To construct the fusion gene of human n IL-2 and n HBsAg and transform into BCG for expression, laying foundation for preparation of bivalent vaccine for prevention and treatment of hepatitis B and tuberculosis.n Methods:Using IL-2 pMalLP and HBsAg pMalLP as templates, primers were designed. n HBsAg and n IL-2 genes were amplified by PCR, and then the fusion gene n IL-2-n HBsAg was obtained by overlapping PCR and purified. The fusion gene was cloned on the shuttle expression vector pMV261 to obtain the recombinant plasmid, and then transformed into BCG sensitive cells by electroporation, and the expression of the fusion gene in recombinant BCG (rBCG) was detected by Western blot.n Results:IL-2 (453 bp) gene and n HBsAg (618 bp) gene were amplified by PCR, and n IL-2-n HBsAg (1 179 bp) fusion gene was amplified by overlapping PCR. The fusion gene was successfully inserted into the shuttle expression vector. The fusion protein was detected in rBCG.n Conclusion:The recombinant plasmid of human n IL-2 and n HBsAg fusion gene is successfully constructed and introduced into BCG to obtain rBCG which can stably express the fusion protein.n