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本研究通过全化学法按大肠杆菌密码偏性合成了HBV PreS_2抗原决定簇基因,与ctxB基因的3’端融合。重组质粒转化大肠杆菌后融合基因得到高效表达,表达量达30μg/ml,表达产物95％以上分泌到胞外。表达的融合蛋白能与神经节苷脂GM1结合,说明融合蛋白保持了CTB的基本高级结构和生物学功能;ELISA实验证明融合蛋白具有CTB和HBV PreS_2的抗原性;应用亲和层析纯化后得到了电泳纯融合蛋白制品,为研究融合蛋白的免疫原性并进一步构建基因工程肽苗奠定了基础。
In this study, the HBV PreS_2 antigenic determinant gene was synthesized by the whole chemical method with the codon bias of E. coli and fused with the 3 ’end of ctxB gene. After the recombinant plasmid was transformed into E. coli, the fusion gene was highly expressed, with an expression level of 30 μg / ml, and over 95% of the expression product was secreted into the extracellular domain. The expressed fusion protein could bind to ganglioside GM1, indicating that the fusion protein retains the basic high-order structure and biological function of CTB. The results of ELISA showed that the fusion protein possessed the antigenicity of CTB and HBV PreS_2 and purified by affinity chromatography The electrophoresis pure fusion protein product, in order to study the immunogenicity of the fusion protein and further construction of genetic engineering peptide vaccine laid the foundation.