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目的:空肠弯曲菌鞭毛抗原FlaA蛋白人工表达及免疫原性的分析。方法:使用PCR方法获得flaA编码的基因,经确证后将该基因克隆到谷胱甘肽转移酶融合表达载体(pGEX-5x-1)中,构建了pGEX-flaA表达质粒,在大肠杆菌BL21中获得了相应表达,并采用Glutathione Sepharose4B进行纯化后获得目的蛋白,用该蛋白免疫小鼠制备了特异性抗血清,利用经培养获得CJ菌体的裂解液做为抗原,通过用最基本的免疫凝集及免疫扩散方法对该抗血清进行初步的分析。结果:pGEX-flaA重组菌株具有明显的表达带,其分子量与预期的结果相同,该表达蛋白占总蛋白的20%,使用裂解的CJ抗原粗纯液,免疫凝集试验观察阳性。结论:利用基因克隆-原核表达所获得FlaA融合蛋白是具有相应的免疫原性的。
Objective: To analyze the artificial expression of FlaA protein of Campylobacter jejuni antigen and its immunogenicity. Methods: The flaA gene was obtained by PCR. After confirmed, the gene was cloned into glutathione transferase fusion expression vector (pGEX-5x-1), and the pGEX-flaA expression plasmid was constructed and expressed in E. coli BL21 Obtained the corresponding expression, and purified by Glutathione Sepharose 4B to obtain the target protein, the mouse immunized with the protein to prepare a specific antiserum, CJ cell lysate obtained by culturing as an antigen, by using the most basic immune agglutination And immunodiffusion method of the antiserum for a preliminary analysis. Results: The recombinant strain pGEX-flaA had a significant expression band with the same molecular weight as the expected result. The expressed protein accounted for 20% of the total protein. The crude CJ antigen was lysed, and the immunohistochemistry was positive. Conclusion: FlaA fusion protein obtained by gene cloning and prokaryotic expression is correspondingly immunogenic.