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目的克隆空肠弯曲菌penner o:19 cjaA基因,构建其原核表达载体,诱导表达及纯化蛋白,为基因工程疫苗的研制打下基础。方法扩增空肠弯曲菌penner o:19 cjaA基因,构建pGEM-T-cjaA克隆载体,以EcoRI和XhoI为酶切位点构建pGEX-4T-1-cjaA原核表达载体。表达载体转化E.coli BL21后诱导表达,SDS-PAGE鉴定表达产物,用Glutathione Sepharose4B纯化重组蛋白。结果成功构建pGEX-4T-1-cjaA表达载体,并获得浓度为10μg/μl的rGST-CjaA重组蛋白。结论成功构建pGEM-T-cjaA克隆载体及pGEX-4T-1-cjaA表达载体,获得重组蛋白rGST-CjaA,为研究空肠弯曲菌疫苗打下基础。
Objective To clone Penicillium o: 19 cjaA gene of Campylobacter jejuni, construct its prokaryotic expression vector, induce expression and purify the protein, laying a foundation for the development of genetically engineered vaccine. Methods The Penicillium o: 19 cjaA gene of Campylobacter jejuni was amplified and cloned into pGEM-T-cjaA vector. The prokaryotic expression vector pGEX-4T-1-cjaA was constructed by EcoRI and XhoI restriction enzyme digestion. The expression vector was transformed into E. coli BL21 and induced to express. The expressed product was identified by SDS-PAGE and the recombinant protein was purified with Glutathione Sepharose 4B. Results The pGEX-4T-1-cjaA expression vector was successfully constructed and rGST-CjaA recombinant protein was obtained at a concentration of 10μg / μl. Conclusion The pGEM-T-cjaA cloning vector and pGEX-4T-1-cjaA expression vector were successfully constructed, and the recombinant protein rGST-CjaA was obtained, which laid the foundation for the study of Campylobacter jejuni vaccine.