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目的 克隆并表达含有弓形虫P30抗原及霍乱毒素A2 /B亚基基因的原核表达载体 ,为弓形虫疫苗的研究奠定基础。方法 应用PCR方法扩增出P30基因片段后 ,克隆入含有霍乱毒素A2 /B亚基基因的表达质粒pUAB0 2 4 ,在大肠杆菌JM10 9(DE3)中表达融合蛋白。通过SDS -PAGE电泳及Westernblotting进行检测鉴定。 结果 电泳证明质粒构建正确 ,SDS -PAGE显示经IPTG诱导可以产生特异性条带 ,Westernblotting进一步证实该条带为P30 -CTA2 /B融合蛋白。结论 表达载体pUAB0 2 4 -P30可有效表达特异性的融合弓形虫抗原蛋白P30 -CTA2 B。
Objective To clone and express the prokaryotic expression vector containing Toxoplasma gondii P30 antigen and cholera toxin A2 / B subunit gene and lay the foundation for the study of Toxoplasma gondii vaccine. Methods After PCR amplification of P30 gene fragment, the expression plasmid pUAB024 containing the cholera toxin A2 / B subunit gene was cloned and the fusion protein was expressed in E. coli JM109 (DE3). By SDS-PAGE electrophoresis and Westernblotting detection and identification. The results of electrophoresis showed that the plasmid was constructed correctly. SDS-PAGE showed that specific bands could be induced by IPTG. Western blotting confirmed that the band was P30-CTA2 / B fusion protein. Conclusion The expression vector pUAB0 2 4 -P30 can effectively express the specific fusion antigen Toxoplasma gondii P30-CTA2 B.