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目的:分析弓形虫致密颗粒蛋白GRA8真核表达质粒的体外表达。方法:大量提取纯化重组真核表达质粒pVAX1-GRA8,瞬时转染培养的vero E-6细胞,RT-PCR和免疫印迹法检测GRA8在细胞中的表达。结果:从转染pVAX1-GRA8后的vero E-6细胞的RNA中扩增出GRA8的特异条带,转染细胞存在弓形虫感染血清识别的特异蛋白质条带。结论:真核表达质粒pVAX1-GRA8在vero E6细胞中获得表达。
Objective: To analyze the in vitro expression of Toxoplasma gondii GRA8 eukaryotic expression plasmid. Methods: The recombinant eukaryotic expression plasmid pVAX1-GRA8 was extracted and purified. The cultured vero E-6 cells were transiently transfected. The expression of GRA8 was detected by RT-PCR and Western blot. Results: The specific bands of GRA8 were amplified from the RNA of vero E-6 cells transfected with pVAX1-GRA8, and the specific protein bands recognized by Toxoplasma gondii infection sera were present in the transfected cells. Conclusion: The eukaryotic expression plasmid pVAX1-GRA8 is expressed in vero E6 cells.