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目的构建弓形虫微线体MIC8的真核表达重组质粒。方法设计合成弓形虫MIC8引物,运用PCR方法扩增其基因片段克隆至真核表达质粒pCDNA3.1,从而构建重组表达质粒pCDNA3.1-MIC8。结果 PCR扩增弓形虫MIC8基因序列正确,构建的重组表达质粒pCDNA3.1-MIC8经PCR、HindⅢ和XbaI双酶切和测序鉴定正确。结论成功获得真核表达重组质粒pCDNA3.1-MIC8,为进一步研究弓形虫疫苗的免疫保护性奠定基础。
Objective To construct the eukaryotic expression recombinant plasmid of Toxoplasma gondii MIC8. Methods Toxoplasma gondii MIC8 primers were designed and synthesized. The gene fragment was amplified by PCR and cloned into eukaryotic expression plasmid pCDNA3.1 to construct recombinant expression plasmid pCDNA3.1-MIC8. Results The Toxoplasma gondii MIC8 gene was amplified by PCR. The constructed recombinant plasmid pCDNA3.1-MIC8 was identified by PCR, Hind Ⅲ and XbaI digestion and sequencing. Conclusion The eukaryotic recombinant plasmid pCDNA3.1-MIC8 was successfully obtained, which laid the foundation for further study on the protective immunity of Toxoplasma gondii vaccine.