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目的构建弓形虫p22抗原基因克隆。方法参照文献合成引物,利用PCR技术获取P22基因片段,PCR产物经EcoRI、BamHI双酶切后,与经同样两种内切酶切割的质粒载体连接构成重组质粒PBluscript-SK-p22转化大肠杆菌DH5a,筛选鉴定重组子。结果PCR扩增及酶切分析均证实重组子含有插入的p22基因片段。结论成功的构建了弓形虫p22抗原基因克隆,为进一步选择适当的表达系统体外表达该抗原打下了必要的基础。
Objective To construct the Toxoplasma p22 antigen gene clone. Methods The P22 gene fragment was obtained by PCR. The PCR product was digested with EcoRI and BamHI and ligated with the plasmid vector cut by the same two endonucleases to construct the recombinant plasmid PBluscript-SK-p22 and transformed into E. coli DH5a , Screening identified recombinant. Results PCR amplification and digestion analysis confirmed that the recombinant contains the inserted p22 gene fragment. Conclusion The p22 antigen gene clone of Toxoplasma gondii was successfully constructed, which laid the necessary foundation for further selection of the appropriate expression system to express the antigen in vitro.