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目的:构建含有分子佐剂CD40L的真核表达质粒hCGβ-TT,以提高hCGβ的免疫原性。方法:用分段PCR的方法构建了hCGβ-TT,然后插入双元真核表达质粒IRES的A位点,将CD40L插入pIRES的B位点构建hCGβ-TT-IRES-CD40L。结果:经测序所克隆的基因hCGβ-TT和CD40L序列正确。结论:利用分子佐剂CD40L与hCGβ-TT的连接,构建了hCGβ-TT-IRES-CD40L真核细胞表达质粒,为检测hCGβDNA免疫避孕疫苗的免疫原性及抗生育作用奠定了基础。
Objective: To construct eukaryotic expression plasmid hCGβ-TT containing molecular adjuvant CD40L to enhance the immunogenicity of hCGβ. Methods: hCGβ-TT was constructed by segmented PCR and inserted into the A site of the binary eukaryotic expression plasmid IRES. CD40L was inserted into the B site of pIRES to construct hCGβ-TT-IRES-CD40L. Results: The sequence of cloned genes hCGβ-TT and CD40L was correct. CONCLUSION: The eukaryotic expression plasmid hCGβ-TT-IRES-CD40L was constructed by linking CD40L with hCGβ-TT, which laid the foundation for detecting the immunogenicity and anti-fertility of hCGβ DNA immunocontraceptive vaccine.