HIV-1 DNA疫苗的构建及免疫原性

目的构建稳定表达HIV-1Gag-Pol蛋白的DNA疫苗,提高目的基因的表达效率。方法对HIV-1野生型Gag-Pol基因序列进行优化,使用本室构建的表达载体VR,构建重组质粒VR-GPCINS。将构建的重组质粒VR-GPCINS转染HEK293细胞,Western blot鉴定表达产物。用构建的DNA疫苗免疫BALB/c小鼠,Western blot检测体液免疫反应。结果成功构建能稳定高效表达HIV-1Gag-Pol蛋白的VR-GPCINS质粒,Western blot在免疫小鼠的血清中检测到抗p24的特异性抗体。结论含优化Gag-Pol基因序列的质粒DNA疫苗VR-GPCINS在细胞表达和体液免疫方面均明显优于含野生型Gag-Pol基因序列的质粒DNA疫苗VR-GP。

Construction and Immunogenicity of HIV-1 DNA Vaccine

Objective To construct a recombinant plasmid for stable expression of HIV-1 Gag-Pol protein,which may be used as a DNA vaccine.Methods The Gag-Pol gene sequence was amplified from wild HIV-1,then modified and inserted into constructed VR vector.Transfect HEK293 cells with the obtained recombinant plasmid VR-GPCINS and identify the expressed product by Western blot.Immunize BALB/c mice with the recombinant plasmid VR-GPCINS and determine the antibody in sera by Western blot.Results Western blot proved the expression of HIV Gag-Pol in HEK293 cells transfected with recombinant plasmid VR-GPCINS.The specific antibody against p24 was found in the sera of mice immunized with the recombinant plasmid.Conclusion Compared with the recombinant plasmid VR-GP containing Gag-Pol gene sequence of wild HIV-1,the recombinant plasmid VR-GPCINS containing modified Gag-Pol gene sequence was more suitable for expression in eukaryotic cells,and more effective in inducing humoral immune response.