【摘要】 目的:构建HIV-1重组腺病毒疫苗并初步鉴定其免疫原性。方法:用Adeno-X Expression System试剂盒将HIV-1 gagpol基因片段插入到腺病毒载体上,通过脂质体介导将获得的重组腺病毒质粒Adeno-X-gagpol转染至293细胞,使之自主包装成有感染活性的重组腺病毒rAd-gagpol,用western-blotting法鉴定其表达情况;用CsCl密度梯度离心法纯化该重组腺病毒,以5×108pfu/mL的滴度1 mL单次免疫小鼠,15天后测小鼠体液免疫效果。结果:克隆序列与设计相符,重组腺病毒疫苗能稳定表达gagpol蛋白,体液免疫中检测出抗p55和p24的特异性抗体。结论:HIV-1 gagpol重组腺病毒构建成功,具有一定的体液免疫原性。更多还原

【Abstract】 Objective: To construct a recombinant adenovirus vaccine which carried HIV-1 gagpol gene and to study its immunogenicity to prevent HIV-1 virus. Methods: By Adeno-X Expression System Kit, an Adeno-X-gagpol plasmid was constructed, by inserting HIV-gagpol gene into recombinant adenvirus genome. The contructed plasmid was transfected into 293 cell by lipofectamine 2000,and western-blotting were used to detection protein expression. The recombinant adenvirus was purified by CsCl and 1 mL of the plasmid with concentration of 5×108pfu/mL was injected into BALB/c mouse, then western-blotting was used to detection its immunogenicity after 15 days. Results: The recombinant adenovirus was constructed successfully, and could express HIV-1 gagpol protein efficiently.Specific anti-p55 antibody and anti-p24 antibody were detected in sera from mice immunized with recombinant adenovirus vaccine. Conclusion: The HIV-gagpol recombinant adenovirus vaccine with good humoral immunogenicity has been established successfully.更多还原