HIV-1 B亚型gag基因密码子优化及其免疫原性的研究

从河南HIV-1流行区感染者中克隆HIV-1 B亚型gag基因,通过序列比对获得其一致性共有序列,对该共有序列按照哺乳动物优势密码子的使用原则进行优化,以Western blot方法比较优化前后gag基因体外表达量.发现对gag基因进行密码子优化可显著提高其表达水平.将优化后的mod.gag基因插入重组腺病毒载体,构建了重组病毒rAdV-mod.gag.在BALB/c小鼠体内分别以108PFIJ及108PFU rAdV-mod.gag疫苗单独免疫两次均可产生较高水平的gag特异性细胞免疫反应.由此得出结论,对gag基因的密码子优化是成功的;表达优化后gag基因的重组腺病毒疫苗,可以在小鼠体内诱导较强的gag基因特异性CTL应答.

Codon Modification of HIV-1 Subtype B gag Gene and Its Immunogenicity in Mice

HIV-1 subtype B gag genes were cloned from the infected paid blood donors in Henan, and the consensus sequence based on these prevalent strains was obtained by aligning. The codons of the consensus gag sequence were modified according to mammalian codon usage. Western blot analysis was used to compare the expression level of wild type and codon-modified gag gene. It was found that the expression level of Gag protein was improved largely by codon-modification. Then the mod.gag gene was inserted into the adenovirus vector and the recombinant adenovirus rAdV-mod.gag was constructed. 108 PFU or 109 PFU rAdV-mod.gag vaccinated mice twicely could elicit high level Gag-specific CTL responses in immunized mice. In conclusion,the codon modification of gag gene is successful. The recombinant adenovirus vaccine harbouring mod.gag can induce robust Gag-specific CTL immune response in mice.