DNA表达载体构建及HIV多表位核酸疫苗的设计与表达

目的利用塞姆利基森林病毒(SFV)复制子构建新型真核表达载体,并对含HIV-1中国流行株B亚型核心蛋白p24及多表位MEG嵌合基因的核酸疫苗进行表达与鉴定。方法将含SFV复制子的元件从pSFV-MCS中切下,连入含CMV启动子及增强子的pIRESneo载体中,构建新型真核表达载体pCS,再将含HIV-1 MEGp24基因插入至pCS载体中,构建重组核酸疫苗pCS-MEGp24。用脂质体法将重组质粒转染入BHK-21细胞,进行表达产物的检测。结果间接免疫荧光检测显示,重组质粒转染的BHK-21细胞能有效表达HIV-1 MEGp24,并与HIV阳性血清反应。结论所构建的核酸疫苗可在BHK-21细胞系内进行表达,且MEGp24基因的表达蛋白具有特异性,为构建新型HIV-1中国流行株核酸疫苗的可行性提供了重要实验依据。

Construction of DNA Expression Vector and Design and Expression of HIV-1 Multi-epitope DNA Vaccine

Objective To construct a novel eukaryotic expression vector based on Semliki forest virus(SFV) replicon and express the DNA vaccine containing subtype core protein p24 and multi-epitope MEG chimeric gene of epidemic strain B of HIV-1 in China.Methods Digest plasmid pSFV-MCS with restriction endonuclease,and insert the obtained SFV replicon into vector pIRESneo containing CMV promoter and enhancer to construct a novel eukaryotic expression vector pCS.Insert HIV-1 MEGp24 gene into vector pCS,then transfect the constructed recombinant DNA vaccine pCS-MEGp24 to BHK-21 cells in mediation of liposome.Identify the expressed product by IFA.Results IFA proved that HIV-1 MEGp24 was expressed in transfected BHK-21 cells and reacted with HIV-positive serum.Conclusion The constructed DNA vaccine pCS-MEGp24 was expressed in BHK-21 cells,and the expressed product showed good specificity.It provided an important experimental basis for the feasibility of developing novel DNA vaccine against HIV-1 strain epidemic in China.