DNA表达载体构建及HIV多表位核酸疫苗的设计与表达  

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

作　　者：付延军[1] 金宁一[2] 金洪涛[2] 李臻[2] 辛苏文[2] 李萍[2] 韩贞珍[1] 李太元[1] 

机构地区：[1]延边大学农学院,龙井133400 [2]军事医学科学院军事兽医研究所病毒室,长春130062

出　　处：《中国生物制品学杂志》2007年第4期237-239,共3页Chinese Journal of Biologicals

基　　金：军队科技攻关重点项目(06G127);"863"项目(2006AA02Z447)资助

摘　　要：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。Objective To construct a novel eukaryotie expression vector based on Semliki forest virus（SFV） replieon and express the DNA vaccine containing subtype core protein p24 and muhi-epitope MEG chimeric gene of epidemic strain B of HIV-1 in China. Methods Digest plasmid pSFV-MCS with restriction endonuelease, and insert the obtained SFV replieon into vector plRESneo containing CMV promoter and enheneer to construct a novel eukaryotie expression vector pCS. Insert HIV-1 MEGp24 gene into vector pCS, then transfeet the constructed recombinant DNA vaccine pCS-MEGp24 to BHK-21 cells in mediation of liposome. Identify the expressed preduet by IFA. Results IFA proved that HIV-1 MEGp24 was expressed in transfeeted 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 preduet showed good specificity. It provided an important experimental basis for the feasibility of developing novel DNA vaccine against HIV-1 strain epidemic in China.

关 键 词：人免疫缺陷病毒 塞姆利基森林病毒 复制子 核酸疫苗 

分 类 号：Q786[生物学—分子生物学] R512.91[医药卫生—内科学]