摘要
目的 构建含HIV 1中国流行株B亚型核心蛋白Gag基因的真核表达质粒 ,并在体外进行表达和鉴定。方法 将Gag基因插入到核酸疫苗载体质粒pVAX1中 ,构建真核表达质粒 pVAXGAG。用脂质体法 ,将重组质粒转染入Hela细胞后进行表达产物的检测。结果间接免疫荧光检测显示 ,转染重组质粒的细胞表面有绿色荧光。Westernblot和DotELISA分析显示 ,重组质粒转染细胞的裂解物中存在表达的Gag蛋白。 结论构建的核酸疫苗可在体外进行表达 。
Aim To construct the eukaryotic expression vector for HIV 1 CN core protein Gag gene and express it in vitro . Methods The recombinant expression vector pVAXGAG was constructed by inserting Gag gene into DNA vaccine vector pVAX1. The recombinant vector was transfected into Hela cell via liposome and the expressed product was detected by indirect immnuofluo rescence (IF),Western blot and Dot ELISA. Results The indirect IF showed green fluorescence on the membrane of transfected cells. Western blot and Dot ELISA demonstrated that expressed Gag protein existed in the lysate of Hela cells transfected by recombinant plasmid. Conclusion The constructed DNA vaccine can be expressed in vitro , and the expressed protein can react specifically with anti HIV P24 mAb.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2002年第6期531-532,共2页
Chinese Journal of Cellular and Molecular Immunology
基金
国家杰出青年基金资助No .3 982 5 119
