HIV-1 p24 gag 与hIL-2基因在重组痘苗病毒中的共表达研究  

Study on coexpression of HIV Ip24 gag and hIL 2 gene in recombinant vaccinia virus

作　　者：王玉红[1] 金宁一[1] 李体远 王洪军 王弘郡 刘素寰 李萍[2] 安汝国 殷震[2] 

机构地区：[1]白求恩医大基础医学院细胞生物教研室,长春130021 [2]解放军农牧大学研究所病毒室

出　　处：《白求恩医科大学学报》1999年第4期366-368,共3页Journal of Norman Bethune University of Medical Science

基　　金：中国杰出青年基金;国家自然科学基金

摘　　要：目的：研究 Ｈ Ｉ Ｖ１ｐ２４ ｇａｇ 与 ｈ Ｉ Ｌ２基因在重组痘苗病毒中的共表达。方法：以痘苗病毒复制非必需区血凝素（ Ｈ Ａ）基因为侧翼，将编码人白细胞介素２（ｈ Ｉ Ｌ２）的基因片段克隆到真核表达质粒ｐ１６ Ｑ Ｆ２４ Ｉ Ｌ２，经同源重组和血凝素阴性空斑筛选，获得了重组痘苗病毒ｖ１６ Ｑ Ｆ２４ Ｉ Ｌ２。结果：经间接免疫荧光试验、 Ｄｏｔ Ｅ Ｌ Ｉ Ｓ Ａ 和 Ｗ ｅｓｔｅｒｎ ｂｏｌｔ 等检测证明，重组病毒能同时表达ｐ２４ ｇａｇ 蛋白和 Ｉ Ｌ２蛋白，表达产物分子量分别为２４ ０００、１６ ０００。结论：这种重组痘苗病毒人表达外源蛋白的成功，为获得更强的免疫效果，研制 Ｈ Ｉ Ｖ 重组病毒活疫苗提供一种安全的新途径。Objective:Coexpression of HIV Ip24 gag and hIL 2 gene in recombinant vaccinia virus was studied.Methods:Coexpression plasmid p16QF24 IL2 containing cDNA of HIV Ip24 gag gene was constructed by inserting the hIL 2 gene under the p16QF1 24 plasmid vector with flank sequence of HA gene of vaccinia virus.After homologous recombination in Cos 7 cell,recombinant vaccinia viruses were purified in BHK21 cell by using plaque assay.Results:The recombinant vaccinia virus had coexpressed the HIV Ip24 gag and IL 2 protein in infected BHK21 and the molecular weight were about 24 kDa and 16 kDa by indirect immunofluorescence assay (IFA),Dot EIISA and Western blot.Conclusion:Coexpression is to produce better immune response in order to develop HIV vaccine.

关 键 词：HIV-Ip24gag HIL-2 痘苗病毒 

分 类 号：R392.2[医药卫生—免疫学]