HIV-1辅受体配体的融合表达载体的构建及表达  

Construction and Expression of the Bicistronic Expression Vector pCMV-R-K-S-K

作　　者：张颖[1] 白雪帆[1] 黄长形[1] 孙永涛[1] 潘蕾[1] 李光玉[1] 王临旭[1] 

机构地区：[1]第四军医大学唐都医院全军感染病诊疗中心,陕西西安710038

出　　处：《中国病毒学》2003年第1期23-26,T001,共5页Virologica Sinica

基　　金：国家自然科学基金项目(39970695)

摘　　要：应用PCR扩增RANTES-KDEL基因,鉴定后与真核表达载体pCMV—S/K连接,构建成HIV-1辅受体的配体、趋化因子RANTES和SDF-1的融合表达载体pCMV-R—K-S—K,酶切鉴定和测序证明成功构建了pCMV—R—K—S—K融合表达载体。脂质体介导pCMV—R—K-S—K转染HeLa细胞,间接免疫荧光证实RANTES和SDF-1可高效表达于HeLa细胞。结果表明构建的pCMV—R—K—S—K融合表达载体能在HeLa细胞中高效表达,可用于下一步的HIV-1感染实验。In order to construct bicistronic expression vector of RANTES and SDF-l,the ligands of HIV-1 principal coreceptors,RANTES-KDEL were amplified from pCMV-R-K by PCR and ligased with eukaryotic expression vector pCMV-S/K. The construction of pCMV-R-K-S-K was confirmed by enzymatic digestion and sequencing. Then pCMV-R-K-S-K transfection into HeLa cells was carried out by lipofectin. RAN TES and SDF-1 were showed expressed in HeLa cells by indirect immumofluo-rescence. The results show that pCMV-R-K-S-K was constructed and expressed in cell line HeLa successfully, which will contribute to gene therapy of HIV-1.

关 键 词：HIV-1 辅受体 趋化因子 融合表达载体 基因转染 

分 类 号：R346[医药卫生—基础医学]