BALB/c小鼠I-A^dαβ链RFP融合双顺反子表达载体的构建及其在COS-7中的表达  

Construction and expression of pRed-IRES-I-A^d in COS-7 cell line

作　　者：高明[1] 王海平[1] 王燕宁[2] 周勇[1] 王全立[1] 

机构地区：[1]军事医学科学院野战输血研究所,北京100850 [2]兰州军区兰州总医院输血科,兰州730050

出　　处：《生物技术通讯》2004年第6期546-548,共3页Letters in Biotechnology

基　　金：国家自然科学基金资助项目(30200245)

摘　　要：构建了在β链C端融合红色荧光蛋白(RFP)标签的BALB/c小鼠I-Adαβ链真核双顺反子表达载体pRed-IRES-I-Ad,使用LipofectAMINE2000转染COS-7细胞,用激光共聚焦显微镜观察外源蛋白在细胞中的表达与定位。I-Adαβ分子在COS-7细胞中能够以较高的效率表达,并且在COS细胞中能形成聚集状态。与通常的真核翻译帽子结构起始相比,IRES启动真核翻译系统的效率低于前者;与空质粒对比,IRES介导的真核翻译起始,RFP的表达量较低。A bicistronic eukaryotic vector expressing BALB/c mice I-Ad αβ chains simultaneously was constructed. RFP encoding gene was fused to the C terminus of β chain. COS-7 cell line was transfected with the plasmid by using LipofectAMINE2000. The expression of target gene was detected by laser confocus microscope. The foreign gene can be expressed efficiently in COS-7 cell line. Laser confocus microscope results showed the protein located in endogenous membrane system. Compare with cap-dependent translation, cap-independent IRES is less efficient in translation initiation.

关 键 词：IRES RFP 真核 BALB/C小鼠 表达载体 COS-7细胞 激光共聚焦显微镜 红色荧光蛋白 外源蛋白 表达量 

分 类 号：Q782[生物学—分子生物学] R512.63[医药卫生—内科学]