鸡Igλ轻链绿色荧光蛋白融合分子在COS7细胞膜的表达  

Expression of Chicken Ig Lambda Chain Fused with GFP on COS7 Cell Surface

作　　者：张红[1] 宋海涛[1] 张改平[2] 邢广旭[2] 赵东[2] 杨继飞[2] 柴书军[2] 郝青梅[3] 

机构地区：[1]河南农业大学,河南郑州450002 [2]河南省动物免疫学重点实验室,河南郑州450002 [3]河南教育学院,河南郑州450003

出　　处：《河南农业科学》2006年第11期102-104,109,共4页Journal of Henan Agricultural Sciences

基　　金：国家自然科学基金重点项目资助(30230270)

摘　　要：将牛IgG Fc受体γRⅡ的跨膜区序列嵌合入鸡Igλ轻链基因中,并将鸡Igλ轻链信号肽与Pegfp-C1载体的绿色荧光蛋白N端融合产生带有信号肽的中间载体Pegfp-C1-SP。鸡Igλ轻链基因嵌合分子定向克隆入载体的多克隆位点,构建了带有信号肽的鸡Igλ轻链/GFP融合蛋白真核表达载体。转染COS7细胞后,荧光显微镜观察,重组鸡Igλ轻链/GFP蛋白在COS7细胞膜及膜周围有明显表达,而载体pEGFP-C1转染的COS7细胞GFP则分布在细胞及细胞核。结果表明,牛IgG Fc受体γRⅡ的跨膜区能介导融合蛋白在细胞膜上的表达。Recombinant DNA encoding chimeric protein incorporating transmembrane region of bovine IgG Fc receptor γRⅡ （bFcRγRⅡ） and chicken Igγ light chain was generated, pEGFP- C1-SP, the vector with signal sequence was then created by fusing signal peptide of chicken Igγ to GFP N terminus. Cloned amplican was extracted using double restriction sites for cloning into modified pEGFP-C1-SP plasmid. DNA sequencing demonstrated that cloned amplican was inserted in frame and provided the desired coding sequence. Mammalian expression vector pEGFP -C1-SP-γR2T was successfully constructed. Expression of GFP fusion protein was located principally on the plasma membrane in COS7 cells transfected with pEGFP-C1-SP-）,R2T Vector as observed under fluorescent microscope. The results indicated that transmembrane region of bovine IgG Fc receptor γR Ⅱ was able to mediate plasma membrane targeting.

关 键 词：鸡Igγ轻链 牛IgG Fc受体γR Ⅱ跨膜区 COS7细胞 GFP融合蛋白 

分 类 号：S831[农业科学—畜牧学]