鸡Igλ轻链基因克隆、序列分析及其在COS7细胞膜上的定位表达  

Cloning and Expression of the Chicken Igλ Light Chain on COS7 Plasma Membrane

作　　者：张红[1,2,3] 王健[3] 王丽[2] 席俊[2] 易明林[2] 张改平[2] 章金刚[1] 

机构地区：[1]军事医学科学院,北京100850 [2]河南省农业科学院农业部动物免疫学重点开放实验室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》2009年第8期132-136,共5页Journal of Henan Agricultural Sciences

基　　金：国家"863"计划项目(2006AA10Z446);国家自然科学基金项目(30571388);河南省高校创新人才支持计划(2009HASTIT035);河南省基础与前沿项目(072300430110)

摘　　要：为了研究鸡B细胞膜免疫球蛋白(mIg)的功能,深入了解鸡免疫系统抗体基因的特点,从鸡法氏囊B细胞cDNA中扩增出Igλ轻链基因,对其核苷酸序列和氨基酸序列进行了分析和比较。该基因cDNA全长873bp,编码含226个氨基酸的Igλ轻链,N-端21个氨基酸构成轻链信号肽,随后是2个Ig样结构域。运用融合PCR的方法将该基因与牛IgG Fc受体γRⅡ跨膜区序列嵌合形成重组跨膜分子,构建真核表达载体pcDNA-λR2T,转染COS7细胞,荧光抗体染色及流式细胞术检测到重组鸡Igλ轻链在细胞膜上的表达。所扩增的鸡Igλ轻链基因以及构建表达于细胞膜上的重组鸡Igλ轻链跨膜分子,为研究鸡免疫系统中的抗体轻链基因,探索Igλ轻链和B细胞膜免疫球蛋白的功能奠定技术基础。To study the functions of chicken antibody genes, a chicken Ig）, light chain gene was amplified from chicken bursa B lymphocyte eDNA pool. The gene spans 873bp of DNA which en- codes a chicken Igλ light chain with 226 amino acids. The light chain consists of a signal peptide with 21aa at N-terminus and two antibody-like domains. Using fuse PCR technique, the recombi- nant transmembrane molecule was formed by generating a chimeric protein incorporating trans- membrane region of bovine IgG Fc receptor λR Ⅱ （bFcRλR Ⅱ ） and chicken Ig）, light chain. Mam- malian expression vector pcDNA-）,R2T was successfully constructed and transiently transfected into COS7 cells via lipofectamin. Flow cytometric analysis showed that the expression of the chi- meric Igλ light chain was located principally on the plasma membrane on COS7 cells. The gene cloning and membrane-located expression of the chicken Igλ, light chain provide a basis for further study of chicken antibodies and B cell membrane immunoglobulin.

关 键 词：鸡Igλ轻链 重组跨膜分子 COS7细胞 膜定位表达 牛IgGFc受体γRⅡ 

分 类 号：Q78[生物学—分子生物学]