钙结合蛋白D-9K基因原核表达载体的构建、表达产物纯化及其多克隆抗体的制备  

THE CONSTRUCTION OF PROKARYOTIC EXPRESSION VECTOR FOR THE GENE ENCODING THE CALCIUM BINDING PROTEIN D-9K, PURIFICATION OF THE EXPRESSED PRODUCT AND PREPARATION OF ITS POLYCLONAL ANTIBODY

作　　者：柯碧霞[1] 王健[2] 沈卫英[2] 高建民[1] 申庆祥[2] 

机构地区：[1]福建师范大学生物工程学院,福州350007 [2]上海市计划生育科学研究所,上海200032

出　　处：《细胞生物学杂志》2000年第3期135-140,共6页Chinese Journal of Cell Biology

基　　金：WHO资助项目(编号97826);国家重点基础研究规划项目(编号G1999055903)

摘　　要：本研究构建了pBV220/CabpD-9k-βhCG表达质粒,在大肠杆菌TG-1中进行表达,得到了CabpD-9k-βhCG融合蛋白。SDS-PAGE分析显示融合蛋白的分子量约为34.5KD。West-ern blot结果表明,融合蛋白与兔抗人hCGIgG特异结合。用纯化的融合蛋白免疫C57BL小鼠,制备了多克隆抗体,这为进一步研究钙结 合蛋白D-9k在着床中的功能和子宫内膜Ca^(2+)信号转导奠定了基础。The expression plasmid pBV220/CabpD-9K-βhCG was constructed,then was transferred into E.Coli TG-1 to express the fusion protein CabpD-9K-phCG. The result of SDS-PAGE showed that CabpD-9K-phCG had an apparent molecular weight of 34. 5KD. Western blot analysis indicated that the product CabpD-9K-|3hCG could be recognized specifically by the rabbit-anti-human hCG IgG. Finally we immunized C57 BL mouse with the purified fused protein to acquire the polyclonal antibody which would lay a foundation of the study on the CabpD-9K function and endometrial calcium signal transduction.

关 键 词：钙结合蛋白D-9K基因 原核表达 载体 

分 类 号：R392[医药卫生—免疫学]