中国动物传染病学报Issue(2)：58-64,7.
鸭疫里默氏杆菌GroEL蛋白的原核表达和单克隆抗体制备
GENERATION OF MONOCLONAL ANTIBODIES TO GROEL OF RIEMERELLAANATIPESTIFER
摘要
Abstract
The objective of this study was to generate monoclonal antibodies (MAbs) against GroEL protein of Riemerellaanatipestifer. The GroEL gene was cloned from R. anatipestifer strain WJ4 and the recombinant GroEL was expressed in E. coli. After purification, the recombinant GroEL was used to immunize BALB/c mice 3 times. Spleen cells of immunized mice were fused with murine myeloma SP2/O cells. Specific MAb secreting hybridomas were screened using indirect ELISA and positive hybridomas were cloned 3 times. As a result, two hybridoma cell lines (1G2B6, 1G2F10) stably producing anti-GroEL MAbs were identified and established. The isotype of both MAbs was IgG1. The ELISA titer of ascetic fluids was determined to be 1:102 400. In Western blot, both MAbs reacted with R. anatipestifer serotypes 1, 2 and 10 strains but not with Escherichia coli, Salmonella enterica and Pasteurella multocida strains. Besides, both MAbs were specific for R. anatipestifer serotypes 1, 2 and 10 strains tested. The availability of MAbs specific for GroEL paved a path for further development of diagnostic kit for R.anatipestifer.关键词
鸭疫里默氏杆菌/GroEL蛋白/单克隆抗体Key words
Riemerellaanatipestifer/GroEL/monoclonal antibody分类
农业科技引用本文复制引用
侯湾湾,韩先干,王少辉,王小兰,岳嘉蘋,曹守林,范红结,于圣青..鸭疫里默氏杆菌GroEL蛋白的原核表达和单克隆抗体制备[J].中国动物传染病学报,2014,(2):58-64,7.基金项目
公益性行业（农业）科研专项经费项目（201003012）；国家自然科学基金 （）
