猪传染性胸膜肺炎放线杆菌APX Ⅱ基因特异片段的克隆和表达  

Cloning and expressing of Actinobacillus pleuropneumonia APX Ⅱ gene

作　　者：袁秀芳[1] 王一成[1] 徐丽华[1] 张存[1] 叶伟成[1] 

机构地区：[1]浙江省农业科学院畜牧兽医研究所,浙江杭州310021

出　　处：《浙江农业学报》2007年第1期6-10,共5页Acta Agriculturae Zhejiangensis

基　　金：浙江省科技计划重点项目(2004C22044)

摘　　要：将猪传染性胸膜肺炎放线杆菌APXⅡ的一段毒素基因克隆到原核表达载体pET-28a(+)的多克隆位点中,经鉴定后得到重组质粒pET-APXⅡ,并将其转化到受体菌BL21中,诱导剂IPTG进行诱导表达,4 h后达到高峰。经12%SDS-PAGE电泳检测,表达得到的融合蛋白大小约为31 kD。经Western blotting分析,表达蛋白能与APP阳性血清发生特异性反应,而与阴性血清不反应,说明该表达蛋白具有免疫原性,这为下一步用此表达蛋白作为抗原,对APP进行诊断试剂的研制和基因工程疫苗的研究提供科学依据。The fragment ofActinobacillus pleuropneumonia APXⅡ gene was cloned into the multiple cloning site of Eukytote expressing vector pET-28a, The recombinant of pET- APXⅡ was cultivated and induced with IPTG. The amount of expressed protein reached peak at 4 hours after the induction. The results of SDS-PAGE and Western blotting showed that the protein was 31 kD in size and specifically reacted with APP-positive sera from pigs and rabbits, not negative sera, indicating that the recombinant protein had high specificity of immune reaction and can be used as antigen of diagnostic assay and gene processing vaccine for APP in practice.

关 键 词：猪传染性胸膜肺炎放线杆菌 克隆 表达 Westemblotting 

分 类 号：S858.28[农业科学—临床兽医学]