猪源D型产毒素多杀性巴氏杆菌毒素基因的原核表达  

Prokaryotic Expression of ToxA Gene of Swine Toxigenic Pasteurella Multocida Serotype D

作　　者：江涛[1,2] 谭春萍[1] 任灵芝 黄百花[1] 刘文娟[1] 陆芹章[1] 

机构地区：[1]广西大学动物科学技术学院,广西南宁530005 [2]固始县动物疫病预防控制中心,河南固始465200

出　　处：《动物医学进展》2010年第12期64-68,共5页Progress In Veterinary Medicine

摘　　要：参照GenBank收录的产毒素多杀性巴氏杆菌toxA基因序列(AF240778),设计一对引物,从猪源D型产毒素多杀性巴氏杆菌中扩增出toxA编码区3 858 bp的片段,将其克隆至原核表达载体PET-32a,转入大肠埃希菌BL21(DE3)中进行融合表达多杀性巴氏杆菌毒素。SDS-PAGE和Western blot分析证实,该表达产物以包涵体形式存在,大小约175 ku。动物试验结果表明,重组蛋白具有良好的免疫原性、反应原性及一定的生物学活性。通过间接ELISA方法检测抗毒素的猪阳性血清,表达重组蛋白较天然蛋白具有更高的特异性,为多杀性巴氏杆菌毒素进一步应用奠定了基础。According to published toxA gene sequence of Pasteurella multocida,a pair of primers were designed and synthesized,a 3 858 bp fragment was amplified from toxigenic Pasteurella multocida serotype D by PCR.The acquired PCR products were inserted into prokaryotic expression PET-32a（＋）,and transformed to E.coli BL21（DE3） for expression induced by isopropyl-β-D-thiogalactopyranoside（IPTG）.The expressed recombinant proteins were detected by SDS-PAGE and Western-blot.The results indicated that the protein had good immunological activity.The specificity of the recombinant protein was higher than that of the native protein.This study provides useful data for further use of the PMD（Pasteurella multocida serotype D）.

关 键 词：猪源D-型产毒素多杀巴氏杆菌 毒素基因 原核表达 

分 类 号：S852.612[农业科学—基础兽医学] Q786[农业科学—兽医学]