B型副鸡嗜血杆菌血凝素基因的克隆表达及生物活性鉴定  

Cloning and Expression of HA Gene of Serum-type B of Haemophilus paragallinarum in E.coli and Study on Biological Activity of Recombinant Protein

作　　者：王宏俊[1] 张培君[2] 龚玉梅[2] 杨汉春[1] 

机构地区：[1]中国农业大学动物医学院,北京100094 [2]北京市农林科学院畜牧兽医研究所,北京100097

出　　处：《畜牧兽医学报》2007年第1期101-104,共4页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：863项目资助(2003AA241120);北京市科技新星项目(2005B35)

摘　　要：血凝素是副鸡嗜血杆菌(Haemophilus paragallinarum,Hpg)的主要抗原成分之一,鸡只对Hpg的免疫力与血凝素抗体滴度成正相关。根据GenBank上已发表的B型Hpg Dalian株的血凝素基因序列(AY622378),设计合成了1对特异扩增Hpg血凝素基因的引物。以大连分离株Hpg中提取的细菌DNA为模板,利用PCR扩增出了1 038 bp的血凝素基因,将其克隆到pET-32a载体上,构建了pET-HA原核表达质粒并在BL21大肠杆菌中过量表达。血凝试验显示纯化的重组蛋白具有血凝活性;Western试验证明该重组蛋白可以被B型Hpg特异性鸡血清所识别。本研究在国内首次对Hpg的血凝素基因进行克隆表达,并分析了重组蛋白的血凝活性。There is a close correlation between HI titre of Haemophilus paragallinarum （Hpg） HA and protection from infectious coryza. According to the published hemagglutinin antigen （HA） gene sequence of Dalian strain of Hpg, a pair of primers was designed and synthesized for amplifing Hpg＇s major protecting antigen （HA gene）. A DNA fragment about 1 038 bp was obtained through PCR method from Dalian strain of Hpg. The HA gene was cloned into the expression vector pET-32a and overexpressed in E. coli BL21 （DE3）. Hemagglutination test indicated that the purified recombinant protein possess hemagglutinating activity. Western blot test showed that the protein was recognized by the chicken antibody against the Hpg of serotype B. This is first study in China to express HA gene and analysis the recombinant protein.

关 键 词：副鸡嗜血杆菌 血凝素 原核表达 

分 类 号：S852.612[农业科学—基础兽医学]