摘要
以鸡传染性喉气管炎病毒(ILTV)LJS09株基因组为模板,应用PCR方法扩增gG基因的主要抗原性片段,并克隆至原核表达载体pET-30a(+)构建pET-30a-gG-gG串联质粒,转化Rosetta(DE3),经IPTG诱导表达及纯化,获得His-gG-gG重组蛋白,免疫新西兰大白兔制备抗血清。用ELISA检测抗体效价为1∶104。Western-blot分析结果表明,制备的家兔抗-ILTV gG抗体能与纯化的gG蛋白发生特异性反应,且能检测到内源性gG蛋白。间接免疫荧光试验结果表明,制备的家兔抗ILTV-LJS09gG蛋白的多克隆抗体分别能与ILTV LJS09、HLJ0507、MDJ0442、SD0203、Zhj1298株反应,特异性良好。
Major antigenic fragment of gG gene of infectious laryngotracheitis virus（ILTV） was ampli-fied from ILTV-LJS09 genome by PCR. The prokaryotic expression vector pET-30a-gG-gG was construc-ted by cloning the fragment into the pET-30a vector. The His-gG-gG recombinant protein was obtained in Rosetta（DE3） induced with IPTG,and used to immunize rabbits. The titer of the antibodies was 1 ： 10 in ELISA. The obtained polyclonal antibodies were specific to ILTV-LJS09 antigen in a Western-blot test. In-direct immunofluorescence assay showed that anti ILTV-LJS09 gG protein antibodies could specifically respond to ILTV-LJS09, HLJ0507 ,MDJ0442 ,SD0203 ,and Zhj1298 strains.
出处
《中国兽医科学》
CAS
CSCD
北大核心
2014年第2期165-170,共6页
Chinese Veterinary Science
基金
中央级公益性科研院所基本科研业务费项目(0302012009)
关键词
传染性喉气管炎病毒
gG蛋白
原核表达
多克隆抗体
间接免疫荧光试验
infectious laryngotracheitis virus （ILTV）
gG protein
prokaryotic expression
polyclonalantibody
indirect immunofluorescence assay（IFA）
