摘要
以鹅源H5亚型禽流感病毒 (AIV)基因组为模板 ,用RT PCR扩增血凝素 (Hemaggluti nin ,HA)基因 ,克隆入鸡痘病毒表达载体pFG1 1 75 ,转染鸡痘病毒感染的鸡胚成纤维细胞 ,通过蓝斑筛选和间接免疫荧光检测 ,获得表达HA基因的重组鸡痘病毒 (Recombinantfowlpoxvir us,rFPV HA)。rFPV HA经鸡胚成纤维细胞连续传 1 5代后 ,报告基因LacZ和HA基因可稳定表达。用 1 0 3PFU和 1 0 5PFU的rFPV HA免疫无特定病原体的 (Specificpathogenfree,SPF)鸡 ,免疫后 2 2d血凝抑制 (Hemagglutinininhibition ,HI)抗体监测阳性率分别为 0 %和 2 0 % ,但均抵御了H5亚型毒株的致死性攻击 ,保护率为 1 0 0 %。结果表明 ,构建了表达HA基因的重组鸡痘病毒 ,该重组病毒具有良好遗传稳定性 ,免疫鸡可提供完全保护 ,显示出了一定的应用前景。
Construction, genetic stability and protective efficacy of a recombinant fowlpox virus expressing hemagglutinin(HA) gene (rFPV-HA) of H5 subtype avian influenza virus were conducted. The results indicated that the recombinant fowlpox virus was obtained and purified by blue plaque selection. HA expressed by rFPV-HA was identified by indirect immunofluorescence array. The rFPV-HA could express reporter gene LacZ and HA gene after fifteen successive passages in chicken embryo fibroblast. The rFPV-HA induced completely protection of chickens against lethal challenge with homologous avian influenza virus. The results show that rFPV-HA has good genetic stability and antigenicity and should be a useful tool in HPAI control program by preventing illness and death in chickens when the disease spreads widely.
出处
《微生物学报》
CAS
CSCD
北大核心
2003年第6期722-727,共6页
Acta Microbiologica Sinica
基金
国家"8 63计划"(2 0 0 1AA2 1 3 0 41 )
农业部农办牧 2 0 0 1 83项目~~
关键词
H5亚型
禽流感
血凝素基因
重组鸡痘病毒
遗传稳定性
免疫效力
H5 subtype, Avian influenza, Hemagglutinin gene, Recombinant fowlpox virus, Genetic stability, Protective efficacy
