摘要
本试验构建了H9亚型禽流感病毒HA基因禽痘病毒转移载体,经转染、蓝斑克隆、筛选和纯化,获得了遗传性状稳定的HA基因重组禽痘病毒。提取感染重组病毒的鸡胚成纤维细胞(CEF)的DNA进行PCR扩增,获得1.7kb的携带有外源目的基因片段。收集纯化的重组病毒CEF细胞,用H9亚型禽流感病毒多克隆血清作一抗,碱性磷酸酶标记的鸡IgG为二抗进行Western blot检测,结果表明重组痘病毒能在体外的CEF细胞表达HA糖蛋白。
In this study, to produce a kind of vaccine including cross-protection in chickens, recombinant fowlpox virus （rFPV） expressing HA9 gene of AIV was constructed. After construction of transfer vector, it was transfected on CEF cells with FPV. Following clonning and purification of blue plaque, the pure HA-fowlpox virus recombinant was obtained, and identified by restriction enzyme analysis and PCR. The result showed that the recombinant fowlpox virus contained HA gene of AIV and had the stable genetic properties. The result of Western-blot showed that the recombinant fowlpox virus expressed the hemagglutinin in glycoprotein of AIV efficiently.
出处
《中国动物检疫》
CAS
2009年第3期31-34,共4页
China Animal Health Inspection
