新城疫热稳定性天然弱毒B95株核酸疫苗的构建

Construction and vaccination test of nucleotide vaccine of F glycoprotein gene from natural attenuated strain of Newcastle disease virus

将含有新城疫热稳定性天然弱毒B95株F基因的重组克隆质粒 pGEM F用NotⅠ单酶切 ,电泳回收纯化目的片段 ,与同样用NotⅠ单酶切并经去磷酸化处理的真核表达载体 pcDNA3.1连接 ,构建了新城疫核酸疫苗 ,并用所构建的新城疫核酸疫苗与载基因阳离子脂质体结合进行了免疫试验 ,通过ELISA、淋巴细胞转化试验检测了核酸疫苗在鸡体内的表达。结果显示 ,表达产物作为抗原物质刺激机体产生了特异性应答反应 ,引起了机体特异性的体液免疫和细胞免疫 ,对新城疫强毒攻击的保护率为 75 %。

The cDNA fragment encoding F glycoprotein of B95, a natural attenuated strain of New-(castle) disease virus, was obtained by digesting recombinant plasmid pGEM-F with NotⅠ and being purifying with the agar-gel fraction method. This cDNA fragment was then cloned into eukaryotic expression vector pcDNA3.1 to construct nucleotide vaccine, pcDNA-F. The pcDNA-F was then used to vaccinate chicken with positive liposome as adjuvant. The special humoral and cell immunity were detected by ELISA and lymphocyte transformation test. The special responses were produced in vaccinated chickens and 75% (protection) was obtained when challenged with virulent NDV F48E9. This study has laid a foundation for (further) studying recombinant vaccine against NDV.