麻鸡副黏病毒ZH-1株HN基因真核表达载体的构建及其免疫试验

Construction and administration of nucleic acid vaccine expressing HN gene of paramyxovirus ZH-1 strain from spotted chickens

应用RT-PCR方法从麻鸡副黏病毒ZH-1株中扩增HN基因并克隆入pGEM-T载体,经序列测定、分析,结果显示ZH-1株HN基因与NDV国家标准强毒F48E9株的核苷酸、氨基酸序列同源性均达到92%。将HN基因克隆入真核表达载体pCI-neo,构建真核表达质粒pC-IHN,转染Vero细胞,能在Vero细胞中表达,利用pCI-HN质粒进行动物试验,结果表明,雏鸡免疫14d后在体内可检测到特异性抗体,pC-IHN质粒二次免疫雏鸡后,对NDV强毒的攻毒保护率为67%,这一结果提示,利用HN基因构建的核酸疫苗具有重要的开发应用价值。

In this study,Hemagglutinin-Neuramidinase glycoprotein（HN） genes of paramyxovirus ZH-1 strain isolated from spotted chickens were amplified by reverse transcriptase polymerase chain reaction（RT-PCR） and cloned into pGEM-T vector.The sequence analyses of HN genes indicated that the homology of nucleotide acid and amino acid sequences between ZH-1 strain and F48E9 strain,standard virulent strain of NDV were more then 92%.The HN genes were cloned into the eukaryotic expression vector PCI-neo.And the recombinant plasmid pCI-HN was transferred into vero cells to express HN protein.The chickens immunized by the pCI-HN displayed specific antibody at 14 days post inoculation.After second immunization,protective rate of the chicken challenged by NDV standard virulent infection was 67%.The results suggest that the nucleic acid vaccine expressing HN gene for protecting spotted chicken was valuable for further development.