麻鸡副黏病毒ZH-1株F基因核酸疫苗的构建及其免疫效果

Construction and immunogenicity of nucleic acid vaccine expressing F gene of paramyxovirus ZH-1 strain from spotted chicken

应用RT-PCR方法从麻鸡副黏病毒ZH-1株中扩增F基因并克隆入pGEM○R-T载体,经序列测定、分析,结果显示ZH-1株F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99%。随后将F基因克隆入真核表达载体pCI-neo,构建真核表达质粒pCI-F,pCI-F体外转染Vero细胞,能在Vero细胞中表达,利用pCI-F质粒进行动物试验,结果表明雏鸡免疫14 d后在体内可检测到特异性抗体,pCI-F质粒二次免疫雏鸡后,对NDV强毒的攻毒保护率为73%,这一结果提示利用F基因构建的核酸疫苗具有重要的开发前景。

In this study, fusion glycoprotein（F）gene of paramyxovirus ZH-1 strain isolated from spotted chicken was amplified by reverse transcriptase ploymerase chain reaction（RT-PCR）and cloned into pGEM- T vector. The sequence analyses of F gene indicated that the homology of nucleotide acid sequences and amino acid sequences between ZH-1 strain and F48E9 strain, standard virulent strain of NDV was 99 %. Then, F gene was cloned into the eukaryotie expression vector pCI neo. The recombinant plasmid pCI-F was transferred into vero cells to express F protein. The chickens immuned by displayed specific antibody at 14 d. After second immunization, protectve rate of the chicken challenged by NDV standard virulent infection was 73%. The result suggestted that it is worth to construct nucleic acid vaccine expressing F gene.