串联表达马立克氏病病毒糖蛋白B主要抗原决定簇基因的重组鸡痘病毒的构建

马立克氏病(Marek's disease,MD)是由马立克氏病病毒(Marek's disease virus,MDV)引起的一种具有高度传染性、淋巴组织增生性疾病,病理上以外周神经、性腺、虹膜、各种脏器和皮肤发生单核细胞浸润为特征[1].MD是世界上最重要的家禽传染病之一,是造成世界养禽业重大经济损失的一个主要原因.20世纪70年代早期MD疫苗的发现才使MD得到控制[2].

Construction of Recombinant Fowl Poxvirus Tandem Expressing the Major Antigen Cluster Gene of Glycoprotein B of Marek's Disease Virus

According to the data concerned and with the aids of DNA star software,the 250-460 amino acids of the glycoprotein B of Marek's disease virus GA strain was selected as target fragment for tandem expression.Two pairs of primers were designed for amplifying the upstream fragment gB(U) and the downstream fragment gB(L/D).And an ATG GCG was added to the 5' terminus of gB(U),a TAA was added to the 3' terminus of gB(L/D) and a linker of nine amino acids was designed at the 5' terminus of gB(L/D).Thus a complete ORF formed when gB(U) contacted with gB(L/D).PCR products were cloned by general molecular clone method.A transfer plasmid vector pEFMDgB(U/L/D) was constructed and then cotransfected with fowl poxvirus in CEF by calcium phosphate method and the recombinant virus was selected with MPA and its specificity was identified positive in infected CEF.The result showed that the tandem expressing products have immunocompetence,and this will bring a prospect for developing a candidate MD vaccine.