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  

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

作　　者：钱莺娟[1] 张雪莲[1] 陈德胜[1] 陈溥言[1] 

机构地区：[1]南京农业大学农业部动物疫病诊断和免疫重点开放实验室,江苏南京210095

出　　处：《病毒学报》2004年第2期182-185,共4页Chinese Journal of Virology

基　　金：江苏省兽用生物制品工程中心开放项目资助

摘　　要：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.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.

关 键 词：马立克氏病 病毒 抗原决定簇 串联表达 糖蛋白B 鸡痘病毒 

分 类 号：S852.659.1[农业科学—基础兽医学] Q78[农业科学—兽医学]