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 ...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.展开更多
文摘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.
