关键词: 狂犬病病毒G基因, N基因, 内部核糖体进入位点序列, 重组腺病毒

Abstract: To construct adenoviral vector co-expressing G and N gene of rabies virus by utilizing an internal ribosome entry site (IRES) sequences to link the two genes, the G and N gene of rabies virus were obtained by reverse transcriptase-polymerase chain reaction (RT-PCR). The IRES\|N gene fragment was obtained by subcloning N gene into pIRES vector, then subcloned G and IRES-N into shuttle plasmid pAdTrack-CMV. The recombinant transfer vector linearized with Pme I digestion and pAdEasy-1 backbone vector was further cotransformed into the bacteria BJ5183 competent cells for homologous recombination. The resultant homologous recombinant plasmids were linearized with Pac I digestion and transfected into 293 cells to pack the adenovirus. Strong green fluorescence was observed by fluorescence microscope. The glycoprotein and nucleoprotein genes fragments of rabies virus could be detected in the recombinant adenovirus by RT-PCR. The results showed that the recombinant adenovirus containing G gene and N gene was successfully constructed and obtained, thus providing a basis for the research of recombinant replication\|defective adenovirus based rabies virus vaccine．

Key words: G gene of rabies virus, N gene, IRES, recombinant adenovirus