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Published on: November 1, 2011
Development of a reverse genetics system for the aG strain of rabies virus in China
Jiao Wenqiang1, Xiangping Yin, Xi Lan
1State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Epizootic Disease of Grazing Animal of Ministry of Agriculture, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Science (LVRI, CAAS), Xujia ping1, Yanchang bu, Lanzhou, 730046, Gansu, China.
Abstract:
The aG rabies virus strain has been attenuated through multiple passages in cells and is now used as a vaccine strain in China. We attempted to develop a reverse genetics system using the aG strain. Recombinant full-length genomic cDNA was flanked by a hammerhead ribozyme and the hepatitis delta virus ribozyme. Three helper plasmids encoding the nucleoprotein, the phosphoprotein, and the large protein were produced and introduced together with a plasmid containing the full-length aG viral genome into BHK-21 cells by transfection. Recombinant virus was successfully recovered from the cloned cDNA under the control of a CMV promoter driven by RNA polymerase II. The recombinant virus was confirmed by RT-PCR, and the titer of the recombinant virus was 6.2 log LD50.
