Development of a reverse genetics system for the aG strain of rabies virus in China

Development of a reverse genetics system for the aG strain of rabies virus in China 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 LD 50 . http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Archives of Virology Springer Journals

Development of a reverse genetics system for the aG strain of rabies virus in China

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Publisher
Springer Journals
Copyright
Copyright © 2014 by Springer-Verlag Wien
Subject
Biomedicine; Virology; Medical Microbiology; Infectious Diseases
ISSN
0304-8608
eISSN
1432-8798
D.O.I.
10.1007/s00705-013-1919-9
Publisher site
See Article on Publisher Site

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 LD 50 .

Journal

Archives of VirologySpringer Journals

Published: May 1, 2014

References

  • Development of a reverse genetics system for a human rabies virus vaccine strain employed in China
    Huang, Y; Tang, Q; Nadin-Davis, SA
  • Initiation of Sendai virus multiplication from transfected cDNA or RNA with negative or positive sense
    Kato, A; Sakai, Y; Shioda, T; Kondo, T; Nakanishi, M; Naqai, Y
  • Structure-function relationships of hammerhead ribozyme: from understanding to applications
    Sigurdsson, ST; Eckstein, F
  • Self-cleaving ribozyme of hepatitis delta virus RNA
    Been, MD; Wickham, GS
  • An improved method for recovering rabies virus from cloned cDNA
    Inoue, KI; Shoji, Y; Kurane, I; Iijima, T; Sakai, T; Morimoto, K
  • Glycoprotein gene relocation in rabies virus
    Wu, X; Rupprecht, CE

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