Improved recovery of rabies virus from cloned cDNA using a vaccinia virus-free reverse genetics system

Microbiol Immunol. 2003;47(8):613-7. doi: 10.1111/j.1348-0421.2003.tb03424.x.


To improve efficiency of recovery of rabies virus from cloned cDNA, we established a BHK cell clone that stably expresses T7 RNA polymerase, which we named BHK/T7-9. We also constructed new helper plasmids for expression of nucleoprotein and RNA polymerase of the RC-HL strain using the pTM1 plasmid vector, which makes the T7 RNA polymerase-transcripts from the plasmid cap-independent for translation. After co-transfection of these helper plasmids and the previously constructed full-length genome plasmid of the RC-HL strain to BHK/T7-9 cells, recombinant rabies virus was efficiently recovered from the cloned cDNA.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line
  • Cloning, Molecular / methods
  • DNA, Complementary / genetics*
  • DNA, Complementary / isolation & purification
  • Gene Expression Regulation, Viral
  • Genome, Viral
  • Plasmids
  • Rabies / virology*
  • Rabies virus / genetics
  • Rabies virus / isolation & purification*
  • Rabies virus / pathogenicity
  • Recombination, Genetic


  • DNA, Complementary