An improved method for recovering rabies virus from cloned cDNA.
A new system for recovery of rabies virus from cDNA plasmid, the transcription of which was driven by cellular RNA polymerase II, was developed. The plasmid contains full-length viral cDNA flanked by hammerhead ribozyme and hepatitis delta ribozyme sequences, arranged downstream of the cytomegalovirus (CMV) promotor. Transfection with the full-length cDNA plasmid together with helper plasmids encoding viral N, P, and L proteins without supply of T7 RNA polymerase produced a recombinant rabies virus in several cell lines. The efficiency of recovery between the conventional T7 promotor system and the new CMV promotor system was compared using these plasmid constructs. The newly established system is applicable to various cell lines and allows rapid and efficient generation of recombinant rabies virus.
Inoue
Ken ichi
Ki
Department of Virology I, National Institute of Infectious Diseases, Toyama 1-23-1, Shinjuku-ku, 162-8640, Tokyo, Japan.
Shoji
Youko
Y
Kurane
Ichiro
I
Iijima
Toshio
T
Sakai
Takeo
T
Morimoto
Kinjiro
K
D000818:Animals; D002460:Cell Line; D003001:Cloning, Molecular / Q000379:methods*; D006224:Cricetinae; D003587:Cytomegalovirus; D018076:DNA, Complementary / Q000235:genetics*; D006801:Humans; D051379:Mice; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011820:Rabies virus / Q000235:genetics / Q000302:isolation & purification*; D011995:Recombination, Genetic; D014158:Transcription, Genetic; D014162:Transfection
D018076:DNA, Complementary