title: Improved recovery of rabies virus from cloned cDNA using a vaccinia virus-free reverse genetics system.
abstract: 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.
authors.affiliation: Laboratory of Zoonotic Diseases, Division of Veterinary Medicine, Faculty of Agriculture, Gifu University, Gifu, Gifu 501-1193, Japan.
authors.forename: Naoto
authors.initials: N
authors.lastname: Ito
authors.forename: Mutsuyo
authors.initials: M
authors.lastname: Takayama-Ito
authors.forename: Kentaro
authors.initials: K
authors.lastname: Yamada
authors.forename: Junji
authors.initials: J
authors.lastname: Hosokawa
authors.forename: Makoto
authors.initials: M
authors.lastname: Sugiyama
authors.forename: Nobuyuki
authors.initials: N
authors.lastname: Minamoto
mesh_terms: D002460:Cell Line; D003001:Cloning, Molecular / Q000379:methods; D018076:DNA, Complementary / Q000235:genetics* / Q000302:isolation & purification; D015967:Gene Expression Regulation, Viral; D016679:Genome, Viral; D010957:Plasmids; D011818:Rabies / Q000821:virology*; D011820:Rabies virus / Q000235:genetics / Q000302:isolation & purification* / Q000472:pathogenicity; D011995:Recombination, Genetic
chemical_list: D018076:DNA, Complementary