TY - JOUR
T1 - An improved method for recovering rabies virus from cloned cDNA
AU - Inoue, Ken Ichi
AU - Shoji, Youko
AU - Kurane, Ichiro
AU - Iijima, Toshio
AU - Sakai, Takeo
AU - Morimoto, Kinjiro
N1 - Funding Information:
We thank Karl-Klaus Conzelmann (Max von Pettenkofer Institute and Gene Center, Germany) for kindly supplying the BSR-T7/5 cells. This work was supported by grants from the Research on Emerging and Re-emerging Infectious Diseases, Ministry of Health, Labor and Welfare, Japan.
PY - 2003/2
Y1 - 2003/2
N2 - 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.
AB - 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.
KW - Cytomegalovirus promotor
KW - HEP-Flury strain
KW - Rabies virus
KW - Reverse genetics
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U2 - 10.1016/S0166-0934(02)00249-5
DO - 10.1016/S0166-0934(02)00249-5
M3 - Article
C2 - 12505638
AN - SCOPUS:0037291509
VL - 107
SP - 229
EP - 236
JO - Journal of Virological Methods
JF - Journal of Virological Methods
SN - 0166-0934
IS - 2
ER -
