Infectious rabies viruses from cloned cDNA.
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TLDR
The generation of infectious rabies virus (RV), a non‐segmented negative‐stranded RNA virus of the Rhabdoviridae family, entirely from cloned cDNA is described, and the possibility of manipulating the RV genome by recombinant DNA techniques using the described procedure greatly facilitates the investigation of RV genetics, virus‐host interactions and rabies pathogenesis.Abstract:
The generation of infectious rabies virus (RV), a non-segmented negative-stranded RNA virus of the Rhabdoviridae family, entirely from cloned cDNA is described. Simultaneous intracellular expression of genetically marked full-length RV antigenome-like T7 RNA polymerase transcripts and RV N, P and L proteins from transfected plasmids resulted in formation of transcriptionally active nucleocapsids and subsequent assembly and budding of infectious rabies virions. In addition to authentic RV, two novel infectious RVs characterized by predicted transcription patterns were recovered from modified cDNA. Deletion of the entire non-translated pseudogene region, which is conserved in all naturally occurring RVs, did not impair propagation of the resulting virus in cell culture. This indicates that non-essential genetic material might be present in the genomes of non-segmented RNA viruses. The introduction of a functional extra cistron border into the genome of another virus resulted in the transcription of an additional polyadenylated mRNA containing pseudogene sequences. The possibility of manipulating the RV genome by recombinant DNA techniques using the described procedure--potentially applicable also for other negative-stranded viruses--greatly facilitates the investigation of RV genetics, virus-host interactions and rabies pathogenesis and provides a tool for the design of new generations of live vaccines.read more
Citations
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Journal ArticleDOI
Generation of influenza A viruses entirely from cloned cDNAs
Gabriele Neumann,Tokiko Watanabe,Hiroshi Ito,Shinji Watanabe,Hideo Goto,Peng Gao,Mark A. Hughes,Daniel R. Perez,Ruben O. Donis,Erich Hoffmann,Gerd Hobom,Yoshihiro Kawaoka +11 more
TL;DR: A new reverse-genetics system that allows one to efficiently generate influenza A viruses entirely from cloned cDNAs is described, which should be useful in viral mutagenesis studies and in the production of vaccines and gene therapy vectors.
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Generation of bovine respiratory syncytial virus (BRSV) from cDNA: BRSV NS2 is not essential for virus replication in tissue culture, and the human RSV leader region acts as a functional BRSV genome promoter.
TL;DR: The successful recovery of a BRSV mutant lacking the complete NS2 gene, which encodes a nonstructural protein of unknown function is reported, demonstrating that NS2 is not essential for virus replication in cell culture.
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Rescue of Influenza A Virus from Recombinant DNA
Ervin Fodor,Louise J. Devenish,Othmar G. Engelhardt,Peter Palese,George G. Brownlee,Adolfo García-Sastre +5 more
TL;DR: The rescued influenza A virus is rescued by transfection of 12 plasmids into Vero cells by plasmid-based reverse genetics technique, which facilitates the generation of recombinant influenza viruses containing specific mutations in their genes.
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Rescue of measles viruses from cloned DNA.
Frank Radecke,Pius Spielhofer,Henriette Schneider,Karin Kaelin,M. Huber,C. Dötsch,G. Christiansen,Martin A. Billeter +7 more
TL;DR: This system, in principle, should be applicable to the rescue of any member of the large virus order Mononegavirales, i.e. viruses with a nonsegmented negative‐strand RNA genome.
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Recombinant vesicular stomatitis viruses from DNA
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References
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