1998
DOI: 10.1128/jvi.72.9.7270-7279.1998
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trans -Complementation of Flavivirus RNA Polymerase Gene NS5 by Using Kunjin Virus Replicon-Expressing BHK Cells

Abstract: A BHK cell line persistently expressing a Kunjin (KUN) virus replicon RNA (repBHK, similar to our recently described ME/76Neo BHK cell line [A. A. Khromykh and E. G. Westaway, J. Virol. 71:1497–1505, 1997]) was used for rescue and propagation of KUN viruses defective in the RNA polymerase gene (NS5). A new infectious full-length KUN virus cDNA clone, FLSDX, prepared from our previously described cDNA clone pAKUN (A. A. Khromykh and E. G. Westaway, J. Virol. 68:4580–4588, 1994) and possessing ∼105-fold higher s… Show more

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Cited by 135 publications
(60 citation statements)
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References 35 publications
(53 reference statements)
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“…The multiple points of regulation present a formidable theoretical barrier to extensive deletions or insertions in polyproteins. Several groups working with positive-stranded RNA viruses have reported deletions of portions of protein domains or the generation of viruses whose extensive defects in replication require rescue by replicons or helper viruses (16,26,29,32,34,38,39,44,48). However, for animal positive-stranded RNA viruses, we have identified no published reports of autonomously replicating mutant viruses with deletions of complete mature protein domains within polyproteins.…”
mentioning
confidence: 87%
“…The multiple points of regulation present a formidable theoretical barrier to extensive deletions or insertions in polyproteins. Several groups working with positive-stranded RNA viruses have reported deletions of portions of protein domains or the generation of viruses whose extensive defects in replication require rescue by replicons or helper viruses (16,26,29,32,34,38,39,44,48). However, for animal positive-stranded RNA viruses, we have identified no published reports of autonomously replicating mutant viruses with deletions of complete mature protein domains within polyproteins.…”
mentioning
confidence: 87%
“…While ideal, trans-complementation using protein expression has not been successful for coronaviruses. In fact, efficient complementation for other positive-strand RNA viruses has required the development of replicon systems to facilitate viral replication complex formation (Appel et al, 2005;Grassmann et al, 2001;Khromykh et al, 1998;Lindenbach and Rice, 1997;Liu et al, 2002). Despite these limitations, this is the first report of detailed mutagenesis of an MHV gene 1 nsp-coding region within the context of a virus.…”
Section: Importance Of the Nsp1 Carboxy-terminal Domainmentioning
confidence: 99%
“…A third approach involves the use of medium-or low-copy-number plasmids to stabilize full-length flavivirus cDNAs. This strategy has been employed with different strains of DENV2 cDNAs (23,36,51,71), West Nile virus cDNA (55,66), Kunjin virus cDNA (34,35), and TBEV cDNA (43). Two other modified methods have been derived from this strategy to facilitate the construction of flavivirus cDNAs.…”
mentioning
confidence: 99%