Rescue of the prototypic Arenavirus LCMV entirely from plasmid.
Virology
; 350(2): 370-80, 2006 Jul 05.
Article
en En
| MEDLINE
| ID: mdl-16476461
ABSTRACT
We document a helper-independent reverse genetics system for rescuing infectious arenaviruses from cloned cDNAs. We constructed plasmids containing full-length cDNAs of the antigenomic (ag) L and S segments of the Armstrong (ARM) strain of the prototypic Arenavirus lymphocytic choriomeningitis virus (LCMV) flanked at their 5'- and 3'-termini by the T7 RNA polymerase (T7RP) promoter and ribozyme sequences, respectively. These plasmids directed intracellular synthesis of viral L and S ag RNA species in cells expressing plasmid-supplied T7RP. Co-expression of plasmid-supplied LCMV trans-acting factors, nucleoprotein (NP) and polymerase (L), resulted in replication and expression of L and S ag and genome RNA species, and generation of LCMV infectious progeny termed rT7/LCMV. The recombinant rT7/LCMV was unequivocally identified based on a genetic tag introduced in the recombinant S segment. In addition, rT7/LCMV exhibited growth and biological properties predicted for an ARM-like LCMV. To our knowledge, this is the first documented Arenavirus rescue, as well as of an ambisense negative strand (NS) RNA virus, entirely from cloned cDNAs. Our results extend the use of reverse genetic approaches for DNA-mediated virus rescue to all known virus families with NS RNA genome.
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Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Plásmidos
/
Virus de la Coriomeningitis Linfocítica
Tipo de estudio:
Prognostic_studies
Límite:
Animals
Idioma:
En
Revista:
Virology
Año:
2006
Tipo del documento:
Article
País de afiliación:
Estados Unidos