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Rescue of the prototypic arenavirus LCMV entirely from plasmid

Academic Article
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Overview

authors

  • Sanchez, A. B.
  • de la Torre, Juan

publication date

  • July 2006

journal

  • Virology  Journal

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.

subject areas

  • Animals
  • Base Sequence
  • Cell Line
  • Kidney
  • Lymphocytic choriomeningitis virus
  • Plasmids
  • Transfection
  • Virus Replication
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Research

keywords

  • Arenavirus
  • lymphocytic choriomeningitis virus (LCMV)
  • rescue
  • reverse genetics
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Identity

International Standard Serial Number (ISSN)

  • 0042-6822

Digital Object Identifier (DOI)

  • 10.1016/j.virol.2006.01.012

PubMed ID

  • 16476461
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Additional Document Info

start page

  • 370

end page

  • 380

volume

  • 350

issue

  • 2

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