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Recombinant lymphocytic choriomeningitis virus expressing vesicular stomatitis virus glycoprotein

Academic Article
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Overview

authors

  • Pinschewer, D. D.
  • Perez, M.
  • Sanchez, A. B.
  • de la Torre, Juan

publication date

  • June 2003

journal

  • Proceedings of the National Academy of Sciences of the United States of America  Journal

abstract

  • A recombinant S segment RNA (Sr) of the prototypic arenavirus lymphocytic choriomeningitis virus (LCMV) where the glycoprotein of vesicular stomatitis virus (VSVG) was substituted for the glycoprotein of LCMV (LCMV-GP) was produced intracellularly from cDNA under the control of a polymerase I promoter. Coexpression of the LCMV proteins NP and L allowed expression of VSVG from Sr. Infection of transfected cells with WT LCMV (LCMVwt) resulted in reassortment of the L segment of LCMVwt with the Sr at low frequency. Isolation of recombinant LCMV (rLCMV) expressing VSVG (rLCMV/VSVG) was achieved by selection against LCMVwt by using a cell line deficient in the cellular protease S1P. This approach was based on the finding that processing of LCMV-GP by S1P was required for virus infectivity. Characterization of protein and RNA expression of rLCMV/VSVG in infected cells confirmed the expected virus genome organization. rLCMV/VSVG caused syncytium formation in cultured cells and grew to approximately 100-fold lower titers than WT virus but, like the parent virus, it persisted in neonatally infected mice without clinical signs of disease.

subject areas

  • Animals
  • Blotting, Northern
  • Blotting, Western
  • CHO Cells
  • Cercopithecus aethiops
  • Cricetinae
  • Genetic Engineering
  • Genetic Techniques
  • Lymphocytic choriomeningitis virus
  • Membrane Glycoproteins
  • Mice
  • Mice, Inbred BALB C
  • Microscopy, Fluorescence
  • Plasmids
  • RNA
  • Reverse Transcriptase Polymerase Chain Reaction
  • Time Factors
  • Transfection
  • Vero Cells
  • Viral Envelope Proteins
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Identity

PubMed Central ID

  • PMC164684

International Standard Serial Number (ISSN)

  • 0027-8424

Digital Object Identifier (DOI)

  • 10.1073/pnas.1332709100

PubMed ID

  • 12808132
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Additional Document Info

start page

  • 7895

end page

  • 7900

volume

  • 100

issue

  • 13

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