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NP and l proteins of lymphocytic choriomeningitis virus (LCMV) are sufficient for efficient transcription and replication of LCMV genomic RNA analogs

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Overview

authors

  • Lee, K. J.
  • Novella, I. S.
  • Teng, M. N.
  • Oldstone, Michael
  • de la Torre, Juan

publication date

  • April 2000

journal

  • Journal of Virology  Journal

abstract

  • The genome of lymphocytic choriomeningitis virus (LCMV) consists of two negative-sense single-stranded RNA segments, designated L and S. Both segments contain two viral genes in an ambisense coding strategy, with the genes being separated by an intergenic region (IGR). We have developed a reverse genetic system that allows the investigation of cis-acting signals and trans-acting factors involved in transcription and replication of LCMV. To this end, we constructed an LCMV S minigenome consisting of a negative-sense copy of the chloramphenicol acetyltransferase (CAT) reporter gene flanked upstream by the S 5' untranslated region (UTR) and IGR and downstream by the S 3' UTR. CAT expression was detected in LCMV-infected cells transfected with the minigenome RNA. Intracellular coexpression of the LCMV minigenome and LCMV L and NP proteins supplied from cotransfected plasmids driven by the T7 RNA polymerase provided by the recombinant vaccinia virus vTF7-3 resulted in high levels of CAT activity and synthesis of subgenomic CAT mRNA and antiminigenome RNA species. Thus, L and NP represent the minimal viral trans-acting factors required for efficient RNA synthesis mediated by LCMV polymerase.

subject areas

  • Animals
  • Cell Line
  • Cells, Cultured
  • Chloramphenicol O-Acetyltransferase
  • Cricetinae
  • DNA-Directed RNA Polymerases
  • Genome, Viral
  • Helper Viruses
  • Immunoblotting
  • Lymphocytic choriomeningitis virus
  • Nucleocapsid Proteins
  • Nucleoproteins
  • Plasmids
  • RNA, Viral
  • Transcription, Genetic
  • Transfection
  • Viral Core Proteins
  • Viral Proteins
  • Virus Replication
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Identity

International Standard Serial Number (ISSN)

  • 0022-538X

Digital Object Identifier (DOI)

  • 10.1128/jvi.74.8.3470-3477.2000

PubMed ID

  • 10729120
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Additional Document Info

start page

  • 3470

end page

  • 3477

volume

  • 74

issue

  • 8

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