HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by McCormick, C. J. Articles by Harris, M. Search for Related Content PubMed PubMed Citation Articles by McCormick, C. J. Articles by Harris, M. Agricola Articles by McCormick, C. J. Articles by Harris, M.

Tagging of NS5A expressed from a functional hepatitis C virus replicon

Christopher J. McCormick^,

Sophie Maucourant

Institute of Molecular and Cellular Biology, Faculty of Biological Sciences, University of Leeds, Leeds LS2 9JT, UK

Correspondence
Mark Harris
m.harris{at}leeds.ac.uk

Knowledge of how hepatitis C virus (HCV) proteins associate with components of the host cell to form a functional replication complex is still limited. To address this issue, HCV replicon constructs were generated where either green fluorescent protein (GFP) or the Propionibacterium shermanii transcarboxylase domain (PSTCD) was introduced into the NS5A coding region. Insertion of both GFP and PSTCD was tolerated well, allowing formation of stable replicon-containing cell lines that contained viral protein and transcript levels that were comparable to those of an unmodified parental replicon. Cell lines generated from the GFP-tagged NS5A replicon allowed live-cell visualization of the location of NS5A. Cell lines generated from the PSTCD-tagged replicons allowed rapid and efficient precipitation of the PSTCD-tagged NS5A, as well as other HCV non-structural proteins, using streptavidin-coated magnetic beads. Both replicons represent useful tools that offer different but complementary ways of examining replication-complex formation in cells.

These authors contributed equally to this work.

Present address: Molecular Microbiology and Infection, University of Southampton, Southampton General Hospital, Southampton SO16 6YD, UK.

This article has been cited by other articles:

				M. Hughes, S. Griffin, and M. Harris Domain III of NS5A contributes to both RNA replication and assembly of hepatitis C virus particles J. Gen. Virol., June 1, 2009; 90(6): 1329 - 1334. [Abstract] [Full Text] [PDF]

				J. Mankouri, A. Milward, K. R. Pryde, L. Warter, A. Martin, and M. Harris A comparative cell biological analysis reveals only limited functional homology between the NS5A proteins of hepatitis C virus and GB virus B J. Gen. Virol., August 1, 2008; 89(8): 1911 - 1920. [Abstract] [Full Text] [PDF]

				T. L. Tellinghuisen, K. L. Foss, J. C. Treadaway, and C. M. Rice Identification of Residues Required for RNA Replication in Domains II and III of the Hepatitis C Virus NS5A Protein J. Virol., February 1, 2008; 82(3): 1073 - 1083. [Abstract] [Full Text] [PDF]

				T. L. Tellinghuisen, M. J. Evans, T. von Hahn, S. You, and C. M. Rice Studying Hepatitis C Virus: Making the Best of a Bad Virus J. Virol., September 1, 2007; 81(17): 8853 - 8867. [Full Text] [PDF]

				D. M. Jones, S. N. Gretton, J. McLauchlan, and P. Targett-Adams Mobility analysis of an NS5A-GFP fusion protein in cells actively replicating hepatitis C virus subgenomic RNA J. Gen. Virol., February 1, 2007; 88(2): 470 - 475. [Abstract] [Full Text] [PDF]