Tagging of NS5A expressed from a functional hepatitis C virus replicon
Tagging of NS5A expressed from a functional hepatitis C virus replicon
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.
635-640
McCormick, C.J.
0fce14bf-2f67-4d08-991f-114dd1e7f0bd
Maucourant, S.
a332ba09-a25c-4fa9-b179-ddb04146bbb4
Griffin, S.D.J.
707c785a-4085-4df0-ab05-0f16f596b64d
Rowlands, David J.
2de54fdc-5f4f-40f3-b259-c8fb966a782a
Harris, M.
b0e2d8cb-44f8-47a5-8e1b-6ca9c04fe776
2006
McCormick, C.J.
0fce14bf-2f67-4d08-991f-114dd1e7f0bd
Maucourant, S.
a332ba09-a25c-4fa9-b179-ddb04146bbb4
Griffin, S.D.J.
707c785a-4085-4df0-ab05-0f16f596b64d
Rowlands, David J.
2de54fdc-5f4f-40f3-b259-c8fb966a782a
Harris, M.
b0e2d8cb-44f8-47a5-8e1b-6ca9c04fe776
McCormick, C.J., Maucourant, S., Griffin, S.D.J., Rowlands, David J. and Harris, M.
(2006)
Tagging of NS5A expressed from a functional hepatitis C virus replicon.
The Journal of General Virology, 87 (3), .
(doi:10.1099/vir.0.81553-0).
Abstract
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.
This record has no associated files available for download.
More information
Published date: 2006
Identifiers
Local EPrints ID: 27264
URI: http://eprints.soton.ac.uk/id/eprint/27264
PURE UUID: b4281fce-92ae-400e-9bdf-9a8d05a06a77
Catalogue record
Date deposited: 26 Apr 2006
Last modified: 16 Mar 2024 03:47
Export record
Altmetrics
Contributors
Author:
S. Maucourant
Author:
S.D.J. Griffin
Author:
David J. Rowlands
Author:
M. Harris
Download statistics
Downloads from ePrints over the past year. Other digital versions may also be available to download e.g. from the publisher's website.
View more statistics