Author: Angelini, Megan M.; Akhlaghpour, Marzieh; Neuman, Benjamin W.; Buchmeier, Michael J.
Title: Severe Acute Respiratory Syndrome Coronavirus Nonstructural Proteins 3, 4, and 6 Induce Double-Membrane Vesicles Document date: 2013_8_13
ID: yl1qyh7j_20
Snippet: Plasmids and transfection. All plasmids used were created as previously described using a Gateway expression system (Invitrogen) (56) . Briefly, all constructs had a modified pCAGGs backbone containing a Woodchuck hepatitis virus posttranscriptional regulatory element (WPRE) and were C-terminally tagged with either an HA tag sequence followed by a tobacco etch virus (TEV) cleavage site and a biotinylation signal sequence (HA-Bio) or an HA tag seq.....
Document: Plasmids and transfection. All plasmids used were created as previously described using a Gateway expression system (Invitrogen) (56) . Briefly, all constructs had a modified pCAGGs backbone containing a Woodchuck hepatitis virus posttranscriptional regulatory element (WPRE) and were C-terminally tagged with either an HA tag sequence followed by a tobacco etch virus (TEV) cleavage site and a biotinylation signal sequence (HA-Bio) or an HA tag sequence followed by a 3Ï« FLAG tag sequence (HA-3Ï«FLAG). Transfections were conducted using Lipofectamine 2000 (Invitrogen) per the manufacturer's protocol.
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