Author: Stewart, Meredith E.; Roy, Polly
Title: Structure-based identification of functional residues in the nucleoside-2'-O-methylase domain of Bluetongue virus VP4 capping enzyme Document date: 2015_2_24
ID: vzel6r43_6
Snippet: The coding region of S4 (GenBank: FJ969722), VP4, was amplified and ligated into expression vector pCAG-Puro (pCAG-VP4). BSR cells were transfected with the pCAG-VP4 plasmid using Lipofectamine2000 (Invitrogen) as per instruction. After transfection, cells with integrated copies of the expression vector were selected by the addition of 7.5 lg/ml of puromycin (Sigma). The surviving clones were tested for the expression level of VP4 by western immu.....
Document: The coding region of S4 (GenBank: FJ969722), VP4, was amplified and ligated into expression vector pCAG-Puro (pCAG-VP4). BSR cells were transfected with the pCAG-VP4 plasmid using Lipofectamine2000 (Invitrogen) as per instruction. After transfection, cells with integrated copies of the expression vector were selected by the addition of 7.5 lg/ml of puromycin (Sigma). The surviving clones were tested for the expression level of VP4 by western immunoblot and the ability to grow virus.
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