Journal:

Article Title: Hepatitis C Virus-Like Particle Budding: Role of the Core Protein and Importance of Its Asp 111

doi: 10.1128/JVI.77.18.10131-10138.2003

Figure Lengend Snippet: Description of the SFV RNAs encoding HCV structural proteins and expression in BHK-21 cells. (A) RNAs transcribed from the four SFV DNA constructs. The SFV/Dj construct is our original Dj6.4 clone, encoding HCV core-E1-E2, and is described elsewhere (4). The horizontal line in each construct indicates the segment of the SFV genome. The position of the subgenomic SFV promoter is indicated (26S). Boxes show the inserted genes, and the shaded box in the SFV/Dj-C191 construct indicates the signal sequence targeting the E1 envelope glycoprotein to the ER. SPP, signal peptide peptidase processing between residues 173 and 174. The SFV/Dj-D111A construct was produced by introducing a D111-to-A substitution in the original SFV/Dj construct, i.e., introducing a viral L domain, PTAP. (B) Immunofluorescence staining, using a monoclonal anti-core antibody, of BHK-21 cells electroporated with SFV RNA β-Gal (a), Dj (b), Dj-C191 (c), Dj-C173 (d), and Dj-D111A (e). This assay was performed by following a standard procedure (35), using the mouse monoclonal anti-HCV core antibody MAb 1856 (Virostat, Portland, Maine). (C) Western blotting of these cells with monoclonal anti-core and anti-E2 antibodies. For this standard assay (4), the anti-core MAb 1856 gave poor results. We therefore carried out immunoblotting with the human monoclonal anti-core antibody B12.F8 (11) and the mouse monoclonal anti-E2 (10). Size markers (in kilodaltons) were obtained from New England Biolabs (Beverly, Mass.).

Article Snippet: This assay was performed by following a standard procedure ( 35 ), using the mouse monoclonal anti-HCV core antibody MAb 1856 (Virostat, Portland, Maine). (C) Western blotting of these cells with monoclonal anti-core and anti-E2 antibodies.

Techniques: Expressing, Construct, Sequencing, Produced, Immunofluorescence, Staining, Western Blot