Schematic of dual-function reporter vectors used in the HCV NS3/4A protease activity assay.
- Publication date
- Publisher
Abstract
<p>(A) The various recognition sites of NS3/4A protease, IPS<sub>462–540</sub>, NLS-IPS<sub>462–540</sub>, KDEL-DE<sub>4x</sub>-4A/4B and DE<sub>4x</sub>-4A/4B were inserted between the <i>EGFP</i> and the humanized <i>Gaussia luciferase</i> gene by an in-frame fusion. Arrows indicate the NS3/4A cleavage site. Expression of the reporter genes is under the human cytomegalovirus promoter (CMV) and the selection marker for the generation of the stable cell line is neomycin phosphotrasferase (Neo<sup>R</sup>). SV40, simian virus 40. (B) Transiently trasfected Huh-7.5 cells were infected with JC1 virus at an MOI 0.5 TCID<sub>50</sub>/cell or mock infected with JFH-1/ΔGDD supernatant. At 5 days post infection, the medium was harvested and <i>Gaussia</i> activity was measured. Results are expressed as the mean values from duplicate wells, measured in duplicates, from a representative experiment of 3 (mean ± SD; n = 4).*, <i>P</i><0.05.</p