Fig. 3
From: PCR-based reverse genetics strategy for bluetongue virus recovery
IFA was employed to confirm the recovery of infectious BTV-1 (strain SZ97/1) by transfection of BSR cells with 10 PCR amplicons and helper plasmids. The lysates from the transfected cell, mock-transfected cell, helper plasmids only transfection cell and PCR products only transfection cell were harvested and infected in BHK-21 cells; 4 days post infection, IFA was performed with anti-BTV VP7 monoclonal antibody (named BTV-4H7) and goat anti-mouse IgG FITC labelled antibody. If BTV were recovered successfully, the BHK-21 cell was further infected, and the green florescence signal could be detected with IFA. a Green fluorescence signal was detected in BHK-21 cells which were infected with experimental group. b No fluorescence was observed in BHK-21 cells infected with mock-transfected cells lysate. c No fluorescence was observed in BHK-21 cells infected with helper plasmids only transfected cells lysate. d No fluorescence was observed in BHK-21 cells infected with PCR products only transfected cells lysate