Fig. 2

Obtaining recombinants plasmids and recombinant NS3 protein. a Cloning strategy for the NS3 domain protease of dengue virus in the cloning vector and in prokaryotic and eukaryotic expression vector. b Purification of the recombinant protein NS3DEN3. SDS-PAGE 10% coomassie blue stained. (M) molecular weight marker. (1) Total cell extract of E. coli DH5α uninduced. (2) Total cell extract of E. coli DH5α induced with IPTG. (3) recombinant NS3DEN3 protein. The arrow points to the purified protein NS3DEN3 (c) Detection of the recombinant NS3 protein by western blot with an anti-NS3 antibody. d (M) size marker. (Lane 1) pcDNA3 plasmid linearized with EcoRI. (Lane 2) recombinant plasmid pcDNA3/NS3DEN3 linearized with EcoRI (Lane 3) Fragment release NS3 pro-185. The arrow indicates the fragment released. e HeLa cells transfected. SDS-PAGE 10% coomassie blue stained. (M) molecular weight marker. (1) Total extract untransfected HeLa cells. (2) total extract of HeLa cells transfected with plasmid pcDNA3/NS3/DEN3. f Detection of NS3 protein by western blot with an anti-NS3 antibody