The gene portion containing prM-E, as described in Puttikhunt, et al [24] , was amplified from DENV2 (NGC) cDNA using the primers D2-FW (59-AATTAATACGACCGTCTCCCATGAA-TAGAAGACGCAGATCTGCAGGC-39) and D2-RV (59-CGC-CCGTTTGATCTCGAGCTACTAGGCCTGCACCATGACT-CCC-39) and cloned into pCMV-myc-ER vector via the restriction sites Nco I and Xho I to create plasmid construct pCMV-prM-E. Using this as template, selected amino acids residues of the predicted epitopes, P3 and P9, were mutated by site-directed mutagenesis using Quikchange Multi Site-directed Mutagenesis kit (Stratagene, Agilent Technologies). PCR reactions and subsequent cloning steps were carried out according to manufacturer's instructions. Substitution of amino acids in all mutant constructs was confirmed by sequencing. Mutants were expressed in HEK293-T cells and harvested 48 hr posttransfection for immunoblot analysis and immunoprecipitation experiments.