FIG 2.

Characterization of Rluc-ZIKV-T154C reporter virus uncoupling the DAR element from the capsid-coding region. (A) Schematic representation of the Rluc-ZIKV-T154C construct. The N-terminal 38 amino acids of the capsid (C38) were duplicated downstream of the 5ʹ UTR, and the Rluc gene fused with FMDV 2A sequence (Rluc-2A) was inserted at the C38 and capsid junction. In the full-length capsid, a nucleotide mutation, T154C, within the 5ʹ CS sequence was introduced to limit the interaction between the 5ʹ and 3ʹ CS. (B) IFA analysis of ZIKV-T154C and Rluc-ZIKV-T154C. Equal amounts of the in vitro-transcribed RNAs of Rluc-ZIKV-T154C, ZIKV-T154C, and ZIKV-WT were transfected into Vero cells, and IFA analysis was performed at the indicated time points posttransfection. The polyclonal antibody against NS3 and FITC-conjugated goat anti-mouse IgG were used as primary and secondary antibodies, respectively. (C) Production of ZIKV-WT, ZIKV-T154C, and Rluc-ZIKV-T154C at 24, 48, and 72 hpt. The data are representative of three independent transfections, and the error bars represent standard deviations.