FIG 8.

Analysis of viral RNA synthesis of noncp and cp BVDV NS2-5B RNA replicons in MDBK DNAJC14-KO cells. (A) Scheme of the bicistronic reporter replicons Bici RLuc NS2-5B/CP7 (top) and Bici RLuc NS2-5B/NCP7 (bottom). The 9-aa insertion in NS2 of Bici RLuc NS2-5B/CP7 is indicated as a light gray box within the NS2 gene. S (black box) depicts the C-terminal portion of p7 (38 aa) that acts as a signal sequence for translocation of NS2-3 (10). (B) Cells were electroporated with 1 μg of the corresponding in vitro-transcribed RNAs and incubated for the indicated times at 2, 24, and 48 hpe, and luciferase activity was determined. Measurements were carried out in triplicate, and experiments were repeated at least 3 times. Data from one representative experiment are shown. Error bars represent standard deviations. Mock, no RNA electroporated; GAA, NS5B mutation, replication deficient; WT, wild type; RLUs, relative light units; RLuc, Renilla luciferase.