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. 2026 Apr 21;100(5):e00119-26. doi: 10.1128/jvi.00119-26

Fig 6.

Dual-luciferase assays and Western blots identifying RNA elements crucial for PRRSV -1 ribosomal frameshift. Results demonstrate 75 nucleotide minimal sequence requirement and essential pseudoknot structure for efficient frameshifting.

Mapping the RNA signal required for programmed −1 ribosomal frameshifting in PRRSV. (A) Schematic diagram of the dual-luciferase reporter carrying the putative −1 PRF signal or its mutants. (B and C) The minimal sequence required for efficient PRRSV –1 PRF is 75 nt. Truncated mutants of –1 PRF or IFC were transfected into HEK-293T cells. At 24 hpt, cell lysates were harvested for the dual-luciferase assay (B) and Western blot analysis (C). (D and E) Mutational analysis of the predicted pseudoknot structure. The WT plasmid, pseudoknot mutants, or IFC were transfected into HEK-293T cells. At 24 hpt, cell lysates were analyzed by dual-luciferase assay (D) and Western blot analysis (E). The expression of the pDluc translation products was detected by an antibody recognizing Renilla luciferase. β-Tubulin was detected as a loading control. Data are presented as means ± SD from three replicates. Statistical significance was analyzed using the two-tailed unpaired Student’s t-test. ****, P < 0.0001; ns, not significant.