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. 2010 Jun 16;84(17):8849–8860. doi: 10.1128/JVI.00456-10

FIG. 3.

FIG. 3.

miR-323, miR-491, and miR-654 bind to the same region in the PB1 gene. (a) The sequences of miR-323, miR-491, miR-654, and PB1-5 mRNA. The same nucleotide sequence in miRNAs 5′-GUGG-3′ perfectly matches the 5′-CCAC-3′ sequence in the PB1 mRNA. (b) The model of hybridization between the miRNAs and PB1-5 mRNA were predicted using RNAHybrid software. (c) The mutant nucleotide sequence of pRL-TK-PB1-5 and sequencing analysis. (d) The mutant nucleotide sequences of the miRNAs. The mutant 5′-UUCC-3′ sequences in ΔmiRNAs perfectly match the 5′-GGAA-3′ sequence in ΔpRL-TK-PB1-5 mRNA. (e) Luciferase assay to analyze the importance of binding sites of miRNAs in the PB1 gene for miRNAs to inhibit the relative luciferase activity from pRL-TK-PB1-5 and pGL3-control vector.