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Fig. 3 | Journal of Biomedical Science

Fig. 3

From: Naturally occurring mutations in PB1 affect influenza A virus replication fidelity, virulence, and adaptability

Fig. 3

Effects of the PB1-S216G mutation on replication capability and mutation potential in virus-infected cells using the dual-luciferase RT2AF reporter. a Schematic diagram of mutability assay for influenza RdRp. The dual-luciferase RT2AF reporter is flanked by the 5′ and 3′ UTR sequences of the WSN-NP genome, and transcription was controlled by the human PolI promoter and the murine terminator. b Replication capability was calculated based on Rluc luciferase activity and c the mutation potential was calculated as cumulative mutation index (CMI) based on the Fluc/Rluc ratio. d and e Mutation potential of RdRp from PB1–216 variants was measured by influenza minireplicon system. The PB2, PA, NP expression plasmids plus wild-type PB1 or PB1–216 variant plasmids were co-transfected with RT2AF reporter in HEK 293 cells. After 72 h, the replication capability (d) by Rlu luciferase activity and mutation potential (Cumulative Mutation Index; CMI) (e) by Fluc/Rluc ratio were evaluated in the indicated PB1 plasmids containing either 216S or 216G, respectively. Error bars indicate the standard error of the mean of three independent experiments. Student’s two-tailed unpaired t-test was performed to determine the P value; NS, not significant (p > 0.05)

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