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. 2014 Jan 14;450:290–296. doi: 10.1016/j.virol.2013.12.018

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Fig. 2 — Effects of marafibox and major coat protein (CP) gene mutations on accumulation of viral CPs, and genomic and sgRNAs. (A) Nucleotide sequences from known (OBDV, MRFV, BELV, CSDaV) and proposed (SwMV, BlVS, GAMaV, GRVFV, GSyV-1) marafiviruses aligned with those of PnMV (unassigned) and TYMV (type member of the genus Tymovirus). Silent substitutions (U>C, G>A) introduced to produce mutant OBDV-AB15-25 are shown and the adenine position representing the proposed start site of sgRNA synthesis is indicated (bent arrow); underlined adenine residues below the horizontal arrow have been experimentally confirmed as sgRNA 5'-ends. (B) Accumulation of marafibox and major CP mutants in oat protoplasts. Oat protoplasts were inoculated with capped transcripts of wild type clone pOBDV, major CP mutants GH1-7 (premature termination mutant) and KL1-3 (initiation codon mutant), and AB15-25 (marafibox mutant) and incubated for 40 h. Extracted viral RNA and protein were detected on northern and western blots using a 3'-end dsDNA probe and anti-OBDV antiserum, respectively. The locations of major CP (Maj. CP), minor CP (Min. CP), genomic (gRNA), and subgenomic RNA (sgRNA), on the blots are shown.