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Fig. 4 | Virology Journal

Fig. 4

From: Capped antigenomic RNA transcript facilitates rescue of a plant rhabdovirus

Fig. 4

Function of N proteins translated in cis from capped agRNA in SYNV rescue. a Diagram showing insertion of the GFP gene between the leader and N genes (SYNV-GFPle/N) or between the N and P genes (SYNV-GFPN/P) in SYNV agRNAs. GFP mRNA transcription was directed by a duplicated leader/N or N/P gene junction sequences, in SYNV-GFPle/N or SYNV-GFPN/P, respectively. b and c Transient expression of genes located at the 5′ termini of SYNV agRNAs. Total protein samples were exacted from N. benthamiana leaves at 5 dpi after agroinfiltration with 35S-HH3-SYNV and 35St-SYNV (b), or 35St-SYNV-GFPle/N and 35St-SYNV-GFPN/P (c). Agrobacteria carrying VSR plasmids were also included in the infiltration mixtures to suppress RNA silencing. Protein gels were blotted with anti-N and anti-GFP antibodies. d Symptoms of recombinant SYNV-GFPle/N- and SYNV-GFPN/P-infected plants. Recombinant viruses recovered from N. benthamiana plants were passaged to healthy plants by sap inoculation. The infected plants were photographed at 16 dpi under visible light (upper panels) and ultraviolet (UV) light (bottom panels). e Immunoblot analysis of SYNV structural proteins and GFP expression in systemically infected tissues of plants inoculated with recombinant SYNV-GFPle/N and SYNV-GFPN/P using antibodies against disrupted SYNV virions or GFP. In (b), (c) and (e), the protein blots were stained with Ponceau S to verify similar level of the large subunit of Rubisco (Rub L)

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