Figure 4

Expression of His-tagged GFP (27 kDa) in the leaves of N. benthamiana with NMV viral vector pNMV-GFP and pNMV-GFPer. A: Image taken under UV light, showing systemic distribution of GFP without basic chitinase signal peptide at the N-terminus and the ER-retention peptide HDEL at the C-terminus; B: Image taken under UV light, showing systemic distribution of GFP containing basic chitinase signal peptide at the N-terminus and the ER-retention peptide HDEL at the C-terminus; C: total proteins were extracted from the leaves inoculated with pNMV-GFP and pNMV-GFPer respectively and separated on a 4-20% SDS-PAGE gel, transferred onto PVDF membrane and probed with Anti-GFP rabbit IgG. Lane 1: Protein Marker; Lane 2: Mock control; Lane 3: leaves inoculated with pNMV-PAP1, Lane 4: inoculated with pNMV-hGFP; Lane 5: the leaves inoculated with pNMV-hGFPer; Lane 6–10 were the same samples as in Lane 1–5 but having gone through Ni-Sepharose affinity purification. D: Identical Western blot as in C but probed with mouse monoclonal His-Tag antibody as the primary antibody.