A Novel System for Rapid and Cost-Effective Production of Detection and Diagnostic Reagents of West Nile Virus in Plants
Figure 1
Western blot analysis of WNV DIII antigen and E16 mAb expression in N. benthamiana and lettuce. (a) WNV DIII. Leaf protein extracts were separated on a 4–20% SDS-PAGE gradient gel and transferred onto PVDF membranes. The membranes were probed with a rabbit anti-WNV DIII polyclonal antibody. Lane 1: E. coli-derived DIII standard; lane 2: protein extract from leaves infiltrated with buffer (negative control); lane 3: extract from DIII construct infiltrated N. benthamiana leaves. (b) and (c) Expression of E16. Wild-type laboratory-grown lettuce or N. benthamiana were infiltrated with dual-replicon geminiviral vector pBY-HL (hE16-no-KDEL). R and harvested on 4 dpi. Total leaf protein extracts were separated on 4–20% SDS-PAGE gradient gels under reducing conditions and transferred to PVDF membranes. The membranes were incubated with a goat anti-human-gamma chain antibody to detect HC (b) or a goat anti-human-kappa chain antibody to detect LC (c). Lane 1: human IgG reference standard; lane 2: extract from lettuce leaves infiltrated with buffer (lettuce negative control); lane 3: protein samples from lettuce infiltrated with geminiviral vector pBY-HL(hE16-no-KDEL). R; lane 4: extract from buffer-infiltrated N. benthamiana leaves (negative control); Lane 5: N. benthamiana leaf protein extract infiltrated with pBY-HL(hE16-no-KDEL). R vector.
