Knock-Down of Both eIF4E1 and eIF4E2 Genes Confers Broad-Spectrum Resistance against Potyviruses in Tomato
Figure 6
Interaction assays between the three eIF4E proteins from tomato and a selected set of VPg from potyviruses using the yeast two-hybrid system.
Yeast expressing both “bait” and “prey” recombinant proteins were obtained by first transforming yeast cells with the individual plasmid construction followed by separate conjugation between yeasts expressing eIF4E proteins and those expressing VPg proteins. Conjugations were then cultured on control plates (SD-LW) and on two selective media lacking leucine, tryptophan and histidine (SD-LWH) or leucine, tryptophan, histidine and adenine (SD-LWHA). Each plasmid combination was spotted in duplicate. Positive and negative controls from the Matchmaker GAL4 two-hybrid system 3 were used (bottom panel).