FIG 8.
Validation of interactions between GN and TSWV-interacting proteins (TIPs) and identification of the interacting domain of endoCP-GN using a split-ubiquitin membrane-based yeast two hybrid (MbY2H) assay. (A) Interactions between GN and six TIPs. GN was expressed as GN-Cub, and TIPs were expressed as NubG-TIPs using MbY2H vectors. (B) Interactions between TSWV GN and different regions of endoCP-GN. EndoCP-GN was expressed as either the N-terminal domain (amino acids 1 to 176 and 1 to 189) that includes the nonconserved region or the C-terminal region (amino acids 177 to 284 and 190 to 284) that includes the conserved Chitin_bind_4 motif (CHB4) of endoCP-GN. Interactions between GN-Cub and NubI and between GN-Cub and NubG were used as positive and negative controls, respectively, for all MbY2H assays. Cotransformation of pTSU2-APP and pNubG-Fe65 into NYM51 was used as another positive control (data not shown). DDO, yeast double-dropout (SD/−Leu/−Trp) medium; QDO, yeast quadruple-dropout (SD/−Ade/−His/−Leu/−Trp) medium.