Figure 8. Yeast two-hybrid assays screening StMKK6 interaction partners.

StMKK6 protein was fused with Gal DNA-binding domain as bait and StMAPK4_1, StMAPK4_2, StMAPK6 and StMAPK13 were used as prey fused with Gal DNA-activation domain. Interaction pairs P53/SV40 large T-antigen and Lam/SV40 large T-antigen were used as positive and negative controls respectively. StMKK6/empty vector pair was used as a control to discard auto-activation of bait protein. Serial dilutions of interaction pairs were plated on DDO media for co-transformation selection. Interactions of bait and prey proteins were examined by assessing growth on several selective media with different levels of restrictiveness i.e. SD/-Leu/-Trp/-His (TDO), SD/-Leu/-Trp/x-a-Gal/Aba (DDO/X/A) and QDO/X/A. Only co-transformed colonies growing on the most restrictive media (QDO/X/A) were considered as positive interaction transformants.