Yeast two-hybrid analyses–P. brasiliensis/P. brasiliensis gene interactions.

Colonies that grew on SD-Ade/-His/-Leu/-Trp drop-out plates and had α-galactosidase activity, as determined by a blue colouration of the colonies growing on α-X-gal supplemented media, are defined as positive (+) and those that did not as (-). All the gene sequences were swapped between the pGADT7 and pGBKT7 vectors and only those giving a positive-reaction in both vectors were scored as positive for an interaction. In addition, Tpk2(1–583) was tested for interactions with PbCyr1(600–1316), PbCyr1(1302–1871), PbCyr1(1649–2100) but none were detected. A set of control reactions was undertaken at the same time to validate the test reactions: positive control, pGADT7- Ag with pGBKT7-P53; negative controls, pGBKT7-Lam with pGADT7, pGADT7-TPK21-270, pGADT7-TPK21-583, pGADT7-TPK2265-583 and pGADT7-GPG1, and pGBKT7-TPK1 with pGADT7, pGBKT7, pGADT7-Lam, pGBKT7-Lam and pGBKT7-PbActin. NT- Not tested.Yeast two-hybrid analyses–P. brasiliensis/P. brasiliensis gene interactions.