Fig. 6.

Domain–domain interaction by yeast two-hybrid assay. The bait represents the vector containing the GAL4 DNA binding domain, and the prey represents the vector containing the GAL4 activation domain. a The protein–protein interaction of NtD and NtD could be identified in the pGBT9-NtD and pACT-NtD constructs, whereas the constructs of pGBT9-NtD/pACT and pGBT9/pACT-NtD confirmed that the interaction of NtD and NtD was not caused by autoactivation. Lack of yeast grown with the construction of pGBT9-CtE and pACT-CtE showed that no interactions could be found between CtE and CtE. However, a few yeast colonies in pGBT9-NtD and pACT-CtE showed a weak interaction between NtD and CtE. b The constructs of NtD, CtE and FL were subcloned into pGBT9 and pACT vectors. An interaction could be observed in FL and FL, and NtD and FL. The constructs of pGBT9-FL/pACT and pGBT9/pACT-FL also confirmed that the interaction of FL was not caused by autoactivation. pGBKT7-p53/pGADT7-T and pGBT9/pACT served as the positive and negative control, respectively. Yeast was incubated in the SD/-Trp/-Leu/-His selection medium containing 5 mM 3-AT