Figure 2

Biochemical function of OsNAP in yeast. A, Constructs for the yeast one-hybrid assay. B, OsNAP can bind to the NACRS-like sequence in yeast. The pGAD-OsNAP plasmid and reporter construct pHIS-cis[34] were cotransformed into the Y187 yeast strain. The transformants were examined by growth performance on SD/Leu-/Trp-/His- plates with or without 30 mM 3-AT. N, negative control (p53HIS2 plus pGAD-OsNAP); P, positive control (p53HIS plus pGAD-Rec2-53); lables 1–4, four different colonies containing pGAD-OsNAP and pHIS-cis. C, Constructs of transactivational analysis. D, Transactivation assay of OsNAP in yeast. The full-length and partial fragments of OsNAP were ligated into the pGBDKT7 vector and co-transformed into the AH109 yeast strain with pGADT7 to verify the transactivation activity. The transformants were examined by growth performance on SD plates in the absence or presence of histidine and the necessary amino acids. N, negative control (pGBDK7/pGADT7); P, positive control (pHIS53/pGAD-Rec2-53); lables 1–3, three different colonies containing positive control, pGBD-OsNAP/pGADT7, pGBD-OsNAP-ABC/pGADT7, pGBD-OsNAP-DE/pGADT7, pGBD-OsNAP-TR/pGADT7 and negative control, respectively.