Fig. 6: Analysis of AaCBF4 sequence and protein characterization.

a AaCBF expression was analyzed under cold stress. b Transactivation assay of CBFs in yeast. The fusion proteins of the GAL4 DNA-binding domain (BD) and full-length CBFs were expressed in yeast strain AH109. The empty vector pGBKT7 was used as a negative control. A culture solution of the transformed yeast was plated on SD/-Trp solid media, SD/-Trp/-Ade/-His solid media and SD/-Trp/-Ade/-His solid X-α-Gal solid media, as indicated. c. Subcellular localization of AaCBF4 in tobacco. Plant leaves were injected with Agrobacterium tumefaciens GV3101 containing the AaCBF4:eGFP vector. After 3 days, the leaves were observed for fluorescence with a confocal microscope (FV1000; Olympus, Tokyo, Japan). The nuclei were stained with DAPI