Fig. 3

Analysis of transcriptional activity of ZmCCT protein. a) Transactivation analysis of ZmCCT fused to the GAL4 DNA-binding domain in yeast. The fusion vectors pGBKT7-ZmCCT, pGBKT7-Gal4 (GAL4) and pGBKT7 (BD) were respectively transformed into the yeast competent cells. Firstly, the deficient medium SD/−Trp was screened, and then the positive monoclones were selected for verification in the deficient medium SD/−Trp/−His/X-gal and SD/−Trp/−His/−Ade. GAL4 was the positive control and BD was the negative control. b) Sequence alignment of CCT-domain from ZmCCT, OsGHD7 and eight Arabidopsis CCT-domain containing gene. The alignment was performed using DNAMAN. c) Determination of ZmCCT transcriptional activity. The reporter (upper) and effector constructs (left) were co-transformed into N. benthamiana leaves, and the expression level of the reporter gene was determined (right) (n = 5, ±SD, ** P < 0.01). VP16: VP16-LexA fusion protein; Gal4: Gal4 DNA binding domain (DBD); CCT: full-length ZmCCT fused to Gal4 DBD