Abstract:

In order to investigate the transactivation activity and interaction characteristic of an ERF transcription factor TiERF1a isolated from Thinopyrum intermedium and its cis-acting element GCC box, we successfully constructed the yeast reporter vectors containing 3 tandem copies of GCC box or mutated GCC box fused upstream of lacZ gene, which had been integrated into the genome of the yeast strain YM4271 and the effecter expression vector pYepGAP-TiERF1a containing the full encoding region of TiERF1a gene. The TiERF1a expression vector pYepGAP-TiERF1a was transformed into the reporter yeast cells. The activity of lacZ gene expression product,β-galactosidase, was assayed qualitatively by X-gal colony-lifted filter, and analyzed quantitatively using o-nitrophenyl-β-D-galactopyranoside (ONPG) as substrate. The results indicated that TiERF1a could bind to GCC box sequence and possessed high transactivation activity. The method established can also be used to analyze the transcriptional regulation activity of other transcription factors in yeast cells.

Key words: ERF transcription factor, transactivation, GCC box cis-acting element, lacZ gene, β-galactosidase