构建了含中间偃麦草ERF转录因子基因TiERF1a的酵母表达载体pYepGAP-TiERF1a及含GCC box/ mGCC box和lacZ基因的酵母报告子，将pYepGAP-TiERF1a成功转到报告子酵母细胞中，对转化的酵母细胞内lacZ基因表达产物(β-半乳糖苷酶)活性进行定性和定量分析。结果表明，TiERF1a在酵母体内能与GCC box顺式元件结合并激活下游lacZ基因的表达。本方法还可用于比较和研究其他转录因子的转录调控特性。
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.
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