中间偃麦草转录因子TiERF1a的结合与转录调控特性的研究  

Characteristics of the Transcription Activation and DNA-Binding of An ERF Transcription Factor of Thinopyrum intermedium

作　　者：梁红霞[1] 刘红霞[1] 张增艳[1] 王丽丽[2] 辛志勇[1] 

机构地区：[1]中国农业科学院作物科学研究所/农作物基因资源与基因改良国家重大科学工程/农业部作物遗传育种重点开放实验室,北京100081 [2]河北科技大学生物科学与工程学院,河北石家庄050018

出　　处：《作物学报》2007年第10期1720-1723,共4页Acta Agronomica Sinica

基　　金：国家自然科学基金项目(30671292)

摘　　要：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GCCbox/mGCCbox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GCCbox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 GCC box binding characteristic of an ERF transcription factor TiERF1a isolated from Thinopyrum intermedium,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.

关 键 词：ERF转录因子 转录激活特性 GCC box顺式元件 LACZ基因 Β-半乳糖苷酶 

分 类 号：Q943.2[生物学—植物学]