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脱落酸(ABA)在植物的生长发育中起着重要的作用,ABI5是响应ABA信号的关键基因,研究调控ABI5基因表达的转录因子对进一步阐述ABA信号转导及调控具有重要意义。本研究中,将3-AF1、Box I和Sp1三次重复串联后连接到pAbAi诱饵质粒上,转化酵母细胞构建诱饵载体,构建桃(Prunus persica)酵母单杂交cDNA文库,再共转化诱饵菌株,经同源重组筛选PpABI5启动子的上游转录调节因子。构建的cDNA文库库容为1×10~7 CFU,插入片段长度平均在1 500 bp左右。酵母单杂交筛选结果经测序和Blast同源性分析,获得PpDAM3和PpDAM5两个转录因子。酵母单杂交进一步证实PpDAM3和PpDAM5能与PpABI5启动子结合。这些研究结果表明,PpDAM3和PpDAM5可能参与调控PpABI5的转录,并为深入研究ABA信号转导通路奠定基础。
Abscisic acid (ABA) plays an important role in plant growth and development. ABI5 is a key gene in response to ABA signaling. Studying transcription factors that regulate ABI5 gene expression is of great significance to further elucidate ABA signal transduction and regulation. In this study, 3-AF1, Box I and Sp1 were inserted into pAbAi bait plasmids in series after being repeated three times in series. The bait vector was transformed into yeast cells to construct a yeast single hybrid cDNA library of Prunus persica. Homologous recombination was used to screen upstream transcriptional regulators of the PpABI5 promoter. The constructed cDNA library has a capacity of 1 × 10 ~ 7 CFU, and the length of inserted fragment is about 1 500 bp. Yeast single hybrid screening results were sequenced and Blast homology analysis, access to PpDAM3 and PpDAM5 two transcription factors. The yeast single hybridization further confirmed that PpDAM3 and PpDAM5 can bind to the PpABI5 promoter. These findings suggest that PpDAM3 and PpDAM5 may be involved in the regulation of PpABI5 transcription and lay the foundation for further study of ABA signal transduction pathway.