一个小麦AP2／ERF转录因子家族单独亚族基因的克隆及分析

Cloning and Analysis of an AP2/ERF Soloist Subfamily Transcription Factor from Triticum aestivum L.Yangmai 15

AP2/ERF家族转录因子是植物中重要的一类转录因子,广泛参与植物各类生理过程。为了给该家族转录因子的深入研究提供基础,利用小麦EST数据库(http://www.ncbi.nlm.nih.gov/UniGene/UGOrg.cgi?TAXID=4565),以拟南芥AP2/ERF家族转录因子单独亚族成员AtAP2-Soloist的氨基酸序列为探针,搜索到一个小麦UniGene数据,经拼接得到1个全长cDNA序列(TaAP2-Solosit),通过RT-PCR方法从"扬麦15"的cDNA中克隆了该转录因子基因(YM15 AP2-Solosit)。克隆的转录因子核苷酸序列与电子克隆的序列只有1个碱基不同,两者编码氨基酸序列一致。从cDNA序列、氨基酸序列的相似性、组成成分、理化性质、疏水性/亲水性、进化树、功能域、二级和三级结构等方面进行了分析,结果显示"扬麦15"中的YM15 AP2-Solosit转录因子属于AP2/ERF家族中的单独亚族,是亲水性蛋白,在蛋白质的三级结构上与拟南芥的AtAP2-Soloist相似。EST丰度分析显示,YM15 AP2-Solosit在根、茎、叶和花中均有表达。

AP2/ERF is a large family of plant transcription factors. Genes in the AP2/ERF family encode transcriptional regulators that serve a variety of functions in the plants. An AP2/ERF family transcription factors （TaAP2-Solosit） was isolated by silico cloning method using the AtAP2-Soloist amino acid sequence of Arabidopsis thaliana as probe from wheat （Triticum aestivum L. ） ESTs （http：//www. ncbi. nlm. nih. gov/UniGene/UGOrg, cgi？ TAXID = 4565）. Based on the sequences of TaAP2-Solosit, YM15AP2-Solosit gene was isolated from wheat Yangmai 15 by RT-PCR using cDNA as template. Only one oligonucleotide was different between TaAP2-Solosi and YM15AP2-Solosit but their amino acid sequences were the same. Then, cDNA and deduced amino acid sequence, composition, hydrophobicty and hydrophilicity, physical and chemical characterization, phylogenetic tree, conserved domain sequences,function domain and molecular modeling were analyzed. YM15AP2-Solosit was belonged to Solosit sub family of AP2/ERF family. YM15AP2-Solosit was hydrophilic protein. TaAP2-Solosi and YM15AP2-Solosit have similar three-dimension structure. YM15AP2-Solosit was expressed in seed, stem, leaf and in florescence.