蒙古沙冬青AmDREB2.2基因的克隆与植物表达载体构建  

Cloning of AmDREB2.2 Gene of Ammopiptanthus mongolicus and Construction of Its Plant Expression Vector

作　　者：张至玮 李章磊[1] 高飞[1] 曹玉震 夏波林 周宜君[1] 

机构地区：[1]中央民族大学生命与环境科学学院,北京100081

出　　处：《湖北农业科学》2015年第16期4065-4069,共5页Hubei Agricultural Sciences

基　　金：国家自然科学基金项目(31370356);中央民族大学"985工程"学科建设项目(YLDX01013);国家大学生创新训练项目(GCCX2014110020);中央民族大学研究生科研创新项目(K2014044)

摘　　要：从前期建立的蒙古沙冬青[Ammopiptanthus mongolicus(Masxim)Cheng f.]根转录本数据库中克隆得到1个编码DREB类转录因子基因。结果表明,Am DREB2.2基因序列全长1 045 bp,开放阅读框(ORF)为597 bp,编码198个氨基酸,具有典型的DREB转录因子保守的AP2结构域。实时荧光定量PCR分析表明,该基因能在根、叶中表达,但对干旱、低温响应不同,Am DREB2.2主要参与根的干旱胁迫应答。在Am DREB2.2基因的编码区两端分别加入Bam HⅠ和SacⅠ酶切位点,双酶切植物表达载体p PZP212和带有酶切位点的Am DREB2.2基因PCR产物,成功获得了Am DREB2.2的植物过表达载体p PZP212-Am DREB2.2。A new dehydration responsive element binding protein（DREB） transcription factor gene, which was named as AmDREB2.2, was cloned from Ammopiptanthus mongolicus（Masxim.）（cheng f.） root transcriptome database. The full length of the Am DREB2.2 c DNA was 1 045 bp, including a single 597 bp opening reading frame which encoded a 198-amino acid peptide with a conserved AP2 domain. Quantitative real-time PCR analysis revealed that the Am DREB2.2 expressed in leaf and root of A. mongolicus,but there were different expression patterns under drought or low temperature stress respectively,and it could be mainly induced by drought in root. The plant expression vector p PZP212 and Am DREB2.2 with Bam H I and Sac I site were digested by these two restriction enzymes, were ligated and recombinant, and then plant expression vector p PZP212-Am DREB2.2 was successfully constructed.

关 键 词：蒙古沙冬青[Ammopiptanthus mongolicus(Masxim)Cheng f.] AmDREB2.2基因 表达特征 植物表达载体 

分 类 号：Q78[生物学—分子生物学]