摘要
组织特异性启动子是植物基因工程改良的重要工具。RAFTIN是禾本科植物花药绒毡层中特异积累与花药发育相关的蛋白,其调控序列可能提供一个很好来源的花药特异性表达启动子。为了解小麦TaRAFTIN1a基因启动子的花药表达特异性,本研究分离了TaRAFTIN1a基因的启动子,生物信息学分析表明,该启动子含有3种花药特异表达元件(总计10个),1个ABRE脱落酸响应元件,TGACG-motif与CGTCA-motif茉莉酸甲酯响应元件各1个,8个ARR1AT细胞分裂素响应元件,以及2个DRE和1个MBS干旱响应元件。采用5'端缺失的方法,分别克隆1429、898和351 bp的启动子片段,经连接、转化与鉴定,构建了3个含TaRAFTIN1a启动子不同区段融合GUS报告基因的植物表达载体,p WAER1,p WAER2,p WAER3,为通过转基因了解启动子的表达特征及其核心作用元件奠定了基础。
Tissue-specific promoters are important tools in plant genetic engineering. The occurrence of RAFTIN protein is limited to cereals,which is located only in the tapetum and involved in the anther development. Its regulation related sequence could be a good source for plant anther-specific promoter. In order to understand the anther-specific expression of wheat homolog TaRATIN1 a,the promoter of TaRAFTIN1 a was isolated in this study and analyzed with the method of bioinformatics analysis. The result showed that three types of anther-specific expression elements（ 10 in total） were found out,including one ABA-response element ABRE,8 CK-response element ARR1 AT and two kinds of drought respond element,DRE and MBS,respectively. Through 5＇end deletion,TaRATIN1 a promoter with a variation of length,1429,898 and 351 bp,were ligated with GUS reporter gene. After identification and transformation of recombinant,three plant expression vectors,named as p WAER1,p WAER2 and p WAER3,respectively,were constructed,which would be used for the investigation on expression character and the key regulatory elements of the promoter by transgenic experiments.
出处
《西南农业学报》
CSCD
北大核心
2015年第1期1-5,共5页
Southwest China Journal of Agricultural Sciences
基金
中国科学院院地合作项目(XBCD-2011-019)
中国科学院知识创新工程重要方向项目(KSCX2-EW-J-22)
四川省科技厅国际合作项目(2012HH0008)
关键词
普通小麦
花药特异表达启动子
启动子分析
载体构建
Triticum aestivum
Tissue-specific promoter
Promoter analysis
Plant expression vector
