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为了进一步了解VER2基因的表达模式及其启动子的功能,从小麦可转化的人工染色体基因组文库中获得41.7 kb含VER2基因的TAC克隆,序列分析显示,其含有11个推测基因,其中第3个基因的外显子完全与VER2基因的cDNA序列同源.VER2基因的启动子存在3个小的重复序列,被另外2个大重复序列所分割.对上游启动子区的响应元件分析结果表明,其包含ABA响应元件(ABRE)、茉莉酸甲酯响应元件(Me-JARE)、低温响应元件(LTR)、胚乳特异性表达元件以及参与淀粉酶合成的元件和类似GA响应元件等.利用基因枪的方法,将VER2启动子(-5895-+73)驱动GFP报告基因的瞬间表达载体转入经春化处理或未春化处理的小麦幼叶中,结果发现,GFP在春化处理的幼叶中表达,而在未春化处理的幼叶中不表达,说明春化作用是VER2启动子在冬小麦幼叶中驱动基因转录所必需的.
In order to further understand the expression pattern of VER2 gene and the function of its promoter, a 41.7 kb TAC clone containing the VER2 gene was obtained from the genome of the wheat genome that can be transformed. Sequence analysis showed that it contained 11 putative genes, of which 3 The exon of the gene is completely homologous to the cDNA sequence of VER2 gene.The three small repeat sequences of the promoter of theververg 2 gene are segregated by two other large repetitive sequences.The analysis of the response elements to the upstream promoter region shows that, It contains ABRE, Me-JARE, LTR, endosperm-specific expression elements, amylase-involved elements and similar GA-responsive elements, etc. Using gene gun The transient expression vector of GFP reporter gene driven by VER2 promoter (-5895- + 73) was transferred into young leaves of vernalized or vernalized wheat leaves. , But not in the vernalized young leaves, indicating that vernalization is necessary for the VER2 promoter to drive gene transcription in young leaves of winter wheat.