摘要
利用从烟草中克隆的一个新糖基转移酶(Glucosyitransferase,GTs)基因启动子中具有茉莉酸甲酯(MeJA)和水杨酸(SA)双重诱导的片段,与GUS(β-glucuronidase)报告基因连接,构建部分启动子片段缺失载体。利用农杆菌介导的叶盘转化法,获得了转基因植株,不同片段载体转基因植株的阳性率为50%～71%。对阳性转基因烟草植株GUS诱导活性的定量分析结果表明,在构建的4个表达载体中,启动子片段BA的基础表达水平非常低,并且不同转基因植株中都存在SA和MeJA双重诱导特征;片段BB和BC基础表达水平较高,不同转基因植株对SA和MeJA的反应不同;对于启动子片段BD,基础表达高于BA片段而低于BB和BC,不同转基因植株GUS活性均受SA诱导。推测在-756～-703之间(BA)存在MeJA和SA双重诱导元件,在-643～-606存在SA的诱导元件。对这些诱导元件的确定,有助于水杨酸和茉莉酸甲酯诱导机理的研究。
The glucosyhransferase gene promoter fragment B, which cloned from tobacco by the lab early and induced by both methyl jasmonate and salicylic acid, was fused to the 5＇-upstream of GUS （β-glucuronidase） coding region in binary vector, designated pBA, BB, BC and BD-GUS. The pB-GUS was introduced into W38 tobacco plant by Agrobacterium tumefaciens. The positive ratio of the transgenie tobacco plants with different promoter fragments was 50%-71%. Quantification analysis of GUS activity induced showed that the basic expression of the promoter fragment BA was very low, and the expression was strongly induced by both MeJA and SA. The basic expression of the promoter fragments BB and BC was relatively high, and the different transgenic plants with pBB and BC-GUS showed different response to MeJA and SA. The basic expression of the fragment BD was higher than that of BA, but lower than that of BB and BC, and the GUS activity in transgenic plants with pBD-GUS all were induced by SA. These results showed that in the region of -756--703 （BA） might be there was an element induced by both MeJA and SA, and -643--606 with an element induced by SA. The identification of these elements in this promoter would help to know the principle of inducing mechanism by MeJA and SA.
出处
《湖北农业科学》
北大核心
2009年第9期2058-2060,共3页
Hubei Agricultural Sciences
基金
湖北省自然科学基金项目(2004ABA123)
湖北省高层次人才科研基金项目(QZY03001)
关键词
烟草
糖基转移酶基因
启动子
诱导表达
tobacco
glucosyitransferase gene
promoter
induced expression
