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植物研究  2015 

小黑杨PnsICE1基因启动子克隆及瞬时表达分析

DOI: 10.7525/j.issn.1673-5102.2015.01.013, PP. 77-83

Keywords: 小黑杨,PnsICE1启动子,顺式调控元件,逆境胁迫

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Abstract:

?ICE1基因是能够编码类似MYC的bHLH转录因子,在低温条件下能够特定地结合CBF3启动子中的MYC作用元件,继而诱导CBF3调控的下游基因的表达,在植物逆境胁迫应答中具有重要的功能。本研究通过PCR扩增技术从小黑杨(Populussimonii×P.nigra)基因组DNA中克隆得到1247bpICE1基因启动子序列,在线预测分析发现,该段序列中含有典型的真核生物核心启动子区域,除了包含多个TATA-box、CAAT-box等基本顺式作用元件外,还存在其它反应元件,如ABRE、DOFCOREZM、MYBCORE、W-box和MYCCONSENSUSATHSE等,这些元件有的与赤霉素、脱落酸等激素相关,有些与逆境胁迫诱导相关,这表明PnsICE1基因启动子可能与逆境胁迫相关,在小黑杨抵御逆境胁迫的生理过程中具有重要功能。将克隆得到的PnsICE1启动子取代pBI121中的CaMV35S启动子,构建植物表达载体pBI121-ICEPro,由PnsICE1启动子驱动报告基因gus表达,并用农杆菌介导的瞬时侵染法转化拟南芥。对瞬时侵染后的拟南芥进行GUS染色分析。结果表明,在PnsICE1启动子的驱动下,报告基因gus在花及根部中特异表达,PnsICE1启动子具有启动活性。

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