马蔺重金属ATP酶基因HMA 2的RNAi载体构建及其遗传转化

发布时间：2018-06-17 02:10

本文选题：马蔺 + 重金属ATPase酶　；参考：《草业科学》2017年05期

【摘要】：重金属ATP酶基因HMA2介导的Cd长距离运输对植物体内Cd的分配及解毒起重要作用。然而,有关马蔺(Iris lactea)体内Cd长距离运输的分子机制仍不清楚。本研究以马蔺根中总RNA为模板,采用RT-PCR方法获得了重金属ATP酶基因片段,以中间载体pHANNIBAL和植物表达载体pART27为基础,采用酶切和连接的方法,构建了CaMV35S启动子驱动的含有IlHMA2基因片段反向重复序列的RNAi植物表达载体pARTG1G2,用冻融法将此重组质粒导入农杆菌GV3101,并转入到马蔺幼苗体内获得了IlHMA2-RNAi转化株系。这将为阐明IlHMA2介导的Cd长距离运输在马蔺适应重金属镉污染环境中的作用机制奠定基础。
[Abstract]:The long distance transport of CD mediated by heavy metal ATPase gene HMA2 plays an important role in CD distribution and detoxification in plants. However, the molecular mechanism of CD transport in Iris lactea remains unclear. In this study, heavy metal ATPase gene fragments were obtained by RT-PCR from total RNA in iris root. Based on the intermediate vector pH ANNIBAL and plant expression vector pART27, enzyme digestion and ligation were used. The RNAi plant expression vector pARTG1G2 driven by CaMV35S promoter containing the reverse repeat sequence of IlHMA2 gene fragment was constructed. The recombinant plasmid was introduced into Agrobacterium tumefaciens GV3101 by freeze-thaw method. This will lay a foundation for elucidating the mechanism of long distance transport of CD mediated by IlHMA2 in the adaptation of Iris marina to the environment polluted by heavy metal cadmium.
【作者单位】：山西农业大学林学院;北京市农林科学院北京草业与环境研究发展中心;
【基金】：北京市自然科学基金(6152008) 北京市农林科学院科技创新能力建设专项(KJCX20140103、KJCX20170110)
【分类号】：S688.4

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