| 拟南芥 AtNHX5基因启动子的克隆和组织定位分析 |
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| 引用本文：李静雯，厚毅清，陈 军，朱天地.拟南芥 AtNHX5基因启动子的克隆和组织定位分析[J].西北农业学报,2019,28(12):2035~2042 |
| DOI:10.7606/j.issn.1004-1389.2019.12.016 |
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| 基金项目:国家自然科学基金（31660391，31460350）；甘肃省农业科学院农业科技自主创新专项现代生物育种项目（2019GAAS08）；甘肃省农业科学院中青年基金（2017GAAS92）。 |
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| 中文摘要:为了探明拟南芥内膜反向转运体 AtNHX5基因启动子（proNHX5）功能及基因的组织表达模式，从基因组中克隆 AtNHX5基因开放阅读框（ORF）上游侧翼调控区1 807 bp序列，发现该启动子具有典型启动子一般特征，不仅具有TATA-box、CAAT-box等核心元件，还含有与光响应、激素响应、逆境诱导响应和分生组织表达调控元件。构建 AtNHX5基因启动子与GUS的融合表达载体，通过农杆菌花序浸染法转化野生型拟南芥获得转基因植株。利用组织染色法鉴定转基因拟南芥的GUS表达模式，发现在子叶、下胚轴和花中有显著的GUS酶活性，成熟叶片和根中只有局部检测到GUS表达，在未成熟果荚中只有在果荚顶端和基部存在GUS酶活性，说明 AtNHX5基因启动子与GUS的融合表达载体成功构建且正常启动GUS基因表达，同时也表明， AtNHX5基因主要在这些部位表达，且表达具有时空特异性。 |
| 中文关键词: AtNHX5基因 启动子 克隆 GUS 表达分析 |
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| Molecular Cloning and Tissue Localization Analysis of AtNHX5 Gene Promoter from Arabidopsis thaliana |
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| Abstract:The endosomal AtNHX5 is one of the genes encoding Na+,K+/H+ antiporter in Arabidopsis thaliana.In order to test the function of proNHX5 and the tissue expression pattern of endosomal AtNHX5 gene,the sequence of proNHX5 from Arabidopsis thaliana Col-0,a 1 807 bp upstream of reading frame (ORF),was amplified by PCR.The analysis showed that proNHX5 had the general characteristics of typical promoter,which not only contained TATA-box and CAAT-box core components,but also contained control elements for light response,hormone response,adversity-induced response,and meristem expression.The fusion expression vector of proNHX5 and GUS gene was constructed and introduced into the Arabidopsis thaliana (ecotype Columbia 0) using the floral dip method.The expression pattern was monitored using GUS histochemical staining.As results.GUS staining was preferentially observed in cotyledons,hypocotyls and flowers of transgenic seedlings carrying the AtNHX5 promoter.In true leaves and roots,GUS activity was only partially detected for the AtNHX5 promoter.Whereas in the immature siliques,GUS staining was restricted to the silique tip and base.Collectively,these results suggested that fusion expression vector of AtNHX5 gene promoter and GUS was successfully constructed and it drived the expression of GUS gene successfully,and AtNHX5 gene predominantly expressed in these parts,and the expression had space-time specificity. |
| keywords: AtNHX5 gene Promoter Cloning GUS Expression analysis |
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