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首页> 外文期刊>African Journal of Biotechnology >Analysis of gene expression by promoter reporter constructs and knockout plants: A case study under phosphorus stress
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Analysis of gene expression by promoter reporter constructs and knockout plants: A case study under phosphorus stress

机译：启动子报告基因构建体和敲除植物的基因表达分析：以磷胁迫为例

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Promoter reporter constructs were developed using Gateway Cloning Technology for plant non-specific lipid transport proteins (nsLTP1,?AT4g12545 and nsLTP2, AT5g46900), MADS-Box (AT4g37940) and?leucine rich repeat (LRR,?AT2g28970)genes. The purpose of the construct was to investigate gene expression and function under phosphorus (P) stress.?Agrobacterium tumefaciens?was used to transform?Arabidopsis thaliana. Transformed (T1), knockout and wild type ecotype Colombia-0?A. thaliana?were investigated. To monitor the gene activity, plants were grown under different conditions: fully inside solid media, on the surface of solid media and hydroponically with 0.5 mM l-1?phosphorus and without phosphorus. Plants were also grown on MS media with selectable marker containing?50?mM l-1Kanamycin.?Morphological root assessment was also done by using knockoutplants?for the respective promoters. The promoters under study were induced promoter. GUS staining was observed on T1?plants when planted fully inside solid media and on the selectable marker for all the constructs regardless of phosphorus treatments. Mechanical impedance, oxygen deficiency and stress due to selectable marker were the probable reasons for the root specific gene expression. This suggests that these genes were involved in adaptation mechanism for plants under such stress conditions.
机译：使用Gateway Cloning Technology开发了启动子报告基因构建体，用于植物非特异性脂质转运蛋白（nsLTP1，？AT4g12545和nsLTP2，AT5g46900），MADS-Box（AT4g37940）和富含亮氨酸的重复序列（LRR，？AT2g28970）基因。该构建体的目的是研究在磷（P）胁迫下的基因表达和功能。根癌农杆菌用于转化拟南芥。转化（T1），基因敲除和野生型生态型Columbia-0？A。拟南芥。为了监测基因活性，使植物在不同条件下生长：完全在固体培养基中，在固体培养基的表面上，并与0.5mM的l-1β磷无磷地进行水培。还使植物在具有选择标记的MS培养基上生长，该选择标记含有〜50μmM-1-1卡那霉素。形态学根系评估也通过使用敲除植物对各个启动子进行。研究中的启动子是诱导型启动子。当完全种植在固体培养基中时，无论磷处理如何，在T1植物上均观察到GUS染色，并且在所有构建体的选择标记上均观察到GUS染色。机械阻抗，缺氧和因选择标记引起的胁迫是根特异性基因表达的可能原因。这表明这些基因参与了这种胁迫条件下植物的适应机制。

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