水稻胚特异表达基因Os08PTS启动子的克隆及分析

    Cloning and Analysis of the Promoter of Rice Embryo-specific Expression Gene Os08PTS

    • 摘要: 为分析编码磷脂酰肌醇转移蛋白基因Os08PTS在水稻种子发育中的功能及利用Os08PTS启动子进行遗传改良。依据T-DNA插入位点信息,克隆Os08PTS基因启动子,并利用生物信息学手段分析Os08PTS启动子的顺式作用元件特征,同时构建启动子与GUS融合表达载体,通过农杆菌介导法获得转基因植株,验证启动子的表达特征。结果表明:Os08PTS基因启动子片段中包含启动子的基本元件TATA-box和CAAT-box,以及光调控元件、赤霉素应答元件、生长素反应元件、参与防御和应激反应相关元件等多个与生长发育或者逆境胁迫相关的顺式作用元件,还含有与分生组织表达以及胚乳表达相关元件等。进一步构建Os08PTS启动子的GUS融合载体,并获得水稻遗传转化植株。Os08PTS启动子序列能够驱动Os08PTS基因在水稻茎和种子胚的表达,该启动子具有驱动下游基因转录的活性。

       

      Abstract: In order to analyze the function of Os08PTS gene encoding the phosphatidylinositol transfer protein in rice seed development and to use Os08PTS promoter for genetic improvement, the promoter of Os08PTS gene was cloned by using T-DNA insertion site information, and the cis acting element characteristics of Os08PTS promoter were analyzed by using the bioinformatics methods. At the same time, the fusion expression vector of the promoter and GUS was constructed, and the transgenic plants were obtained by using the agrobacterium-mediated method to verify the expression characteristics of the promoter. The results showed that the promoter fragment of Os08PTS gene contained the basic elements of the promoter, TATA-box and CAAT-box, and several cis acting elements related to growth and development or stress response, such as the light regulatory elements, gibberellin responsive elements, auxin response elements, elements involved in defense and stress response, etc, as well as the elements related to meristem expression and endosperm expression. The GUS fusion vector of Os08PTS promoter was constructed and the rice genetic transformation plants were obtained. Os08PTS promoter sequence could drive the expression of Os08PTS gene in rice stem and seed embryo, and the promoter had the activity of driving the downstream gene transcription.

       

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