水稻胚乳特异表达启动子DXCP35的克隆及功能鉴定
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国家重大科技专项 (2011ZX08001-001)


Cloning and functional indentification of an endosperm-specific promoter DXCP35 from rice
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    摘要:

    根据水稻全生育期基因表达谱芯片数据库找到1个在水稻胚乳特异表达的基因,命名为DX35,用PCR技术从水稻品种明恢63基因组中克隆得到其上游1 356 bp长度的启动子DXCP35。将DXCP35与β-glucuronidas(GUS)报告基因融合后,经根癌农杆菌(Agrobacterium tumefaciens)介导转化获得转基因水稻阳性植株,通过组织化学染色法证明DXCP35是1个胚乳特异表达启动子。对其进行5′端缺失分析,构建了6个缺失载体,通过验证缺失载体的表达谱,证明308 bp长度的启动子就足以维持胚乳特异表达模式。

    Abstract:

    Utilizing endosperm-specific promoter can effectively improve the quality of rice in genetic engineering,and has good application value in rice seed bioreactor.Based on microarray data,a rice endosperm-specific expression gene DX35 was identified.A promoter named as DXCP35 with 1 356 bp in length of the gene was cloned from the genomic DNA of Minghui 63 by PCR.Promoter DXCP35 was fused with β-glucuronidas(GUS) reporter gene and transformed into rice callus through Agrobacterium-mediated transformation.DXCP35 was identified as an endosperm-specific promoter in transgenic rice plants by histochemical staining.Results of analyzing its 5′ end deletion showed that promoter fragment with 308 bp length was sufficient to maintain the endosperm-specific expression model.The core function areas of DXCP35 were preliminarily identified.

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颜彦,林拥军.水稻胚乳特异表达启动子DXCP35的克隆及功能鉴定[J].华中农业大学学报,2014,33(05):15-20

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  • 收稿日期:2013-05-16
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