At2g04450启动子的克隆和启动活性分析
摘 要 为明确拟南芥At2g04450启动子对靶标基因表达的启动作用，根据已报道的拟南芥基因组序列设计合成1对引物，以拟南芥基因组DNA为模板克隆At2g04450上游调控序列，并与pBAR-GUS中间表达载体相连构建植物表达载体p04450p-GUS，采用农杆菌GV3101介导的渗透法转化野生型拟南芥，以GUS为报告基因研究该调控序列的组织表达特异性以及病原菌PstDC3000对该启动子的诱导表达效果。结果表明：获得At2g04450上游调控序列，该序列不仅具有启动子活性，并且具有组织特异性，GUS基因主要在根特异表达；病原菌PstDC3000对该启动子的表达没有诱导作用。本结果将为研究At2g04450的功能奠定基础。
关键词 At2g04450启动子；GUS活性；组织表达特异性
中图分类号 Q78 文献标识码 A
Molecular Cloning and Activity Analysis of a Promoter of
an Arabidopsis thaliana Gene At2g04450
WU Kunxin1，ZHANG Xiuchun1，CHEN Xiongting1，LIU Zhixin1，PENG Ming1，2*
1 Institute of Tropical Bioscience and Biotechnology，Chinese Academy of Tropical Agriculture Sciences/Key Laboratory
of Biology and Genetic Resources of Tropical Crops，Ministry of Agriculture，Haikou，Hainan 571101，China
2 Environmental Safety Supervision and Inspection Centre for Genetically Modified Plants and Microorganisms
used in Plants，Ministry of Agriculture，Haikou，Hainan 571101，China
Abstract 5′-UTR of At2g04450 gene was cloned by PCR from Arabidopsis thaliana. To investigate the tissue expression pattern and the possible response to pathogen infection of the cloned regulatory sequence，an expression vector containing this sequence fused with GUS was constructed for transformation into A. thaliana by using agrobacterium-mediated method. Histochemical staining of transgenic A. thaliana showed that GUS reporter gene was predominantly expressed in roots，and the activity of the promoter can not be induced by the PstDC3000. This research will facilitate the study of At2g044...
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