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转录因子CREB对合浦珠母贝无定型碳酸钙结合蛋白启动子的调节作用

作  者:
杨易;高静;陈彦;鲁冰;谢莉萍;张荣庆
单  位:
清华大学生命科学学院
关键词:
合浦珠母贝;无定型碳酸钙结合蛋白;环腺苷效应元件结合蛋白;启动子;转录活性
摘  要:
【目的】研究无定型碳酸钙结合蛋白(ACCBP)基因的上游调控机制。【方法】以合浦珠母贝(Pinctada fucata)为研究对象,利用基因组步移技术克隆合浦珠母贝ACCBP基因(PfACCBP)的启动子。设计启动子截短实验,通过双荧光素酶标记法确定启动子与转录因子CREB的结合位点,定位启动子中的功能元件。克隆转录因子环腺苷效应元件结合蛋白(CREB),并通过共转染实验和凝胶迁移率分析检测启动子的相对转录活性。【结果】获得合浦珠母贝ACCBP基因的启动子序列,ACCBP启动子在HEK-293T细胞系中转录活性较高;该启动子上存在CRE元件,该元件可在转录过程中与CREB蛋白直接结合,大幅上调ACCBP启动子的转录效率。【结论】转录因子CREB可通过与ACCBP启动子上的CRE元件结合而增强转录活性的调控作用。
译  名:
Regulation of ACCBP Promoter by Transcription Factor CREB in Pinctada fucata
作  者:
YANG Yi;GAO Jing;CHEN Yan;LU Bing;XIE Li-ping;ZHANG Rong-qing;School of Life Science, Tsinghua University;
关键词:
Pinctada fucata;;Amorphous Calcium Carbonate Binding Protein;;cAMP-response element binding protein;;promoter;;transcription ability
摘  要:
【Objective】To study the promoter region in the regulatory mechanism of ACCBP.【Method】The promoter of ACCBP were cloned by genome-walking PCR. Promoter deletion analysis clones were produced for the dual-luciferase reporter assay to find the binding site of transcription factor CREB, and locate the functional element of the promoter. A classic transcription factor, cAMP-response element binding protein(CREB), was cloned for transient co-transfection assays and electrophoretic mobility shift assay for detecting the relative transcription activity of the promoter. 【Result】The promoter of ACCBP was sequenced, and the promoter showed high transcription activity in cell line HEK-239 T. The promoter was confirmed to have a CRE element that interacted with CREB and resulted in enhancing the transcription efficiency 【Conclusion】CREB can enhance the transcription ability of ACCBP by binding the CRE site on the promoter.

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