人BRD8启动子系列报告基因的构建和鉴定

    Construction and identification of serial human BRD8 reporter genes

    • 摘要: 目的: 构建具有相同3'末端而5'末端逐渐截短的人BRD8启动子系列报告基因,并进行鉴定。方法: 聚合酶链反应(PCR)从人基因组中扩增BRD8启动子片段并插入荧光素酶报告基因载体pGL3-Basic;以已构建的BRD8报告基因为模板,分别用PCR扩增具有相同3'末端而5'末端逐渐截短的BRD8启动子系列片段,并分别插入pGL3-Basic载体。对BRD8启动子系列报告基因进行酶切鉴定、测序。结果: 通过酶切鉴定、测序,证明构建的BRD8启动子系列报告基因序列和方向正确。结论: 成功构建了人BRD8启动子系列报告基因,为进一步研究调节BRD8基因表达的特异性转录因子和(或)沉默子奠定了基础。

       

      Abstract: Objective: To construct and identify serial human BRD8 luciferase reporter genes with the identical 3'-end but gradually truncated 5'-end. Methods: The BRD8 promotor fragment from human genome was amplified by polymerase chain reaction(PCR) and inserted into the luciferase reporter gene vector pGL3-Basic. A series of BRD8 promotor fragments with the identical 3'-end but gradually truncated 5'-end were produced by PCR using the above constructed BRD8 reporter gene vector as template and inserted into pGL3-Basic vector,respectively. The series of BRD8 reporter genes were identified by restriction enzyme digestion and sequencing. Results: The restriction enzyme digestion and DNA sequencing results showed that the sequences and orientation of the serial BRD8 reporter genes were correct. Conclusions: The serial human BRD8 reporter genes had been successfully constructed. These provide an experimental base for further research on the regulation of BRD8 gene transcription by some specific transcription factors and/or silencers.

       

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