In vitro footprinting of promoter regions within supercoiled plasmid DNA.
In vitro footprinting of promoter regions within supercoiled plasmid DNA.
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DOI：
10.1007/978-1-60327-418-0_14
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发表时间：
2010
期刊：
影响因子：
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通讯作者：
Sun, Daekyu
中科院分区：
文献类型：
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作者：
Sun, Daekyu
多聚嘌呤/多聚嘧啶（pPU/pPY）片段存在于许多生长相关基因的启动子区域，其构象具有很强的动态性。在这一章中，我们描述了一个详细的协议DNase I和S1核酸酶足迹实验与超螺旋质粒DNA含有这样的启动子区域，以探测是否有构象转换到B型DNA，熔化的DNA和G-四链体结构在这道。这与人血管内皮生长因子（VEGF）基因的近端启动子区，其中还包含多个结合位点的Sp1和Egr-1转录因子证明。
Polypurine/polypyrimidine (pPU/pPY) tracts, which exist in the promoter regions of many growth-related genes, have been proposed to be very dynamic in their conformation. In this chapter, we describe a detailed protocol for DNase I and S1 nuclease footprinting experiments with supercoiled plasmid DNA containing such the promoter regions to probe whether there are conformational transitions to B-type DNA, melted DNA and G-quadruplex structures within this tract. This is demonstrated with the proximal promoter region of the human vascular endothelial growth factor (VEGF) gene, which also contains multiple binding sites for Sp1 and Egr-1 transcription factors.