TY - JOUR
T1 - In vitro footprinting of promoter regions within supercoiled plasmid DNA.
AU - Sun, Daekyu
PY - 2010
Y1 - 2010
N2 - 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 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.
AB - 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 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.
UR - http://www.scopus.com/inward/record.url?scp=77449137206&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=77449137206&partnerID=8YFLogxK
U2 - 10.1007/978-1-60327-418-0_14
DO - 10.1007/978-1-60327-418-0_14
M3 - Article
C2 - 19997887
AN - SCOPUS:77449137206
SN - 1064-3745
VL - 613
SP - 223
EP - 233
JO - Methods in molecular biology (Clifton, N.J.)
JF - Methods in molecular biology (Clifton, N.J.)
ER -