Platform for in situ real-time measurement of protein-induced conformational changes of DNA.
A platform for in situ and real-time measurement of protein-induced conformational changes in dsDNA is presented. We combine electrical orientation of surface-bound dsDNA probes with an optical technique to measure the kinetics of DNA conformational changes. The sequence-specific Escherichia coli integration host factor is utilized to demonstrate protein-induced bending upon binding of integration host factor to dsDNA probes. The effects of probe surface density on binding/bending kinetics are investigated. The platform can accommodate individual spots of microarrayed dsDNA on individually controlled, lithographically designed electrodes, making it amenable for use as a high throughput assay.
Spuhler
Philipp S
PS
Department of Biomedical Engineering, Boston University, Boston, MA 02215, USA.
Knezevic
Jelena
J
Yalçin
Ayça
A
Bao
Qiuye
Q
Pringsheim
Erika
E
Dröge
Peter
P
Rant
Ulrich
U
Unlü
M Selim
MS
D015342:DNA Probes / Q000032:analysis* / Q000737:chemistry; D004269:DNA, Bacterial / Q000032:analysis* / Q000737:chemistry; D004566:Electrodes; D004926:Escherichia coli / Q000737:chemistry*; D029968:Escherichia coli Proteins / Q000032:analysis* / Q000737:chemistry; D006046:Gold / Q000737:chemistry; D009690:Nucleic Acid Conformation; D013050:Spectrometry, Fluorescence; D013997:Time Factors
D015342:DNA Probes; D004269:DNA, Bacterial; D029968:Escherichia coli Proteins; D006046:Gold