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Single-walled carbon nanotubes as scaffolds to concentrate DNA for the study of DNA-protein interactions.

Authors :
Liu Z
Galli F
Waterreus WJ
Meulenbroek E
Koning RI
Lamers GE
Olsthoorn RC
Pannu N
Oosterkamp TH
Koster AJ
Dame RT
Abrahams JP
Source :
Chemphyschem : a European journal of chemical physics and physical chemistry [Chemphyschem] 2012 Apr 23; Vol. 13 (6), pp. 1569-75. Date of Electronic Publication: 2012 Mar 07.
Publication Year :
2012

Abstract

Genomic DNA in bacteria exists in a condensed state, which exhibits different biochemical and biophysical properties from a dilute solution. DNA was concentrated on streptavidin-covered single-walled carbon nanotubes (Strep-SWNTs) through biotin-streptavidin interactions. We reasoned that confining DNA within a defined space through mechanical constraints, rather than by manipulating buffer conditions, would more closely resemble physiological conditions. By ensuring a high streptavidin loading on SWNTs of about 1 streptavidin tetramer per 4 nm of SWNT, we were able to achieve dense DNA binding. DNA is bound to Strep-SWNTs at a tunable density and up to as high as 0.5 mg mL(-1) in solution and 29 mg mL(-1) on a 2D surface. This platform allows us to observe the aggregation behavior of DNA at high concentrations and the counteracting effects of HU protein (a histone-like protein from Escherichia coli strain U93) on the DNA aggregates. This provides an in vitro model for studying DNA-DNA and DNA-protein interactions at a high DNA concentration.<br /> (Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Details

Language :
English
ISSN :
1439-7641
Volume :
13
Issue :
6
Database :
MEDLINE
Journal :
Chemphyschem : a European journal of chemical physics and physical chemistry
Publication Type :
Academic Journal
Accession number :
22407519
Full Text :
https://doi.org/10.1002/cphc.201100896