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Nucleic Acids Res ; 41(20): e189, 2013 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-24003032

RESUMO

The effect of a cationic-neutral diblock polypeptide on the conformation of single DNA molecules confined in rectangular nanochannels is investigated with fluorescence microscopy. An enhanced stretch along the channel is observed with increased binding of the cationic block of the polypeptide to DNA. A maximum stretch of 85% of the contour length can be achieved inside a channel with a cross-sectional diameter of 200 nm and at a 2-fold excess of polypeptide with respect to DNA charge. With site-specific fluorescence labelling, it is demonstrated that this maximum stretch is sufficient to map large-scale genomic organization. Monte Carlo computer simulation shows that the amplification of the stretch inside the nanochannels is owing to an increase in bending rigidity and thickness of bottlebrush-coated DNA. The persistence lengths and widths deduced from the nanochannel data agree with what has been estimated from the analysis of atomic force microscopy images of dried complexes on silica.


Assuntos
DNA/química , Peptídeos/química , Mapeamento Cromossômico , DNA/ultraestrutura , Técnicas Analíticas Microfluídicas/instrumentação , Microscopia de Força Atômica , Microscopia de Fluorescência , Método de Monte Carlo , Nanoestruturas/química
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