Naslov
Single Molecule Localization and Discrimination of DNA–Protein Complexes by Controlled Translocation Through Nanocapillaries

Autori
Bulushev, Roman D. ; Marion, Sanjin ; Petrova, Ekaterina ; Davis, Sebastian J. ; Maerkl, Sebastian J. ; Radenovic, Aleksandra

Izvornik
Nano letters (1530-6984) 16 (2016), 12; 7882-7890

Vrsta, podvrsta i kategorija rada
Radovi u časopisima, članak, znanstveni

Ključne riječi
DNA−protein complex ; force measurements ; Jarzynski equation ; nanocapillary ; Nanopore ; protein binding site

Sažetak
Through the use of optical tweezers we performed controlled translocations of DNA– protein complexes through nanocapillaries. We used RNA polymerase (RNAP) with two binding sites on a 7.2 kbp DNA fragment and a dCas9 protein tailored to have five binding sites on λ-DNA (48.5 kbp). Measured localization of binding sites showed a shift from the expected positions on the DNA that we explained using both analytical fitting and a stochastic model. From the measured force versus stage curves we extracted the nonequilibrium work done during the translocation of a DNA–protein complex and used it to obtain an estimate of the effective charge of the complex. In combination with conductivity measurements, we provided a proof of concept for discrimination between different DNA–protein complexes simultaneous to the localization of their binding sites.

Izvorni jezik
Engleski

Znanstvena područja
Fizika, Biologija

POVEZANOST RADA