Preparation of Fragaceatoxin C (FraC) Nanopores

Natalie Lisa Mutter, Gang Huang, Nieck Jordy van der Heide, Florian Leonardus Rudolfus Lucas, Nicole Stéphanie Galenkamp, Giovanni Maglia, Carsten Wloka*

*Bijbehorende auteur voor dit werk

Onderzoeksoutput: ChapterAcademicpeer review

Samenvatting

Biological nanopores are an emerging class of biosensors with high-end precision owing to their reproducible fabrication at the nanometer scale. Most notably, nanopore-based DNA sequencing applications are currently being commercialized, while nanopore-based proteomics may become a reality in the near future.Although membrane proteins often prove to be difficult to purify, we describe a straightforward protocol for the preparation of Fragaceatoxin C (FraC) nanopores, which may have applications for DNA analysis and nanopore-based proteomics. Recombinantly expressed FraC nanopores are purified via two rounds of Ni-NTA affinity chromatography before and after oligomerization on sphingomyelin-containing liposomes. Starting from a plasmid vector containing the FraC gene, our method allows the production of purified nanopores within a week. Afterward, the FraC nanopores can be stored at +4 °C for several months, or frozen.

Originele taal-2English
TitelNanopore Technology
SubtitelMethods in Molecular Biology
RedacteurenM.A. Fahie
Plaats van productieNew York
UitgeverijHumana Press
Pagina's3-10
Aantal pagina's8
ISBN van elektronische versie978-1-0716-0806-7
ISBN van geprinte versie978-1-0716-0805-0
DOI's
StatusPublished - 2021

Publicatie series

NaamMethods in Molecular Biology
Volume2186
ISSN van geprinte versie1064-3745

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