Preparation of Cytolysin A (ClyA) Nanopores

Nicole Stéphanie Galenkamp, Veerle Van Meervelt, Natalie Lisa Mutter, Nieck Jordy van der Heide, Carsten Wloka, Giovanni Maglia*

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingChapterAcademicpeer-review

4 Citations (Scopus)


The ionic currents passing through nanopores can be used to sequence DNA and identify molecules at the single-molecule level. Recently, researchers have started using nanopores for the detection and analysis of proteins, providing a new platform for single-molecule enzymology studies and more efficient biomolecular sensing applications. For this approach, the homo-oligomeric Cytolysin A (ClyA) nanopore has been demonstrated as a powerful tool. Here, we describe a simple protocol allowing the production of ClyA nanopores. Monomers of ClyA are expressed in Escherichia coli and oligomerized in the presence of detergent. Subsequently, different oligomer variants are electrophoretically resolved and stored in a gel matrix for long-term use.

Original languageEnglish
Title of host publicationNanopore Technology
Subtitle of host publicationMethods in Molecular Biology
EditorsM.A. Fahie
Place of PublicationNew York
PublisherHumana Press
Number of pages8
ISBN (Electronic)978-1-0716-0806-7
ISBN (Print)978-1-0716-0805-0
Publication statusPublished - 2021

Publication series

NameMethods in Molecular Biology
ISSN (Print)1064-3745


  • Nanopores
  • Cytolysin A
  • ClyA
  • protein purification
  • protein assembly
  • pore-forming toxin
  • membrane protein
  • single-molecule
  • protein trapping
  • electrophysiology

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