Preparation of Cytolysin A (ClyA) NanoporesGalenkamp, N. S., Van Meervelt, V., Mutter, N. L., van der Heide, N. J., Wloka, C. & Maglia, G., 2021, Nanopore Technology: Methods in Molecular Biology. Fahie, M. A. (ed.). New York: Humana Press, p. 11-18 8 p. (Methods in Molecular Biology; vol. 2186).
Research output: Chapter in Book/Report/Conference proceeding › Chapter › Academic › peer-review
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.
|Title of host publication||Nanopore Technology|
|Subtitle of host publication||Methods in Molecular Biology|
|Place of Publication||New York|
|Number of pages||8|
|Publication status||Published - 2021|
|Name||Methods in Molecular Biology|
- Nanopores, Cytolysin A, ClyA, protein purification, protein assembly, pore-forming toxin, membrane protein, single-molecule, protein trapping, electrophysiology