DNA Translocation Through Single Lysenin Channels
In this work, we exploited the use of a single lysenin channel as a sensor for characterization of ssDNA molecules by employing the resistive pulse technique. Lysenin creates a ~3 nm pore in a lipid bilayer, large enough for the translocation of macromolecules. We bathed a planar lipid membrane in 1M KCl solutions and inserted a single lysenin channel. Addition of 69 nucleotide ssDNA molecules into the reservoir situated at the cis side of the protein yielded multiple transients in the open current only when the electric field orientation was such that the DNA molecules were electrophoretically driven toward the nanopore. The analysis of the translocation events yielded a sharp distribution of the current blockages induced by the passing molecules. The dwell time distribution suggests strong interactions between the channel and ssDNA molecules. PCR performed on the translocated molecules followed by electrophoresis verified that the DNA molecules threads lysenin pores.
Belzeski, Philip, "DNA Translocation Through Single Lysenin Channels" (2017). 2017 Undergraduate Research and Scholarship Conference.