The Expression of α-Hemolysin and the Preparation of Its Nanopore
ZHUO Li-xia;WANG Ying;ZHANG Chun-ping;DUAN Jing;ZHANG Ya-ni;The College of Life Science,Northwest University;College of Chemistry & Mateirals Secience,Northwest University;
α-hemolysin( αHL) is a channel-forming toxin released by Staphylococcus aureus,which can form a transmembrane heptamers nanopore on eukaryotic cell membranes and other lipid bilayers. The αHL nanopore has been popularly used as a single molecule biosensor. In the present study,αHL was expressed in the host strain BL21( DE3) p Lys S and purified by using molecular weight cut-off of ultrafiltration membranes. αHL heptameric nanopore was formed on rabbit red blood cell memberanes and characterized by using patch clamp technique and single channel current recording. αHL heptameric nanopore showed a stable structure and good ion permeability on lipid bilayer. It can be used as a stable single-molecule detection device. These results make a useful work for the preparation of α-hemolysin heptameric nanopore and the further exploring in the filed of single-molecule detection.