title: Creating a Single Sensing Zone within an Alpha-Hemolysin Pore Via Site Directed Mutagenesis.
abstract: Although significant progress has recently been made towards realizing the goal of direct nanopore based DNA sequencing [1], there are still numerous hurdles that need to be overcome. One such hurdle associated with the use of the biological nanopore α-hemolysin (αHL) is the fact that the wild type channel contains three very distinct recognition or sensing regions within the β-barrel [2, 3], making identification of the bases residing within or moving through the pore very difficult. Through site directed mutagenesis, we have been able to selectively remove one of two sensing regions while simultaneously enhancing the third. Our approach has led to the creation of αHL pores containing single sensing zones and provides the basis for engineering αHL pores suitable for direct DNA sequencing.
authors.affiliation: Electronic BioSciences, Inc., 5754 Pacific Center Blvd. Suite 204, San Diego, CA 92121.
authors.forename: Eric N
authors.initials: EN
authors.lastname: Ervin
authors.affiliation: Electronic BioSciences, Inc., 5754 Pacific Center Blvd. Suite 204, San Diego, CA 92121.
authors.forename: Geoffrey A
authors.initials: GA
authors.lastname: Barrall
authors.affiliation: Electronic BioSciences, Inc., 5754 Pacific Center Blvd. Suite 204, San Diego, CA 92121.
authors.forename: Prithwish
authors.initials: P
authors.lastname: Pal
authors.affiliation: Electronic BioSciences, Inc., 5754 Pacific Center Blvd. Suite 204, San Diego, CA 92121.
authors.forename: Megan K
authors.initials: MK
authors.lastname: Bean
authors.affiliation: Electronic BioSciences, Inc., 5754 Pacific Center Blvd. Suite 204, San Diego, CA 92121.
authors.forename: Anna E P
authors.initials: AE
authors.lastname: Schibel
authors.affiliation: Electronic BioSciences, Inc., 5754 Pacific Center Blvd. Suite 204, San Diego, CA 92121.
authors.forename: Andrew D
authors.initials: AD
authors.lastname: Hibbs
keywords: DNA sequencing; Nanopore; alpha-Hemolysin; sensing zone; site directed mutagenesis