MSP nanopores and related methods
AssigneesUniversity of Washington, The UAB Research Foundation
InventorsJens H. Gundlach, Michael Niederweis, Thomas Z. Butler, Mikhail Pavlenok, Mark A. Troll, Suja Sukumaran
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Abstract
Provided herein are Mycobacterium smegmatis porin nanopores, systems that comprise these nanopores, and methods of using and making these nanopores. Such nanopores may be wild-type MspA porins, mutant MspA porins, wild-type MspA paralog porins, wild-type MspA homolog porins, mutant MspA paralog porins, mutant MspA homolog porins, or single-chain Msp porins. Also provided are bacterial strains capable of inducible Msp porin expression.
Claims28
A Mycobacterium smegmatis porin A (MspA) comprising an MspA monomer, wherein the MspA monomer comprises a variant of the sequence in SEQ ID NO:1, and wherein the variant comprises (i) mutations resulting in amino acids without a negative charge at positions 90 and 91 and (ii) one or more mutations adding a positively charged amino acid to a vestibule or entrance of the MspA.
A system comprising a Mycobacterium smegmatis porin (Msp) nanopore having a vestibule and a constriction zone that define a tunnel, wherein the tunnel is positioned between a first conductive liquid medium and a second conductive liquid medium allowing liquid communication between the first and second conductive liquid media, wherein the system is operative to detect an analyte when the system is subjected to an electric field sufficient to translocate the analyte from one conductive liquid medium to the other, and wherein the Msp is a mutant comprising at least a first mutant MspA monomer.
Dependencies are inferred from the claim text.