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.
Claims21
A nucleic acid comprising a sequence encoding a Mycobacterium smegmatis porin A (MspA) monomer, wherein the encoded MspA monomer comprises a variant amino acid sequence of SEQ ID NO:1, the variant amino acid sequence comprising:
- (i) mutations resulting in amino acids with a neutral charge at each of positions 90 and 91, wherein the mutations at positions 90 and 91 are each individually selected from asparagine, glutamine, or tyrosine;
- and
- (ii) one or more mutations replacing a negatively charged amino acid in SEQ ID NO:1 with a positively charged amino acid, thereby adding one or more positively charged amino acid to a vestibule or entrance of an MspA comprising the encoded MspA monomer.
A nucleic acid comprising a sequence encoding a Mycobacterium smegmatis porin A (MspA) monomer, wherein the encoded MspA monomer comprises a variant amino acid sequence of SEQ ID NO:1, and wherein the variant amino acid sequence comprises (i) mutations resulting in amino acids without a negative charge at positions 90 and 91 and (ii) one or more mutations replacing a negatively charged amino acid with a positively charged amino acid at one or more of amino acid positions 118, 134, and 139.
Dependencies are inferred from the claim text.