22 results about "Pore Proteins" patented technology

Provided are a nanopore protein complex, a construction method therefor, and a use thereof. The nanopore complex comprises: a nanopore protein and an accessory protein. The nanopore protein comprises a nanopore cavity, and the nanopore cavity is formed by polymerizing a plurality of pore protein monomers. The accessory protein is formed by polymerizing a plurality of accessory protein monomers. The N-terminus of the accessory protein is embedded in the nanopore cavity and forms a continuous channel together with the nanopore cavity. On the basis of the moving direction of an analyte passing through the continuous channel, the continuous channel comprises a first sensing region and a second sensing region which are communicated sequentially, wherein the first sensing region is formed by part of the nanopore protein, and the second sensing region is formed by part or all of the accessory protein. A nanopore protein monomer is selected from a protein having an amino acid sequence as shown in SEQ ID NO: 1 and a variant thereof, and an accessory protein monomer is selected from a protein having an amino acid sequence as shown in SEQ ID NO: 3 and a variant thereof.

This invention belongs to the field of target characterization technology, and provides an application of PANX2 protein and its mutants as bioporins. The bioporins can detect one or more of the following: metal ions, inorganic salts, amino acids, nucleotides, small molecule drugs, small molecule diagnostic reagents, adenosine triphosphate (ATP), and monosaccharides. In this invention, one or more amino acids of the wild-type PANX2 protein are mutated to common amino acids other than the original amino acids. Compared to the wild-type PANX2 protein, the PANX2 protein mutants can be expressed normally, are stable, and show a significantly improved current signal during detection, with a longer amplitude and fewer spikes.

The invention provides a fusion method of pore protein and a membrane, a pore protein jack buffer solution and application. The fusion method comprises the following steps: distributing a first solution on the first side of the membrane, and distributing a second solution on the second side of the membrane; applying voltage to two sides of the membrane, fusing the pore protein and the membrane, and detecting current in the solution; when the current is increased, the pore protein is inserted into the membrane, the voltage is adjusted to 0V, and the fusion of the pore protein and the membrane is completed; the first solution comprises a first buffer solution containing pore protein, and the second solution comprises a second buffer solution without pore protein; the osmotic pressure of the first solution is greater than that of the second solution. The porous protein and membrane fusion single porosity problem in the prior art can be solved, and the method is suitable for the field of biological analysis and detection.

The present invention provides a nanopore protein complex and a use thereof. The nanopore protein complex comprises: a nanopore protein and multiple auxiliary proteins, the nanopore protein comprising multiple porin monomers and a nanopore channel structure formed by polymerization of the porin monomers, each auxiliary protein having an N-terminus thereof located within the nanopore channel structure, and the multiple auxiliary proteins being connected one-to-one with the multiple porin monomers and, together with the nanopore channel structure forming a continuous channel; along a direction in which an analyte passes through the continuous channel, the continuous channel comprises a first constriction region and a second constriction region that are connected in sequence, the first constriction region being formed at least in part by the nanopore protein, the second constriction region being formed in part or in whole by the auxiliary proteins, and the nanopore protein being wild-type or a mutant.

The application discloses application of a porin PorA in improving the ammonium nitrogen utilization capacity of a methanotrophic bacterium. The porin PorA has an amino acid sequence as shown in SEQ ID NO. 1. The coding gene porA of the porin PorA has a nucleotide sequence as shown in SEQ ID NO. 2. The application of the porin PorA in improving the ammonium nitrogen utilization capacity of Methylotuvimicrobium buryatense. Inactivation of the porin PorA can make the methanotrophic bacterium Methylotuvimicrobium buryatense grow by utilizing ammonium nitrogen, reduce the cost of fermenting the bacterium, and is of great significance for industrial application of the bacterium.

The application belongs to the technical field of methods for characterizing target polynucleotides, and provides an application of a biological nanopore protein and a mutant thereof in detection. The biological nanopore protein mutant is obtained by mutating a wild-type PANX1 nanopore protein, and an amino acid sequence of the wild-type PANX1 nanopore protein is shown as SEQ ID NO:1. The mutation refers to that one or more amino acids of the wild-type PANX1 nanopore protein are mutated into common amino acids other than original amino acids, or a C region of the wild-type PANX1 nanopore protein is truncated or truncated and replaced to obtain the PANX1 nanopore protein mutant. After the wild-type PANX1 nanopore protein is mutated, the wild-type PANX1 nanopore protein can be normally expressed. Compared with the wild-type PANX1 nanopore protein, the biological nanopore protein mutant is stable in property, and the C region is truncated to expand the pore diameter of the contraction region of the protein mutant, so that the pore channel is avoided from being blocked.

The present invention belongs to the technical field of characterization of target analyte properties, and particularly provides a mutant of a porin monomer, a protein pore comprising same, and use thereof in the detection of a target analyte, wherein an amino acid of the mutant of the porin monomer comprises a sequence set forth in SEQ ID NO: 1 or a sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 80%, 70%, 60%, or 50% identity thereto, and the amino acid of the mutant of the porin monomer comprises mutations at one or more positions corresponding to T71, S75, or F76 of SEQ ID NO: 1.

The invention relates to peptides and compounds that bind a two partner secretion (TPS) system protein. In some embodiments of the invention, the peptide or compound binds to a translocator pore protein (TpsB) thereby reducing translocation of a two-partner secretion exoprotein (TpsA) across a lipid membrane.

The application belongs to the technical field of biochemistry and molecular biology, and provides application of a two-component toxin nanopore protein XaxAB after site-directed mutation modification. The two-component toxin nanopore protein XaxAB comprises equal numbers of XaxA subunits and XaxB subunits, the amino acid sequence of the XaxA subunit is shown as SEQ ID NO: 1, and the amino acid sequence of the XaxB subunit is shown as SEQ ID NO: 2. The two-component toxin nanopore protein XaxAB is subjected to site-directed mutation modification, T290C mutation is introduced into the XaxA subunit and I30C mutation is introduced into the XaxB subunit, the two-component toxin nanopore protein XaxAB after site-directed mutation modification improves the current signal quality of pore opening, sequencing or detection, the current property is stable, the current signal width is narrower, and burrs are less.

The application belongs to the technical field of characterizing target samples, and provides application of enterococcus pore-forming toxin mutant protein in nanopore detection. The enterococcus pore-forming toxin mutant protein is an EPX1 nanopore mutant protein, which is obtained by mutation of wild-type EPX1 nanopore protein, and the amino acid sequence of the wild-type EPX1 nanopore protein is shown as SEQ ID NO: 1. The mutation refers to that one or more amino acids of the wild-type EPX1 nanopore protein are mutated into common amino acids other than the original amino acids. The EPX1 protein (wild type) of the application is a kind of pore-forming toxin protein, which widely exists in various organisms in nature. The mutant thereof exhibits excellent pore-forming ability after modification (after amino acid mutation). The EPX1 nanopore mutant protein realizes significant improvement in structural stability, molecular recognition efficiency and detection performance.

The present invention relates to novel Puncture protein monomers, Puncture protein pores formed from said monomers and the use of said Puncture protein pores in analyte detection and characterization.

Disclosed in the present invention are a nanopore protein monomer and a use thereof. The nanopore protein monomer is any one of the following polypeptides: (a1) a polypeptide having an amino acid sequence as set forth in SEQ ID NO: 4; (a2) a polypeptide derived from the amino acid sequence as set forth in SEQ ID NO: 4 by means of substitution and / or deletion and / or addition of one or more amino acids and having the same function; (a3) a polypeptide having 80% or above identity to the amino acid sequence defined in any one of (a1)-(a2) and having the same function; and (a4) a fusion polypeptide obtained after linking a tag to the terminus of the polypeptide defined in any one of (a1)-(a3). In the present invention, a nanopore protein which is stable and can be used for DNA detection is obtained on the basis of nanopores without a natural constriction region by a method of artificially designing a contraction region.

The present disclosure relates to relates methods and associated compositions that provide fast, efficient site-selective conjugation of a protein, such as the pore-forming protein α-hemolysin, to a biomolecule, such as a DNA polymerase, and the use of such site-selective protein-biomolecule conjugates in nanopore devices and methods.

The invention discloses a method for embedding nanopore protein into a biomimetic membrane and application of the biomimetic membrane. The method comprises the steps that the biomimetic membrane of a multi-block copolymer is prepared, and the multi-block copolymer contains a hydrophilic segment and a hydrophobic segment; embedding the nanopore protein into the biomimetic membrane; wherein the multi-block copolymer comprises a copolymer with two or more blocks; the biomimetic membrane contains an aromatic ring structure, and the aromatic ring structure is connected between the hydrophilic segment and the hydrophobic segment of the multi-block copolymer; and / or the nanopore protein contains an aromatic ring structure, and the aromatic ring structure is located on the amino acid, facing one side of the biomimetic membrane, of the transmembrane region of the nanopore protein. According to the method, the number of single holes and / or the single hole reserving rate during hole embedding of the biomimetic membrane can be increased.

This application provides a method and application for protecting nanoporins. The protection method includes: embedding the nanoporin onto a membrane, the membrane dividing a solution into two parts, which are connected through the nanoporin; and applying a negative voltage to one side of the solution containing the nanoporin before membrane crosslinking. This application enables an initiator to approach the membrane and undergo membrane crosslinking while remaining away from the nanoporin, thus protecting the nanoporin and preventing damage that could cause abnormalities in the opening current or even subsequent sequencing current.

A novel pore protein suitable for nanopore detection is provided, the pore protein is formed by polymerization of a pore protein monomer, the pore protein monomer comprises a cylinder segment, and the cylinder segment: a, has an amino acid sequence as shown in SEQ ID NO: 1; b, compared with the amino acid sequence as shown in SEQ ID NO: 1, the cylinder segment has an amino acid sequence formed by substitution, deletion and / or addition of one or more amino acids, and the cylinder segment has a function of forming a pore structure through polymerization; or c, compared with the amino acid sequence as shown in SEQ ID NO: 1, the cylinder section has an amino acid sequence with the identity of at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% and at least 99%, and the cylinder section has the function of forming a pore channel structure through polymerization.

The invention discloses a nanopore protein monomer and application thereof. The nanopore protein monomer is any one of the following polypeptides: (a1) a polypeptide with an amino acid sequence as shown in SEQ ID NO: 4; (a2) a polypeptide which is obtained by substitution and / or deletion and / or addition of one or more amino acids on the amino acid sequence as shown in SEQ ID NO: 4 and has the same function as the amino acid sequence as shown in SEQ ID NO: 4; (a3) a polypeptide which has more than 80% of identity with an amino acid sequence defined by any one of (a1)-(a2) and has the same function; and (a4) a fused polypeptide obtained by connecting a tag to the tail end of any polypeptide defined in (a1)-(a3). The stable nanopore protein capable of being used for DNA (deoxyribonucleic acid) detection is obtained by adopting an artificial constriction region design mode on the basis of a natural nanopore without a constriction region.

The application belongs to the technical field of characterizing target samples, and provides PANX3 biological pore protein, a mutant thereof and application of single molecule detection. The PANX3 biological pore protein is composed of seven identical subunits, the overall channel is funnel-shaped, and the amino acid sequence is shown as SEQ ID NO: 1. After one or more amino acids of the wild type PANX3 biological pore protein are mutated into common amino acids other than the original amino acids, compared with the wild type PANX3 biological pore protein, the mutant can be normally expressed and stable in property, and the current property and current signal of the mutant are obviously improved.

The present disclosure relates to compositions for the restoration of porosome function, comprising isolated porosome complexes that include a functional CFTR protein and a pharmaceutically acceptable excipient. These compositions are specifically formulated for inhaled or nasal administration, enabling delivery of the functional porosome complexes directly to the airway epithelium. By restoring the presence of functional CFTR within the porosome complex at the plasma membrane, the disclosed compositions address the underlying secretory defect in cystic fibrosis and provide a novel therapeutic approach for patients affected by this disease.

The present application provides a porin monomer, a porin, and a mutant and application thereof. The porin monomer includes: (a) a protein having an amino acid sequence shown in SEQ ID NO: 1; or (b) a protein mutant, where an amino acid sequence of the protein mutant is subjected to substitution, deletion and / or addition of one or more amino acids at at least one of the following sites in SEQ ID NO: 1: 77, 81, 82, and the like, and the mutant has a function of forming a pore channel structure by means of polymerization; or (c) a protein having at least 70%, 75%, 80%, 85%, 90%, 95%, or 99% homology to the protein in the (a) or the (b), where the mutant has the function of forming the pore channel structure by means of polymerization. Therefore, the problem in the prior art of a low capture rate of a nano porin to a library can be solved, and the present application is applicable to the field of single-molecule sequencing.

A novel pore protein suitable for nanopore detection is provided, the pore protein is formed by polymerization of a pore protein monomer, the pore protein monomer comprises a cylinder segment, and the cylinder segment: a, has an amino acid sequence as shown in SEQ ID NO: 1; b, compared with the amino acid sequence as shown in SEQ ID NO: 1, the cylinder segment has an amino acid sequence formed by substitution, deletion and / or addition of one or more amino acids, and the cylinder segment has a function of forming a pore structure through polymerization; or c, compared with the amino acid sequence as shown in SEQ ID NO: 1, the cylinder section has an amino acid sequence with the identity of at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% and at least 99%, and the cylinder section has the function of forming a pore channel structure through polymerization.

The invention relates to a novel HfaB nanopore protein mutant and application thereof in sequencing, and belongs to the technical field of nanopores. The polynucleotide for coding the nanopore protein mutant and the recombinant expression vector can express the nanopore protein mutant, and can be used for constructing recombinant cells or recombinant strains for expressing the nanopore protein mutant. The wild-type nanopore protein provided by the invention has relatively high pore stability and is suitable for nanopore sequencing, the sequencing current property of the nanopore protein mutant obtained by modification on the basis of the wild-type nanopore protein is further improved, the sequencing precision is obviously improved, the sequencing signal amplitude and step number are obviously increased, the pore current stability is enhanced, and the nanopore protein mutant can be applied to nanopore sequencing. The spontaneous blocking condition is reduced, and the abnormal noise on the via hole signal is reduced.