84 results about "Prokaryotic cells" patented technology

The invention relates to the improvement of endonuclease-based antimicrobials by blocking DNA repair of double-strand break(s) (DSB(s)) in prokaryotic cells. In this respect, the invention especially concerns a method involving blocking DNA repair after a nucleic acid has been submitted to DSB, in particular by a Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) associated programmable double-strand endonuclease. The invention particularly relates to the use of an exogenous molecule that inhibits DNA repair, preferably a protein that binds to the ends of the double-stranded break to block DSB repair. The invention also relates to vectors, particularly phagemids and plasmids, comprising nucleic acids encoding nucleases and Gam proteins, and a pharmaceutical composition and a product containing these vectors and their application.

The invention relates to a genetically engineered bacterium for producing a resuscitation promoting factor Rpf and application of the genetically engineered bacterium, and belongs to the technical field of bioengineering. The genetically engineered bacterium is a prokaryotic cell, and the nucleotide sequence of the resuscitation promoting factor Rpf is as shown in SEQ. ID. NO.1. The genetically engineered bacterium is preserved in China Center for Type Culture Collection. The Rpf recombinant protein prepared from the genetically engineered bacterium constructed by the invention is in soluble expression, and a good research foundation is laid for subsequent detection of VBNC state pathogenic bacteria. After the recombinant protein is purified through affinity chromatography, the purity of the Rpf recombinant protein can reach 93% and is far higher than the extraction purity in the prior art.

The invention discloses a human-derived anti-respiratory syncytial virus neutralizing antibody RS135 and application of the human-derived anti-respiratory syncytial virus neutralizing antibody RS135. The human neutralizing antibody specifically aiming at the F protein of the respiratory syncytial virus is successfully obtained by applying a phage antibody library technology; the obtained human neutralizing anti-respiratory syncytial virus F protein gene engineering antibody variable region gene, Fab antibody gene and full antibody gene under the characteristics of each antibody gene are utilized to obtain the human neutralizing anti-respiratory syncytial virus F protein gene engineering antibody. The antibody can be expressed and produced in prokaryotic cells, yeast cells, eukaryotic cells and any recombination system or any other reconstructed gene containing the antibody gene on the basis of the antibody, and an antibody product capable of neutralizing respiratory syncytial virus infection is obtained. And a specific antibody drug for clinically preventing and treating respiratory syncytial virus infection is prepared.

The invention relates to the technical field of nucleic acid detection, in particular to an integrated consumable device and method integrating nucleic acid sample preparation and volume metering. The sample treatment tube filled with a liquid sample is butted to the mounting ring, and then the liquid sample in the sample treatment tube is heated by the heater, so that enough high temperature is generated in the sample treatment tube to lyse eukaryotic cells or prokaryotic cells or viruses and the like in the sample and release nucleic acid. The temperature can also be high enough to denature proteins and inactivate pathogens, and the elevated temperature can create a sufficient internal gas pressure sufficient to enter a liquid sample through the connection trough and the three-way valve into the metering container, the liquid sample can fill the space of the metering container, the liquid will contact the actuator to activate the actuator, and the liquid will enter the metering container through the connection trough and the three-way valve. The three-way valve is indexed to close liquid outflow of the sample treatment tube, and a channel between the metering container and the reaction tube is opened at the same time, so that the metered sample is moved into the reaction tube connected with the butt joint ring.

The invention provides an anti-influenza B virus nano antibody and application thereof, the nano antibody comprises a heavy chain variable region, the heavy chain variable region comprises CDR1, CDR2 and CDR3, and the CDR3 of the heavy chain variable region is selected from an amino acid sequence as shown in SEQ ID NO: 8 or SEQ ID NO: 16. The nano antibody is obtained through a multi-antigen cross immunization and panning method, the nano antibody can be specifically combined with Colorado / 06 / 2017 and Florida / 4 / 2006 influenza B viruses, the sensitivity is high, the specificity is high, and the phenomenon of missing detection in clinical detection can be avoided in clinical application. The nano antibody can be efficiently expressed through prokaryotic cells, and compared with eukaryotic mammalian cell expression of a traditional antibody, the nano antibody is lower in production cost and more beneficial to popularization and application in detection of infectious diseases.

The invention relates to a method for preparing a sample for microscopic examination comprising the steps of using a chitosan-based porous material having interconnected pores with sizes between 5 μm and 700 μm and a total porosity (volumetric fraction) between 40% and 90%, adding at least one biopsy sample containing tissue and / or eukaryotic and / or prokaryotic cells and / or eukaryotic cells containing viral particles to the chitosan-based porous material; and adding (C1-C6)-alkyl alcohol or mixtures thereof.

The present invention relates to a device for the collection, pre-analytical treatment, transport and / or grinding of a solid sample, comprising a flask for receiving the solid sample, a pre-analytical medium comprised in the flask and a plurality of beads also comprised in the flask, each bead having a diameter equal to or greater than 5 mm and a density of at least 5.0 g.cm-3. The present invention also relates to the use of said device and to a method for extracting all or part of at least one eukaryotic or prokaryotic cell from a solid sample.

An IgG1 Fc monomer, a preparation method therefor and an application thereof. Causing an Fc dimer of a novel IgG1 Fc monomer sequence for which an antibody IgG1 constant region is modified to become an Fc monomer by means of the modification on a human antibody IgG1 constant region Fc that uses antibody engineering technology, and maintaining an FcRn binding function; the present application has the feature of very low non-specific binding with unrelated proteins, and the main features of the Fc monomer comprise the T366, L368, P395, and K409 positions in a CH3 region of the constant region of the antibody having mutations, and the monomer being highly efficiently expressed in prokaryotic cells; the monomer may bind to FcRn by using a pH-dependent specific binding mode, and has the feature of very low non-specific binding. By using said Fc monomer, the same may be fused or coupled with various proteins, polypeptides, small molecules, nucleic acids and the like for different targets such that the fused or coupled molecules have the feature of being capable of binding to FcRn by using pH dependence.

Variant or mutant prokaryotic cells are disclosed that express elevated or increased levels of a wide variety of active proteins. Also disclosed are methods of producing the elevated or increased levels of active encoded proteins.

To provide, generally, methods and compositions for transferring a genetic circuit from one prokaryotic cell ("donor cell") to another prokaryotic cell ("recipient cell" or "target cell" used interchangeably herein).SOLUTION: The present disclosure relates to: prokaryotic donor cells comprising (i) a genetic circuit of interest and (ii) one or more expressed transcriptional repressor proteins; and the use of the donor cells in efficient transfer of the genetic circuit into a prokaryotic recipient cell. The genetic circuit includes nucleic acid sequences encoding an RNA molecule or protein of interest.SELECTED DRAWING: None

Methods for producing organosulfur compounds from a prokaryotic cell comprising enzymes involved in the production of cysteamine, hypotaurine, and taurine. Said prokaryote may comprise a vanin selected from vanin-1 (vnn-1), vanin-2 (vnn-2), or vanin- 3 (vnn-3), a cysteamine dioxygenase (ado), and / or a flavin-containing monooxygenase 1 (fmo1). Methods include the expression of organosulfur compounds in Corynebacterium glutamicum with incubation conditions to promote compound production.

The present application belongs to the technical field of genetic engineering, and particularly relates to a newly discovered micro DNA nuclease, which can realize recognition, combination and cutting of specific DNA sequences under the guidance of specific gRNA sequences. The nuclease N3 and N4 which can recognize new PAM sequences and have smaller volume are obtained through the present application, and the editing effect and characteristics of the nuclease in prokaryotic cells and eukaryotic cells are verified, which is expected to expand the application scenarios in the fields of gene editing and gene expression regulation.

Disclosed are methods, systems, components, and compositions for cell-free synthesis of glycosylated carrier proteins. The glycosylated carrier proteins may be utilized in vaccines, including anti-bacterial vaccines. The glycosylated carrier proteins may include a bacterial polysaccharide conjugated to a carrier, which may be utilized to generate an immune response in an immunized host against the polysaccharide conjugated to the carrier. The glycosylated carrier proteins may be synthesized in cell-free glycoprotein synthesis (CFGpS) systems using prokaryote cell lysates that are enriched in components for glycoprotein synthesis such as oligosaccharyltransferases (OSTs) and lipid-linked oligosaccharides (LLOs) including OSTs and LLOs associated with synthesis of bacterial O antigens.

A genetically modified prokaryotic cell to express enzymes involved in the production of taurine. Said prokaryote comprises a vanin selected from: vanin- 1 (vnn-1), vanin-2 (vnn-2), or vanin- 3 (vnn-3), a cysteamine dioxygenase (ado), and a flavin-containing monooxygenase 1 (fmol), in addition to either the addition, alteration, or deletion of at least one gene involved in taurine synthesis or transport.

The invention relates to a method for improving the prokaryotic expression quantity of CRISPR-dCas9 (clustered regularly interspaced short palindromic repeats) protein. Specifically, the invention relates to a method for improving the expression level of dCas9 protein in a prokaryotic cell, and a leucine zipper gene sequence and a dCas9 gene sequence are connected and introduced into the prokaryotic cell. According to the method, the high-purity and high-concentration dCas9 protein can be obtained, the expression quantity of the dCas9 protein is remarkably higher than that of a common method using maltose binding protein (MBP) as a fusion molecular chaperone, the obtained dCas9-lzip fusion protein still has the capability of being normally combined with DNA in a targeted manner under the guidance of sgRNA, and the function of dCas9 is not influenced by connection of a lzip sequence.

To provide an expression and secretion system, and methods of using the same, for the expression and secretion of one fusion protein in prokaryotic cells and a second fusion protein in eukaryotic cells.SOLUTION: Provided herein are nucleic acid molecules, vectors and host cells comprising such vectors and nucleic acid molecules. The nucleic acid molecule encodes a first polypeptide comprising VH-HVR1, VH-HVR2 and VH-HVR3 of a variable heavy chain domain (VH) and / or a second polypeptide comprising VL-HVR1, VL-HVR2 and VL-HVR3 of a variable light chain domain (VL). The nucleic acid molecule further encodes a signal sequence which is functional in both prokaryotic and eukaryotic cells and is encoded by a nucleic acid sequence that is operably linked to the first and / or second polypeptide sequence. A full-length antibody is expressed from the first and / or second polypeptide of the nucleic acid molecule.SELECTED DRAWING: Figure 14

A genetically modified prokaryotic cell comprising:at least one of the following:i. an addition, deletion and / or alteration of at least one gene to promote to taurine production; andii. an addition, deletion and / or alteration of at least one gene related to taurine cellular transportation; andat least one of the following polynucleotide sequences:i. a vanin (vnn) polynucleotide sequence selected from the group consisting of:vanin-1 (vnn1), wherein said vnn1 polynucleotide sequence has at least 70% sequence coverage to SEQ 3 or SEQ 98, and at least 70% sequence identity to SEQ 3 or SEQ 98;vanin-2 (vnn2), wherein said vnn2 polynucleotide sequence has at least 70% sequence coverage to SEQ 100, and at least 70% sequence identity to SEQ 100; andvanin-3 (vnn3), wherein said vnn3 polynucleotide sequence has at least 70% sequence coverage to SEQ 141, and at least 70% sequence identity to SEQ 141;ii. a cysteamine dioxygenase (ado) polynucleotide sequence which has at least 70% sequence coverage to SEQ 1, and at least 70% sequence identity to SEQ 1; andiii. a flavin-containing monooxygenase 1 (fmo1) polynucleotide sequence which has at least 70% sequence coverage to SEQ 5 or SEQ 99, and at least 70% of sequence identity to SEQ 5 or SEQ 99.

Described herein is a method of determining affinity of an antigen binding protein binding to a target, the method comprising: a) expressing an antigen binding protein in a prokaryotic host cell in a multi-well container, b) lysing the prokaryotic cell to produce a prokaryotic cell lysate; c) affixing the prokaryotic cell lysate to a substrate; and d) determining the affinity of the antigen binding protein to the target.

The invention discloses a recombinant cubilose peptide and application thereof, and belongs to the technical field of recombinant peptide preparation. The amino acid sequence of the recombinant cubilose peptide is as shown in SEQ ID NO.1, and the nucleotide sequence for coding the recombinant cubilose peptide is as shown in SEQ ID NO.2. The preparation method of the recombinant cubilose peptide comprises the following steps: (1) cloning a nucleotide sequence for coding the recombinant cubilose peptide into a prokaryotic or eukaryotic expression vector to construct a recombinant expression plasmid; (2) transforming or transfecting the recombinant expression plasmid into a proper expression host cell, wherein the expression host cell is a prokaryotic cell, a eukaryotic cell or a yeast cell; (3) culturing the expression host cell, and performing induced expression or expression under proper conditions; (4) collecting an expression product, crushing or cracking, and centrifuging to obtain a supernatant or precipitate containing the recombinant cubilose peptide; and (5) purifying the recombinant cubilose peptide. The cubilose peptide is prepared by using a recombinant expression technology, the cubilose peptide has remarkable anti-aging, tightening and moisturizing effects, and a foundation is laid for industrial production of the cubilose peptide.

The invention relates to methods for the flexible regulation of cellular growth of eukaryotic and prokaryotic cells. In particular embodiments, regulation of eucaryotic and microbial cells growth, occurs by the control of their interaction with the environmental factors, nutrient media, media additives, supplements.

Provided herein are nucleic acid constructs that comprise an inducible promoter. Dual expression systems are provided comprising two nucleic acid constructs or a single nucleic acid construct with two inducible promoters. Also provided are methods of expressing transcripts by transforming a nucleic acid construct described herein into a prokaryotic cell and contacting the prokaryotic cell with an inducer. Methods of producing a carotenoid are also disclosed herein.

The invention relates to methods for the flexible regulation of cellular growth of eukaryotic and prokaryotic cells. In particular embodiments, regulation of eucaryotic and microbial cells growth, occurs by the control of their interaction with the environmental factors, nutrient media, media additives, supplements.

The present application relates to the field of nucleic acid detection technology, and in particular to an integrated consumable device and method for integrating nucleic acid sample preparation and volume measurement. First, a sample processing tube containing a liquid sample is docked to a mounting ring, and then a heater is used to heat the liquid sample in the sample processing tube, and a sufficiently high temperature is generated in the sample processing tube to lyse eukaryotic cells or prokaryotic cells or viruses in the sample, and release nucleic acids. The temperature can also be high enough to denature proteins and inactivate pathogens. The elevated temperature can generate sufficient internal gas pressure, which is sufficient to pass the liquid sample through the connecting groove and the three-way valve into the metering container. The liquid sample can fill the space of the metering container, and the liquid will contact the actuator to activate the actuator, causing the three-way valve to shift to close the liquid outflow of the sample processing tube, and at the same time open the channel between the metering container and the reaction tube, so that the metered sample is moved into the reaction tube connected to the docking ring.

The invention relates to a method, expression system and related kit for producing at least one biologically active molecule, in particular a biotherapeutic, by optogenetic control of gene expression in at least one eukaryotic or prokaryotic cell, wherein the biologically active molecule comprises at least one protein of interest comprising at least two different subunits. In particular, one embodiment of the expression system according to the invention comprises three expression cassettes (2, 3, 4) for optogenetic control of the production of a protein of interest comprising two subunits (5, 6). A first expression cassette (2) comprises a first nucleic acid sequence (7) that is operably linked to a first promoter (8) and encodes a photosensitive transcription factor (9) comprising a gene expression controlling domain (10). The expression system (1) further comprises a second expression cassette (3) comprising a second nucleic acid sequence (11) encoding the first subunit (5) of the protein of interest. The second nucleic acid sequence (11) is operably linked to a second promoter (12) being operably linked to a transcription factor binding site (13) designed to bind the photosensitive transcription factor (9). The expression system (1) also comprises a third expression cassette (4) comprising a third nucleic acid sequence (15) encoding the second subunit (6) of the protein of interest and being operably linked to a third promoter (16).

Disclosed herein are polynucleotides comprising sequences encoding coenzyme M synthase (ComF) linked to a heterologous regulatory element and methods of using the same. The polynucleotides may comprise synthetic operons comprising additional sequences encoding enzymes, e.g., a taurine-pyruvate aminotransferase, a sulfoacetaldehyde acetyl transferase, or a sulfopyruvate decarboxylase. Also disclosed herein are recombinant prokaryotic cells, e.g., recombinant bacterial, e.g., E. coli, or archaeal cells, e.g., Methanosarcina acetivorans with improved tolerance to oxidative stress.

An engineered DNA molecule capable of being replicated in a cell, comprising a poly (A) tail coding sequence that makes the engineered DNA molecule more conservative when replicated in cells, particularly in prokaryotic cells, while adjusting the expression level of RNA in eukaryotic cells. Also provided are an RNA comprising the poly (A) tail and a use thereof.

The invention provides an expression and secretion system, and methods of using the same, for the expression and secretion of one fusion protein in prokaryotic cells and a second fusion protein in eukaryotic cells. Also provided herein are nucleic acid molecules, vectors and host cells comprising such vectors and nucleic acid molecules.

The present invention discloses a novel genome editing tool EbCas12a-D141R and its application in gene editing, belonging to the field of biotechnology. The EbCas12a-D141R is a variant of the type V class II CRISPR protein EbCas12a, and its amino acid sequence is as shown in SED ID NO.1 in prokaryotic cells and as shown in SED ID NO.2 in eukaryotic cells. The present invention for the first time obtains a variant EbCas12a-D141R with higher efficiency based on the type V class II CRISPR protein EbCas12a (1158AA), and the gene editing efficiency of this variant in eukaryotic cells is significantly higher than that of EbCas12a. The discovery of EbCas12a-D141R further expands the types of gene editing tools and has a very important role in basic scientific research and clinical treatment.

The application relates to a biota sesquiterpene synthase PoTPS1 and a coding gene thereof, the full length of the gene is 1746 bp, the gene codes 582 amino acids, the molecular weight of the enzyme is 67.9 kDa, a pSP-GM1-URA yeast protein expression plasmid is used as a carrier, saccharomyces cerevisiae CEN.PK-5D-113 is used as a host, the biota sesquiterpene synthase PoTPS1 is realized in the heterologous expression of eukaryotic cells, and two kinds of beta-himachalene and cedrol sesquiterpene alcohols can be generated at the same time; a pET22b escherichia coli protein expression plasmid is used as a carrier, anti-freezing protein XXA is added as a fusion tag, escherichia coli BL21 (DE3) is used as a host, the biota sesquiterpene synthase PoTPS1 is realized in the heterologous expression of prokaryotic cells, and a sesquiterpene compound is generated by taking ammonium salt FPP as a substrate.

The invention provides a genetic engineering modification method for improving acetic acid generation and application of the genetic engineering modification method. Specifically, provided herein is a recombinant prokaryotic cell that is genetically engineered directed to comprise a defect of one or more genes selected from the group consisting of galK, galT, galE, modF, modE, acrZ, modA, modB, modC, ybhA, and pgl wherein the recombinant prokaryotic cell has reduced acetic acid production as compared to the prokaryotic cell before modification. Also provided herein is a method of making the recombinant prokaryotic cell comprising directed genetic engineering of the prokaryotic cell to deficit one or more genes therein selected from the group consisting of galK, galT, galE, modF, modE, acrZ, modA, modB, modC, ybhA and pgl wherein the recombinant prokaryotic cell has reduced acetic acid production compared to the prokaryotic cell before modification. The invention also provides a product containing the recombinant prokaryotic cell, an application of the product in metabolic engineering and a method for carrying out recombinant production by utilizing the recombinant prokaryotic cell.