56 results about "Prokaryotic cells" patented technology

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 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.

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.

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.

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 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.

The present invention discloses a rabbit polyclonal antibody against Chlamydomonas reinhardtii NPHP4 protein, its preparation method and application. The amino acid sequence of the NPHP4 protein is shown in SEQ ID NO.1. Preparation method: Analysis and design of the antigen epitope of Chlamydomonas reinhardtii NPHP4 protein, and selection of the 859-1058aa amino acid fragment to construct a prokaryotic cell recombinant expression vector; transferring the recombinant expression vector into prokaryotic expression cells to obtain a recombinant expression strain and inducing the expression of the antigen protein; using the purified antigen protein as an immunogen to immunize rabbits to obtain a rabbit polyclonal antiserum against the NPHP4 protein, and further purifying it; the purified polyclonal antibody can be used to verify the labeling of the ciliary transition zone of Chlamydomonas reinhardtii by the IF method, proving that it can be used for specific fluorescence labeling of the ciliary transition zone; it can also be used for qualitative and quantitative analysis of the Chlamydomonas reinhardtii NPHP4 protein by the immunoblotting WB method.

The invention discloses a method for specifically separating eukaryotic cells from prokaryotic cells in various environments. The invention provides a concanavalin A and magnetic bead complex. The concanavalin A and magnetic bead complex is a complex formed by connecting concanavalin A and magnetic beads, the invention also provides a method for separating eukaryotic cells and prokaryotic cells, which comprises the following steps: adding the concanavalin A and magnetic bead complex into a cell resuspension to be separated, and incubating; and separating the eukaryote cells adsorbed by the magnetic beads from the prokaryote cells not adsorbed by the magnetic beads by using a magnetic frame. The raw materials such as the streptavidin magnetic beads and the concanavalin A involved in the method are low in price and simple to prepare; magnetic separation is utilized, and operation is simple and fast; the separation effect is good.

Disclosed herein is a nucleic acid delivery vector comprising capsid proteins or portions thereof from a non-enveloped mammalian virus, wherein the capsid proteins or portions thereof encapsidate a nucleic acid payload and wherein the capsid proteins or portions thereof were expressed and functionally processed in prokaryotic cells. Also disclosed is a method of producing a nucleic acid delivery vector. The method comprises: expressing capsid proteins or portions thereof from a non-enveloped mammalian virus in prokaryotic cells; optionally purifying the expressed capsid proteins or portions thereof from the prokaryotic cells; combining the expressed capsid proteins or portions thereof with a nucleic acid comprising: a nucleic acid payload; and a DNA packaging sequence of the non-enveloped mammalian virus, wherein the capsid proteins or portions thereof self-assemble and encapsidate the nucleic acid payload forming the nucleic acid delivery vector.

The invention discloses a dual expression vector, and a construction method and application thereof. The double expression vector comprises a transcription and translation element, a replication element and the like; wherein the transcription and translation element comprises an enhancer, an eukaryotic promoter, a prokaryotic promoter, a ribosome binding site, a spacer region, an initiation codon, a cloning site, a termination codon, an eukaryotic transcription termination sequence, a prokaryotic transcription termination sequence and the like which are sequentially connected from upstream to downstream. The same target protein can be efficiently expressed in a pronucleus-based cell-free protein expression system and a eukaryotic system by utilizing the double expression vector, the operation is simple and convenient, the efficiency is high, and the cost is low.

The invention provides a method for introducing a nanoparticle (100) into a prokaryotic cell (10), wherein the method comprises exposing the prokaryotic cell (10) to the nanoparticle (100), wherein: the nanoparticle (100) has an equivalent spherical diameter selected from the range of -20 nm; the nanoparticle (100) comprises a single-chain polymer nanoparticle (101); the nanoparticle (100) comprises a backbone (110), a plurality of intramolecular crosslinks (120), and a plurality of side groups (130) bound to the backbone (110); wherein the backbone (110) comprises n backbone atoms, wherein the n backbone atoms comprise ≥ 0.9*n carbon atoms; each intramolecular crosslink (120) covalently interconnects two backbone locations (112) of the backbone (110); k of the plurality of side groups (130) comprise ligand moieties (140) selected from the group comprising sugar moieties, a glycerol moiety, an acetate moiety, a pyruvate moiety, a lactate moiety, a succinate moiety, amino acid moieties, and fatty acid moieties, wherein k / n is selected from the range of 0.05-0.49; the prokaryotic cell (10) comprises a receptor (14) configured to interact with one or more of the ligand moieties (140).

A live cell-laden microgel, in accordance with one aspect of the present invention, includes at least one live cell and a photocured resin. An in-air drop formed microgel, in accordance with one aspect of the present invention, includes a component selected from the group consisting of DNA, RNA, a protein, a peptide, an antibody, a living-cell produced catalyst, a nucleic acid-based material, a prokaryotic cell, an eukaryotic cell, and a different type of biological component.

A method, an electrochemical sensor and a system for selective detection of infections. The method detects a concentration of nicotinamide adenine dinucleotide, NADH, from a bacterial culture through a cyclic voltammetry or chronoamperometry applied to an electrochemically active polymer. Therefore, prokaryotic cells can be detected while eukaryotic cells remain undetected. The infections can include bacterial infections and fungi or yeasts microbial infections.

Disclosed are methods, systems, components, and compositions for cell-free synthesis of glycosylated proteins. The glycosylated proteins may be utilized in vaccines, including anti-bacterial vaccines. The glycosylated 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 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 method of producing an organosulfur compound from a prokaryotic cell wherein said method comprises the steps of:a. providing a live prokaryotic cell capable of expressing at least one gene for the production of said organosulfur compound;b. exposing said live prokaryotic cell to a culture media with a pH of between 4 and 11 containing a carbon source and a sulfur source thereby creating an incubation mixture;c. incubating said live prokaryotic cell in said incubation mixture under aerobic or anaerobic conditions at a temperature ranging from 0° C. to 60° C. for a period of time sufficient for the expression of said at least one gene for the production of said organosulfur compound;d. recovering said organosulfur compound from the bacterial cells and / or spent media; ande. optionally, re-exposing said live prokaryotic cell to an unused media or spent media for the continuous production of said organosulfur compound of interest.

Provided is a self-selection DNA editing system. A subject self-selection DNA editing system includes: (a) a gene editing tool, and (b) a donor DNA comprising a bacteriocin unit, which includes a nucleotide sequence encoding a bacteriocin protein and / or a nucleotide sequence encoding a bacteriocin immunity protein. In some cases, (a) and (b) are present on the same nucleic acid (e.g., plasmid DNA). When a bacteriocin unit of the subject disclosure is present on (as part of) a donor DNA, the bacteriocin unit is configured for integration into a target DNA such as a genomic DNA. The present disclosure also provides methods of modifying a microbial community and methods of editing a prokaryotic cell to facilitate selection. The targeted prokaryotic cell(s) can be in culture or can be in situ (e.g., part of a microbial community).

The present invention relates to a cryopreservation formulation comprising collagen hydrolysate and preferably comprising dimethylsulfoxide. The present furthermore relates to a method for cryopreservation of one or more biological materials, the method comprising the step of providing the one or more biological materials in the cryopreservation formulation comprising collagen hydrolysate and preferably dimethylsulfoxide. The present invention also relates to a use of the cryopreservation formulation comprising collagen hydrolysate and preferably dimethylsulfoxide for cryopreservation of one or more biological materials selected from the group consisting of an eukaryotic cell, a prokaryotic cell, a cell organelle, an extracellular vesicle, an organoid, a tissue, and an organ. The present invention also relates to a use of collagen hydrolysate, preferably in combination with dimethylsulfoxide, in preparing a cryopreservation formulation.

The present invention addresses the problem of, in a method for extracting a target protein from prokaryotic cells expressing the target protein, preventing the contamination of the target protein with foreign substances and increasing the recovery rate of the target protein.　The method for extracting a target protein from prokaryotic cells that is disclosed in the present description is characterized by comprising: a step for mixing a culture solution containing prokaryotic cells that express the target protein with a carboxylic acid at a final concentration of 0.5-15% (v / v) inclusive to prepare a mixed liquid; and a step for extracting the target protein from the prokaryotic cells in the mixed liquid.