26 results about "Prokaryote" patented technology

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.

Novel combinations of cysteine-rich insecticidal proteins (CRIPs) and insecticides (IAs), such as chemicals, molecules, nucleotides, polynucleotides, peptides, polypeptides, proteins, toxins, toxic substances, poisons, insecticides, pesticides, organic compounds, inorganic compounds, prokaryotes or eukaryotes (and substances produced from such prokaryotes or eukaryotes), are described and claimed for the control and / or eradication of pests. [Solution] Novel insecticide combinations, compositions, and methods for using them are provided. The present invention relates to a combination of a cysteine-rich insecticide peptide (CRIP) and an insecticide (IA). The present invention also describes the use of the combination and composition for controlling insects. This specification describes the gene encoding CRIP, compositions and combinations containing CRIP and IA, and methods useful for controlling pests using them.

The invention provides an anti-tumor peptide modified bacterial outer membrane vesicle and a preparation method thereof, and relates to the technical field of engineering bacterial outer membrane vesicles. According to the invention, a recombinant plasmid capable of expressing a ClyA-A25 fusion protein in a prokaryote is designed and constructed, and the engineering bacterium outer membrane vesicle W-A25OMV modified by the anti-tumor peptide A25 is successfully prepared by using the recombinant plasmid. The W-A25OMV retains the basic structure of bacterial outer membrane vesicles, enhances the anti-tumor biological activity of A25 peptide, has good stability, shows a remarkable inhibition effect on invasion and migration of tumor cells, can effectively promote apoptosis of the tumor cells, and has development and application potential as an anti-tumor drug.

The application discloses a novel lactoferrin inducible soluble expression system and a construction method thereof, and belongs to the technical field of biotechnology and genetic engineering, and the specific scheme is that human lactoferrin (LF) is expressed by probiotic Escherichia coli Nissle1917, the protein is purified after the expression position is determined, and anti-inflammatory activity is determined. The LF expression system constructed by the application provides a construction method of a prokaryotic expression stable human lactoferrin expression vector, the vector is introduced into probiotic Escherichia coli Nissle1917, soluble target recombinant human lactoferrin is obtained after induction fermentation expression, two-step purification and identification, and the application has the characteristics of high safety, low cost, easy operation and the like.

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 belongs to the technical field of bioengineering, and particularly relates to a preparation method and application of an N protein polyclonal antibody of a horse coronavirus. The polyclonal antibody is prepared by truncation expression of the horse coronavirus N protein, and a material is provided for research on inhibition of host innate immune mechanism by the horse coronavirus N protein. Prokaryotic expression is carried out on the N protein of the equine coronavirus, the polyclonal antibody is prepared, and a method and a material are provided for molecular research and mechanism research of the equine coronavirus. According to the invention, in combination with bioinformatics analysis of an N protein of the horse coronavirus, a pGEX-6p-N3 recombinant plasmid is constructed, and prokaryotic expression is carried out, so that a purified recombinant protein is obtained. After a mouse is immunized by the recombinant protein, a polyclonal antibody is obtained, an indirect ELISA method detects that the polyclonal antibody has good titer, Western Blot detection has good reactivity, and the polyclonal antibody can be applied to an indirect immunofluorescence test.

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.

The invention relates to a modular culture system for growing prokaryotic and / or eukaryotic organisms using a nutrient solution (N) for the organisms, wherein the culture system comprises a series of culture modules (100) for receiving the organisms, which culture modules are interconnected to conduct the nutrient solution (N), and an extraction module (200), connected to one of the culture modules (100) for conducting the nutrient solution (N), for the controlled extraction of the nutrient solution (N) from the culture module (100).Each culture module (100) comprises: an outer container (110) through which the nutrient solution (N) flows in a flow direction (S); an inner container (120), located at least partially within the outer container (110), for receiving the organisms; and several connection devices (130) for connecting the outer container (110) to the outer container (110) of an additional culture module (100) and / or to the extraction module (200) for conveying the nutrient solution (N). The inner container (120) is permeable to the nutrient solution (N), and a nutrient chamber (115), through which the nutrient solution (N) flows, is located between the outer container (110) and the inner container (120). The extraction module (200) comprises an extraction control device (240) for controlling the flow of the nutrient solution (N) extracted from the cultivation module (100) by means of the extraction module (200).

The application belongs to the field of animal bacteriology and molecular biology technology, and relates to preparation of a hybridoma cell and application thereof. A monoclonal antibody is prepared through intracellular Lawsonia intracellularis outer membrane protein OmpA expressed by prokaryotes, and IFA, WB and blocking ELISA and other immunological detection methods for detecting or diagnosing Lawsonia intracellularis are established through the expressed intracellular Lawsonia intracellularis outer membrane protein OmpA and the prepared monoclonal antibody. The blocking ELISA for detecting serum antibodies of Lawsonia intracellularis can detect serum samples of suspected Lawsonia intracellularis infection from different species, and does not exist cross reaction with positive serum of common pig pathogens, has high sensitivity and specificity, is good in repeatability, and is suitable for high-throughput detection of serum samples.

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.

The invention discloses preparation and application of oligopeptide immune carrier protein, and belongs to the technical field of biology. The method comprises the following steps: fusing a gene sequence for coding oligopeptide and a gene sequence for coding ferritin through a chemical method by utilizing the characteristics that oligopeptide molecules are small in mass and ferritin is self-assembled to form a polymer, and then cloning the fused gene into a prokaryotic biological carrier for expression; the recombinant protein is purified by using an expression system which is not limited to prokaryotes such as escherichia coli and eukaryotes. The protein has the advantages of low preparation cost, strong immunogenicity and the like, and lays a foundation for obtaining a high-affinity anti-oligopeptide nano antibody.

According to the eukaryote level gene transfer identification method and device, the storage medium and the computer equipment provided by the invention, after the target protein sequence of the high-quality genome of the target eukaryote is extracted, the target protein sequence can be compared with the protein sequence in the NR database; screening the protein sequences in the comparison result according to a preset screening condition to obtain a plurality of candidate sequences corresponding to the target protein sequence and a donor sequence corresponding to each candidate sequence, and then performing multi-sequence comparison on each candidate sequence and the corresponding donor sequence, and constructing a corresponding gene topological structure according to a multi-sequence comparison result of each candidate sequence, and determining a biological group in which each gene topological structure is located, so that the candidate sequence, in which the gene topological structure is located in the prokaryotic biological group, in each candidate sequence can be used as a gene sequence from prokaryotic horizontal transfer. And according to a gene topological structure, automatically eliminating non-conforming candidate sequences.

The application provides a genetically engineered bacterium for producing recovery promoting factor Rpf and application thereof, relates to the technical field of bioengineering, and belongs to prokaryotes. The nucleotide sequence of the genetically engineered bacterium is shown in SEQ ID NO. 1. The sequence is obtained by codon mutation and DNA sequence optimization of a wild-type bacterium recovery promoting factor of Micrococcus luteus in an original nucleotide sequence, and the optimized nucleotide sequence is named Delta Rpf. The original nucleotide sequence is shown in SEQ ID NO. 2. The purity of the Delta Rpf recombinant protein of the application can reach 93 %, which is much higher than the purity of the protein extracted by the prior art. The enzymatic activity of the Delta Rpf recombinant protein prepared by the genetically engineered bacterium constructed by the application is significantly higher than the enzymatic activity of the unmutated Rpf protein.

The invention provides a construction method of an expression vector of eukaryotic genes in prokaryotes, and belongs to the field of molecular biology. The method comprises the following steps: (a) designing a PCR (Polymerase Chain Reaction) primer pair for amplifying each exon in a specific transcription spliceosome cds sequence; (b) respectively amplifying each exon by using the PCR primer pair designed in the step (a) to obtain a PCR fragment of each exon; (c) mixing the n exon fragments obtained in the step (b) as a template, and carrying out overlapping PCR by using the forward primer P1-F of the first exon and the reverse primer Pn-R of the final exon to obtain a complete cds fragment of the specific transcription spliceosome; and (d) purifying and recovering the cds fragment of the specific transcription spliceosome obtained in the step (c), and connecting the cds fragment to a prokaryotic expression vector. Therefore, the problem that the cds fragment of the target transcription spliceosome cannot be obtained due to unknown temporal-spatial expression information of a luxury gene or a specific transcript is solved.

The application discloses a metal ion-dependent PET depolymerase Sv10 mutant and application thereof, and relates to a PET depolymerase 10 derived from a prokaryotic variant Saccharophagus degradans Saccharothrix variisporea Sv 10, the amino acid sequence of which is shown in SEQ ID NO. 2, is subjected to mutation to obtain a mutant with significantly improved activity, thermal stability and degradation effect compared with the original enzyme 10. Sv The application also relates to application of the PET depolymerase 10 and the mutant thereof in degradation of broken waste polyester. Sv Sv Sv Sv Sv Sv Sv S The PET depolymerase and the mutant thereof disclosed by the application show significant metal ion dependence, and the Ca 2+ can obviously activate the depolymerization activity, and has a wide application prospect in a metal-containing reaction system.​

The invention relates to the field of in-vitro transcription, in particular to a 5 '-UTR element, an expression vector and application of the 5'-UTR element. The invention provides a 5 '-UTR element. The 5'-UTR element has a nucleotide sequence as shown in SEQ ID NO: 5, a nucleotide sequence as shown in SEQ ID NO: 6 and a nucleotide sequence as shown in SEQ ID NO: 7. Or a nucleotide sequence which is obtained by substituting, deleting or adding one or more basic groups to the nucleotide sequence as shown in the specification, and has the same or similar function as the nucleotide sequence as shown in the specification; or a nucleotide sequence having at least 80% of identity with the nucleotide sequence shown in the description. By providing the 5 '-UTR sequence, the problem that when mRNA is synthesized in vitro, the 5'-UTR sequence of a prokaryote cannot have the functions of high mRNA expression efficiency and high capping efficiency at the same time is solved.

This invention provides an engineered Cas13 nuclease, a gene expression regulation system, and its applications. The engineered Cas13 nuclease includes engineered Cad13d or Cas13a nucleases. The engineered Cas13d nuclease is obtained by truncating the amino acid sequence of the RfxCas13d nuclease by any one of the following: 249-255, 250-255, 251-255, or 252-255. The engineered Cas13a nuclease is obtained by truncating the amino acid sequence of the LwaCas13a nuclease by any one of the following: 485-491, 486-491, 1057-1061, or 1058-1061. It also provides a gene expression regulation system for achieving high-fidelity RNA-level gene knockdown in vivo and a method for RNA-level gene knockdown applicable to prokaryotes. The engineered Cas13 nuclease of this invention can achieve higher fidelity cleavage.

The present application relates to the field of biotechnology, in particular to a preparation method of sound-sensitive stem cell-derived beta functional cells and application thereof. The preparation method provided by the present application introduces a prokaryote-derived engineered mechanosensitive ion channel MscL-G22S into human pluripotent stem cells, and combines with a directional differentiation technology to obtain pancreatic islet beta-like cells with ultrasonic response function. The cell product not only has a natural glucose-dependent insulin secretion capacity, but also has an additional function of being activated by low-intensity focused ultrasound in a non-invasive manner through the introduction of an exogenous ion channel. This double regulation mechanism significantly improves the functional maturity and response sensitivity of the transplanted cells in vivo, and can achieve a better blood glucose control effect under the same number of cells, thereby providing a technical possibility for reducing the amount of transplanted cells, reducing the volume of transplanted cells and expanding the transplanted site, and being suitable for popularization and application.

The application belongs to the technical field of biology and relates to a nanobody or antigen-binding fragment thereof specifically binding to pig IgM Fc receptor FcμR and application. The application clones full-length cDNA of pig FcμR, expresses the extracellular region in prokaryotes, and constructs a nanobody library by camel immunization; a strain of specific anti-pig FcμR nanobody is obtained through three rounds of panning. The nanobody (FcμR-VHH) is expressed by E. coli, and it is proved that the nanobody can target pig macrophages (PAMs) by taking zsGreen as a model. In vitro and in vivo tests show that the targeting and immune enhancement effect of the FcμR-VHH fusion protein at the C-terminal of an antigen is better than that at the N-terminal. After immunizing animals, ELISA proves that the fusion protein significantly improves humoral and cellular immune responses.

The present application belongs to the technical field of protein purification. Provided in the present application are a renaturation-assisting tag for purifying an inclusion body protein, and a gene-derived product thereof and the use thereof. Provided in the present application is a renaturation-assisting tag P67, comprising at least one of the amino acid sequences as set forth in SEQ ID NO: 1 to SEQ ID NO: 3 and mutants thereof, wherein the mutant has a mutation at position 33 of the amino acid sequence from lysine to glutamic acid or proline. The renaturation-assisting tag P67 according to the present application is fused and expressed with a target protein in a prokaryote, enabling successful renaturation of a recombinantly expressed inclusion body (insoluble inclusion body) and conferring a biological activity thereon. The renaturation-assisting tag P67 establishes a foundation for subsequent protein functional research and a translational use.

The invention relates to an AnaCas9 mutant as well as a gene editing system and application thereof, and relates to the technical field of biology. The AnaCas9 mutant is modified at a specific site, so that the gene editing activity of the AnaCas9 mutant is remarkably improved, and the AnaCas9 mutant can be widely applied to efficient gene editing of prokaryotes and eukaryotes and particularly can also realize efficient gene editing in mammalian cells (including human cells) and saccharomycetes.

The invention discloses application of selenoprotein O in lipid metabolism regulation and mitochondrial protection, and belongs to the technical field of biochemistry. The technical problem to be solved is that specific mechanisms of lipid metabolism regulation and mitochondrial stress protection in the prior art are not clear. According to the key point of the technical scheme, the application of selenoprotein O in lipid metabolism regulation and mitochondrial protection is provided, NAD < + > in mitochondria is hydrolyzed into NMN and AMP under the action of SelO, Mn < 2 + > is used as a cofactor, the reaction is sensitive to enhanced mitochondrial matrix pH reaction, the mitochondria is protected from continuous excessive metabolism activation, and in addition, the selenoprotein O can be used as a lipid metabolism regulator. The SelO directly interacts with fatty acid oxidation related enzymes to inhibit lipid catabolism. The discovery of the application reveals a conservative space-time NAD + regulation mechanism, and highlights the physiological importance of the space-time NAD + regulation mechanism in prokaryotes and eukaryotes; the kit or the medicine for adjusting the activity of the selenoprotein O in the mitochondria can be used for preparing products for preventing or treating mitochondria-related diseases or preparing weight-losing products.

This invention discloses the CsGSTU8 gene of tea plant and its application in alleviating and degrading glyphosate toxicity, relating to the field of genetic engineering technology. The nucleotide sequence of the CsGSTU8 gene is shown in SEQ ID NO.1 of the sequence listing. The amino acid sequence of the protein encoded by the CsGSTU8 gene is shown in SEQ ID NO.2 of the sequence listing. CsGSTU8 is upregulated in both roots and leaves in response to glyphosate stress; CsGSTU8 is specifically highly expressed in tea plant roots. The pCAMBIA1305 plasmid (containing a GFP tag) constructed from this gene, after transformation with Agrobacterium and infection of tobacco leaves, was observed to be localized in the nucleus and cytoplasm. The CsGSTU8 protein is expressed in prokaryotes, and its enzyme activity is strongest at pH 8.0. Transforming the pYES2 plasmid constructed from this gene into the glyphosate-sensitive strain BY4741 restored the growth ability of the yeast mutant on high-concentration glyphosate medium. The cloning of this gene elucidates its role in tea plant resistance to glyphosate stress, providing data support for research on the molecular mechanisms of tea plant resistance to glyphosate stress.

The invention provides a genetically engineered bacterium for producing a resuscitation promoting factor Rpf and application of the genetically engineered bacterium, and relates to the technical field of bioengineering, the genetically engineered bacterium belongs to prokaryotes, the nucleotide sequence of the genetically engineered bacterium is as shown in SEQ ID NO.1, and the nucleotide sequence of the genetically engineered bacterium is as shown in SEQ ID NO.2. The sequence is obtained by carrying out codon mutation and DNA (Deoxyribose Nucleic Acid) sequence optimization on an original nucleotide sequence of a micrococcus luteus wild type bacterium resuscitation promoting factor, and the optimized nucleotide sequence is named as Delta Rpf; 1, and the original nucleotide sequence is as shown in SEQ ID NO. 2. The purity of the Delta Rpf recombinant protein can reach 93% and is far higher than that of protein extracted in the prior art. The activity of Delta Rpf recombinant protease prepared from the genetically engineered bacterium constructed by the invention is obviously higher than that of unmutated Rpf protease.

The invention relates to the field of in-vitro transcription, in particular to a 5 '-UTR element, an expression vector and application of the 5'-UTR element. The invention provides a 5 '-UTR element. The 5'-UTR element has a nucleotide sequence as shown in SEQ ID NO: 4, a nucleotide sequence as shown in SEQ ID NO: 5 and a nucleotide sequence as shown in SEQ ID NO: 6. Or a nucleotide sequence which is obtained by substituting, deleting or adding one or more basic groups to the nucleotide sequence as shown in the specification, and has the same or similar function as the nucleotide sequence as shown in the specification; or a nucleotide sequence having at least 80% of identity with the nucleotide sequence shown in the description. By providing the 5 '-UTR sequence, the problem that when mRNA is synthesized in vitro, the 5'-UTR sequence of a prokaryote cannot have the functions of high mRNA expression efficiency and high capping efficiency at the same time is solved.

A technique is provided that uses protoporphyrinogen oxidase or its amino acid variants derived from prokaryotes to confer enhanced herbicide tolerance and / or improve herbicide tolerance in plants and / or algae.