115 results about "Exogenous protein" patented technology

This invention provides an in vitro cell-free protein synthesis system, its reagent kit, and its applications, belonging to the field of protein synthesis technology. This invention improves an in vitro cell-free protein synthesis system based on Kluyveromyces lactis by adding exogenous L-arabinose, significantly enhancing the system's protein synthesis capacity. Furthermore, it provides a more efficient and higher-throughput in vitro protein synthesis reagent kit and a method for synthesizing exogenous proteins. The improvements provided by this invention can be achieved without molecular modification, are simple and convenient to operate, and save costs.

The invention belongs to the technical field of genes, and discloses a method for efficiently expressing a foreign protein based on an NC048604-1 site in a CHO cell genome. A site for stably expressing protein in the CHO cell genome is located in the 94142000 to 94148000 basic group range of the CHO cell genome NC048604-1, and the nucleotide sequence of the site is as shown in SEQ ID NO: 1. According to the invention, different protein genes are introduced at fixed positions in a CHO cell genome, and stable expression is carried out.

The application provides an in-vitro cell-free protein synthesis system and kit containing exogenous magnesium ions and application thereof, including a D2P system (DNA-to-Protein system) and an mR2P system (mRNA-to-Protein system), and belongs to the technical field of protein synthesis. The in-vitro cell-free protein synthesis system containing exogenous magnesium ions adopts magnesium aspartate as a novel magnesium ion source, and especially in a eukaryotic cell-free system, compared with a traditional magnesium ion source, can significantly improve the protein synthesis efficiency and protein expression amount. A more efficient and higher-throughput in-vitro protein synthesis kit and a synthesis method of exogenous proteins are also provided, and the kit has the advantages of simplicity, convenience and low cost.

The invention discloses a novel plasmid vector capable of increasing CHO foreign protein expression quantity and construction and application thereof, and belongs to the technical field of gene engineering. The plasmid vector pCHO6GS belongs to a brand-new plasmid system, and the expression quantity of a common monoclonal antibody and recombinant protein (such as H5HA trimer) difficult to express in CHO can be increased by at least more than one time, so that the expression quantity of foreign protein is increased beyond expectation through the novel plasmid vector; and the method also plays an extremely important role in reducing cost and improving efficiency of various protein products taking CHO as a cell factory.

The invention discloses a method for quickly rescuing bovine coronavirus epidemic strains and application of the method. The bovine coronavirus SHZ isolate is subjected to efficient segmented cloning, the obtained segment and a Linker sequence are subjected to a cyclic polymerase extension reaction, and the obtained reaction product can be directly transfected to 293T cells for virus packaging without purification. Different from a traditional method, the method does not need to amplify full-length virus cDNA clone by means of bacteria and yeast, but directly obtains a sufficient amount of preliminary products through PCR, and avoids the problems of instability and low efficiency when partial sequences of a virus genome proliferate in a bacteria or yeast host. By adopting the reverse genetic system, the recombinant bovine coronavirus expressing the foreign protein is successfully rescued, a brand new technical platform is provided for dynamic visualization research of in-vivo and in-vitro replication of the virus, and an efficient and flexible tool is also provided for virology research, development of virus vector vaccines and screening of antiviral drugs.

The invention provides a preparation method of a recombinant cell, the recombinant cell and application of the recombinant cell, and relates to the technical field of biology. A target protein expressed by the recombinant cell contains one, two, three or four polypeptide chains; the preparation method comprises the following steps: integrating exogenous polynucleotides for coding a target protein into a genetic material of a starting cell in a site-specific manner, wherein each exogenous polynucleotide for site-specific integration contains coding genes of all polypeptide chains of the target protein; each exogenous polynucleotide contains at least four expression cassettes, and each polypeptide chain is encoded by at least one expression cassette. The preparation method is simple to operate, coding genes of all polypeptide chains can be integrated into genetic materials of starting cells at a time, and the expression quantity of foreign proteins of the recombinant cells obtained through the preparation method is high.

To provide a new method for efficiently purifying an exogenous protein expressed in a plant.SOLUTION: A method for purifying a foreign protein expressed in a plant, comprising the step of adding an acid to an extract of a tissue of a plant in which a foreign protein is expressed to degrade contaminating proteins derived from the plant.SELECTED DRAWING: None

The invention discloses a protein sequence analysis method, equipment, a medium and a product, and relates to the technical field of combination of biotechnology and artificial intelligence. The method comprises the steps that a first protein sequence is obtained, corresponding first embedding expression is constructed, and the first protein sequence is an exogenous protein sequence which is expressed in Escherichia coli or is to be expressed in Escherichia coli and is not from Escherichia coli; inputting the first embedded representation into a pre-trained proteotoxicity analysis model to obtain a prediction result aiming at whether the first protein sequence is toxic to escherichia coli or not; the model is a convolutional neural network model based on a channel attention mechanism and a space attention mechanism. According to the technical scheme provided by the embodiment of the invention, the problem that a method for analyzing the toxicity of the escherichia coli by aiming at the protein sequence cannot give consideration to both efficiency and accuracy is solved, and the toxicity of the endogenous or exogenous protein sequence expressed in the escherichia coli to the escherichia coli can be efficiently, stably and accurately predicted and analyzed.

According to the method for efficiently identifying adulteration of the dairy products through the endogenous nanometer characteristics, natural nanometer particles existing in endogenous sources in the dairy products are detected, multiple nanometer characteristic indexes are integrated to form discrimination parameters, an artificial learning-machine algorithm is utilized to construct a discrimination model, the discrimination model is optimized, the dairy products to be detected are discriminated, and the adulteration of the dairy products to be detected is identified. The dairy product adulteration can be efficiently identified on the premise that adulterants are not known. According to the method, various dairy products including bulk milk and rare camel milk can be identified, a judgment model needs few optimization parameters, the fitting performance is good, the equation fitting degree can reach 0.98 or above, and the prediction effect is good. Experiments prove that the method provided by the invention has a relatively high discrimination rate on common adulterants such as foreign protein, starch and water in dairy products, and the discrimination rate can reach 90% or above.

The invention discloses a PVX expression vector pWX for efficiently expressing foreign protein and application of the PVX expression vector pWX. In the construction process of optimizing potato X virus infectious clone, after a single base mutation NruI-gene sequence (CCGCGA) is inserted into an upstream regulation region of a PVX coat protein subgene promoter, a novel PVX expression vector capable of promoting foreign protein expression is obtained, and the novel PVX expression vector is named as pWX. Experiments prove that compared with a control vector, the accumulation amount of GFP (Green Fluorescent Protein) generated by the pWX vector through transient expression of agrobacterium in tobacco is obviously increased. The invention further applies a pWX vector to successfully express an important target protein HPV16L1 researched and developed by an HPV vaccine, and the expression quantity of the important target protein HPV16L1 in tobacco is 5.25 micrograms per gram of fresh heavy leaves. The invention provides a novel vector based on a PVX genome for high-efficiency expression of a plant source target protein, and the vector has a good application prospect in preparation of plant source vaccines.

The invention discloses gluten-free rice bread and a preparation method thereof, and relates to the technical field of rice bread preparation, and the gluten-free rice bread comprises rice flour, glutinous rice flour, water, white granulated sugar, milk powder, butter, yeast and edible salt. According to the gluten-free rice bread, any complex modifying agent such as hydrophilic colloid, foreign protein or enzyme preparation does not need to be added, and the quality of the gluten-free rice bread is improved under the extremely simple formula.

The present application relates to the field of microbial technology, and particularly relates to a gene editing method of propionibacterium acnes VB208 and application thereof. Propionibacterium acnes ) on February 20, 2023, and is preserved in the China General Microbiological Culture Collection Center, with a preservation number of CGMCC No.26576. The propionibacterium acnes screened by the present application has the characteristics of skin surface occupation, high conversion efficiency, gene modification potential and efficient expression of exogenous proteins, and can be used as an engineering strain. Furthermore, a genetic operation system suitable for the strain can be established by using the propionibacterium acnes, and the application prospect is wide.

The invention provides an in-vitro cell-free synthesis method and kit of foreign protein, and belongs to the technical field of protein synthesis. According to the in-vitro cell-free synthesis method of the foreign protein, a water-phase in-vitro cell-free protein synthesis system and a nucleic acid template for coding the foreign protein are provided, a hydrophobic interface in surface contact with the water-phase in-vitro cell-free protein synthesis system is provided, incubation reaction is carried out, the foreign protein is synthesized, and the in-vitro cell-free synthesis method of the foreign protein is obtained. The protein synthesis efficiency and the protein expression quantity of the cell-free system are improved. The invention further provides an in-vitro protein synthesis kit which is more efficient and higher in throughput. The invention provides a simple and convenient hydrophobic interface optimization method which is simple and convenient to operate and saves cost.

The invention relates to a combined additive, a CHO cell culture medium, a preparation method and a culture method. The combined additive comprises cyclohexyl alanine and 3-(2-naphthyl)-L-alanine. The CHO cell culture medium comprises amino acids, vitamins, trace elements, inorganic salts, lipids, carbohydrates and a combined additive, the preparation method is used for preparing the CHO cell culture medium. The culture method uses a CHO cell culture medium to culture CHO cells. The combined additive is obtained through structural domain virtual docking and hydrophobicity regulation and control screening, G1 / S conversion can be mildly delayed, cell apoptosis can be inhibited, and an expression window can be prolonged. The culture medium contains amino acids, vitamins, trace elements and the like, and preferably selects a functional additive to replace serum and foreign protein functions. According to the application, high proliferation of CHO cells and efficient target protein expression can be realized under the conditions of no serum and no foreign protein, meanwhile, the accumulation of lactic acid and ammonia is reduced, the cell caking phenomenon is reduced, and the application has remarkable technical advantages.

The present disclosure provides a nucleic acid molecule comprising a 3' type II intron fragment, a 5' type II intron fragment, and a protein coding region or non-coding region, which can obtain a circular RNA with biological activity. The present disclosure also provides a vector comprising the nucleic acid molecule, and applications thereof. The self-splicing system constructed and optimized by the present disclosure determines that other type II intron self-splicing systems can be used to produce circular RNA and express exogenous proteins in vivo and in vitro, and the system has high ring formation efficiency and protein expression amount.

Nanoparticulate pharmaceutical formulations and methods for co-delivery of two or more species of nucleic acids for simultaneous suppression and expression of target genes in a cell, are provided. The nanoparticles encapsulate two or more nucleic acid species. The first nucleic acid suppresses expression of a gene or product thereof, e.g., inhibitory nucleic acid, such as antisense, siRNA, miRNA, Dicer siRNA, piRNA, etc. The second nucleic acid increases expression of, or encodes, an endogenous or exogenous protein or polypeptide, e.g., an mRNA. The first and second nucleic acid species simultaneously target or affect the same or different cellular processes within a cell including communication, senescence, DNA repair, gene expression, metabolism, necrosis, and apoptosis.

The application discloses a CMV virus-like particle for producing a VLP recombinant vaccine and a preparation method thereof, the CMV virus-like particle is obtained by cloning a C-terminal G4SLPETG modified plant virus cucumber mosaic virus CMV capsid gene into a prokaryotic expression vector to obtain a recombinant expression vector, the recombinant expression vector is transfected into E. coli BL21 (DE3), and expression is obtained through the recombinant E. coli BL21 (DE3); and the amino acid sequence of the C-terminal G4SLPETG modified CMV is SEQ ID NO. 1. Test proves that the recombinant strain constructed by the application is stable to expression of an exogenous protein. The recombinant protein expressed by the application is used for preparing a virus-like particle for antigen coupling, the coupling efficiency is high, the antigen purity is high, the safety is good, the virus-like particle is not pathogenic to mice and other animals, and the safety evaluation is easy.

The invention discloses an inflammation regulation sequence of dairy cow mastitis and application thereof, and belongs to the technical field of biology, the inflammation regulation sequence is a regulation sequence responding to a gram-negative bacterium inflammation signal, the regulation sequence is a promoter sequence of a CXCL2 gene and is shown as SEQ ID NO: 1, and the regulation sequence is a promoter sequence of a CXCL2 gene and is shown as SEQ ID NO: 2. A regulatory sequence responding to a gram-positive bacterium inflammation signal, which is a promoter partial sequence of the NLRP3 gene and is as shown in SEQ ID NO: 2; the regulatory sequence responding to a gram-positive bacterium inflammation signal is a promoter reverse complementary sequence of the TLR2 gene and is shown as SEQ ID NO: 3; the endogenous inflammation regulation sequence provided by the invention is composed of genome elements of the dairy cow, and no exogenous coding gene is introduced, so that potential risks caused by exogenous proteins are avoided.

The present invention relates to compositions comprising yeast oleosomes and at least 30 wt% exogenous protein, based on the total solids content of the composition, to food products or beverages comprising such compositions, to the use of yeast oleosomes to improve the sensory properties of a composition comprising at least 30 wt% exogenous protein based on the total solids content of the composition and to a process for the preparation of a composition comprising at least 30 wt% exogenous protein based on the total solids content of the composition.

The invention relates to the technical field of bioengineering, and discloses a saccharomyces cerevisiae self-replication system and application thereof.The system comprises a recombinant plasmid, and the recombinant plasmid comprises upstream and downstream homologous arms of a saccharomyces cerevisiae delta site, a selection marker TRP1, an autonomous replication sequence ARS310, a G418 resistance gene and a resistance gene promoter; wherein the nucleotide sequence of the autonomous replication sequence ARS310 is as shown in SEQ ID NO. 3; the nucleotide sequence of the G418 resistance gene is as shown in SEQ ID NO. 4; the nucleotide sequence of the selection marker TRP1 is as shown in SEQ ID NO. 5. The invention provides a pressure-driven saccharomyces cerevisiae self-replicating system for high-efficiency expression of foreign protein (target gene), and the expression of the foreign target gene with high conversion efficiency, high stability and high copy number is realized by utilizing natural multiple copies of a delta site, the self-replicating capability of ARS and the screening pressure of a G418 resistance gene.

The application discloses a silk material with a cell proliferation function and application thereof, the functional material is expressed by a human type I collagen truncated fragment in a middle silk gland of a domestic silkworm, a sericin promoter is used as a tissue-specific promoter to drive specific expression of an exogenous gene in the middle silk gland, and finally the exogenous gene is combined with sericin protein and is secreted to outside of the body, the exogenous protein can be separated after a certain treatment of a cocoon, and the expression product can keep biological activity. The silk material is used to prepare a hydrogel, has a function of promoting wound healing, and can be applied to aspects of promoting skin regeneration and repair, bone graft substitutes, cartilage reconstruction, artificial skin and the like.

Provided are a protein for increasing the expression yield of an exogenous protein and the use thereof, wherein an ERV29 protein and a related protein, and a P180 protein and a related protein can be used as molecular chaperones to improve the expression yield of exogenous proteins such as brazzein, the ERV29 protein has an amino acid sequence as shown in SEQ ID No. 4, and the P180 protein has an amino acid sequence as shown in SEQ ID No. 6.

The application discloses a novel plasmid vector with improved CHO exogenous protein expression amount and construction and application thereof, and belongs to the technical field of genetic engineering. The plasmid vector pCHO6GS belongs to a novel plasmid system, and can at least improve the expression amount of general monoclonal antibodies and difficult-to-express recombinant proteins (such as H5HA trimer) in CHO by more than 1 times, that is, the unexpected improvement of the exogenous protein expression amount is realized by the novel plasmid vector, and the novel plasmid vector has an extremely important role in cost reduction and efficiency increase of various protein products with CHO as a cell factory.

The invention provides a cell genome site capable of being used for targeted integration of exogenous nucleic acid molecules. The site is located in a nucleic acid sequence corresponding to a cell genome NC048595.1: 454997734-455024715 of a Chinese hamster ovary CHO-K1. The cell genome sites can integrate foreign protein genes and perform stable expression. The invention also provides stable and high-yield targeted integrated cells, such as host cells derived from Chinese Hamster Ovary (CHO), methods for their production and uses thereof.

The present application relates to the field of agricultural biotechnology, and in particular to a Pichia pastoris engineering strain with improved expression and secretion capacity and overexpression of ALG14 and a method for improving the expression amount of exogenous proteins, and the main technical problem to be solved by the present application is to overcome the deficiencies of the prior art, and to provide a strategy for improving the expression of exogenous proteins by Pichia pastoris, and to improve the enzyme production capacity of Pichia pastoris by overexpression of the AGL14 gene of Pichia pastoris itself.

The invention discloses a litopenaeus vannamei egg membrane outer layer protein VMO1 and an application thereof in egg entering of a prawn foreign protein. The amino acid sequence of the egg membrane outer layer protein VMO1 of the litopenaeus vannamei is as shown in SEQ ID NO. 2. The litopenaeus vannamei vitelline membrane outer layer protein gene LvVMO1 is cloned from litopenaeus vannamei for the first time, and experiments prove that the litopenaeus vannamei vitelline membrane outer layer protein gene LvVMO1 is generated in the hepatopancreas tissue of the litopenaeus vannamei and enters eggs through a hemolymph way. A VMO1-EGFP in-vivo tracing test further shows that VMO1 is generated in hepatopancreas of the litopenaeus vannamei, enters the ovary through hemolymph, can carry foreign proteins such as EGFP, is transferred from an abdominal injection site and is enriched in the ovary. The invention provides an effective technical means for introducing a gene editing system into litopenaeus vannamei egg cells by using foreign proteins to realize genetic improvement and other applications of litopenaeus vannamei, and has wide application prospects and great economic values.

Provided in the present disclosure is a nucleic acid molecule. The nucleic acid molecule contains a 3' group II intron fragment, a 5' group II intron fragment, and a protein-coding region or a non-coding region, and the nucleic acid molecule can be used to obtain a cyclic RNA having a biological activity. Further provided in the present disclosure are a vector containing the nucleic acid molecule, and the use thereof. A self-splicing system is constructed and optimized in the present invention, which determines that other group II intron self-splicing systems can be used in vivo and in vitro to generate a circular RNA and express an exogenous protein. The system demonstrates relatively high circularization efficiency and protein expression levels.

The application discloses a recombinant getah virus expressing E protein of Japanese encephalitis virus, the virus has the E protein gene of the Japanese encephalitis virus, and the E protein gene is located between Cap and E3 genes of the genome of the getah virus. A corresponding construction method is also established, the getah virus is used as a parent strain, an infectious clone pGETV-GV thereof is used as a carrier, a truncated E protein of the Japanese encephalitis virus is inserted between Cap and E3 by using a genetic engineering method, and the recombinant getah virus rGECJEV-E is obtained. Tests show that the recombinant getah virus rGECJEV-E has the characteristics of good performance, good stability and high expression amount of exogenous protein, the expressed protein has good immunogenicity, can provide immunoprotection for the attack of the Japanese encephalitis virus, and the safety of the recombinant strain is relatively high. Therefore, the recombinant virus has great potential in preparation of a vaccine or a medicine for preventing or treating the getah virus and / or the Japanese encephalitis virus, and is expected to develop a recombinant bivalent vaccine for the getah virus and the Japanese encephalitis.

The application relates to an anti-apoptosis baculovirus expression vector, which realizes a broad-spectrum anti-apoptosis effect by expressing siRNA targeting a Sf-caspase-1 and Tn-caspase-1 common sequence of Spodoptera frugiperda and Trichoplusia ni insect cells through the vector. The baculovirus vector contains a specific DNA sequence, the DNA sequence contains an siRNA sequence targeting the Sf-caspase-1 and Tn-caspase-1 common sequence transcribed by an RNA polymerase III promoter; the recombinant virus of the vector can express double-stranded small RNA in a host cell, silences the caspase-1 encoded by the host cell through an RNA interference pathway, thereby inhibiting the apoptosis of the host cell, and significantly improving the expression level of an exogenous protein. The application can be used for industrialized production of protein preparations and vaccines.