112 results about "Foreign 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 biological medicine, and discloses lactococcus lactis HX21473 and application thereof, the lactococcus lactis HX21473 is preserved in China General Microbiological Culture Collection Center (CGMCC) on January 20, 2025, and the preservation number is CGMCC NO.33476. The invention further discloses a preparation method of the lactococcus lactis HX21473. The invention provides a strain of lactococcus lactis HX21473 capable of efficiently expressing a foreign protein. The lactococcus lactis HX21473 has strong acid resistance, can be used as an oral vaccine live carrier, and promotes research, development and application of oral vaccines.

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 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 belongs to the technical field of genes, and discloses a method for efficiently expressing foreign protein based on an NC048599.1 site in a CHO cell genome. A site for stably expressing a protein in a CHO cell genome is located in a range of 69195000 to 69199000 of a CHO cell gene NC048599.1 with a sequence as shown in SEQ ID NO: 1 (Sequence Identifier Number 1), and the sequence of the CHO cell gene NC048599.1 is shown in SEQ ID NO: 1. Different protein genes are introduced at fixed positions in a CHO cell genome, and stable expression is carried out.

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 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 relates to a Sebastes schlegeli endogenous retrovirus envelope protein Penv, a lentiviral vector and application thereof, and belongs to the field of genetic breeding of molecular biology, and the amino acid sequence of the envelope protein Penv is as shown in SEQ ID NO.1. The invention further provides an in-vitro assembly system, a transformant and a kit containing membrane fusion protein particles and lentiviral particles of the envelope protein Penv and application of the envelope protein Penv. VSVG protein is replaced with Penv protein from sclerobone fish, so that the transduction efficiency of the lentiviral particles to sclerobone fish cells is effectively improved, and the transduction efficiency of the lentiviral particles to the sclerobone fish cells is improved. And efficient gene delivery is realized. Meanwhile, as the endogenous Env protein, the Env protein overcomes the immunological rejection of a host to the exogenous Env protein, and also has the potential of in-vivo application.

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 invention belongs to the technical field of biological genetic transformation, and relates to a method for preparing recombinant foreign protein based on transient expression of fruits of cucurbitaceae crops. The efficient expression of the foreign protein is realized through the processes of constructing an instantaneous transformation vector, infecting fruits with agrobacterium and purifying the protein. The method overcomes the defects of high cost and low purity of a traditional tobacco transient expression system, has the advantages of easily available raw materials, large biomass, edibility, high safety and the like, and provides a new way for low-cost large-scale production of recombinant proteins.

The invention belongs to the technical field of biological medicine, and discloses lactococcus lactis HX9302 for expressing foreign protein and application, the lactococcus lactis HX9302 is preserved in China General Microbiological Culture Collection Center on January 20, 2025, and the preservation number is CGMCC NO.33477. The lactococcus lactis HX9302 is preserved in China General Microbiological Culture Collection Center on January 20, 2025; the lactococcus lactis has good acid resistance and digestive enzyme tolerance, can be used as an oral delivery live carrier of helicobacter pylori vaccines, and promotes research, development and application of oral vaccines.

The invention particularly relates to a sebastes schlegeli endogenous retrovirus envelope protein percomORF, a lentiviral vector and application of the sebastes schlegeli endogenous retrovirus envelope protein percomORF and the lentiviral vector, and belongs to the field of genetic breeding of molecular biology, and the envelope protein percomORF has an amino acid sequence as shown in SEQ ID NO.1. The invention also covers a membrane fusion protein particle constructed based on the protein, a system for in-vitro assembly of lentivirus particles, a corresponding transformant and a matched kit. By replacing conventional VSVG protein with percomORF protein from sclerobone fish, the infection and transduction efficiency of lentivirus on sclerobone fish cells can be effectively enhanced, and more efficient gene delivery is realized. Besides, the protein belongs to host endogenous Env protein and is beneficial to avoiding immunological rejection caused by introduction of foreign protein, so that the protein has good potential in in-vivo gene delivery application.

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 invention relates to the technical field of saccharomyces cerevisiae expression foreign protein, and discloses a saccharomyces cerevisiae strain with ASFV p14.5 displayed on the surface and a construction method and application thereof, and the method comprises the following steps: (1) synthesizing a delta site homologous arm sequence; (2) seamlessly connecting the homologous arm sequence of the delta site to a carrier pET23a, connecting a selective marker tryptophan biosynthetic gene TRP1 to the 5'end of the upstream homologous arm sequence of the delta site, and sequentially connecting an Aga2 gene, an E120R gene, a T2A peptide gene and an mCherry gene to the 5 'end; (3) carrying out enzyme digestion on two sides of the delta site through restriction endonuclease salI, and recovering a linearized fragment through agarose gel; and (4) integrating the recovered fragments into a host saccharomyces cerevisiae genome through a chemical conversion method, and culturing until bacterial colonies grow. The surface display type saccharomyces cerevisiae strain obtained by the invention not only has the stability of integrative saccharomyces cerevisiae for expressing foreign protein, but also can be used for more easily and more intuitively screening a high-expression quantity saccharomyces cerevisiae strain by combining the T2A peptide with mCherry.

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 relates to the technical field of medicine and biology, and particularly discloses a stress-inducible virus and application thereof.The stress-inducible virus comprises a stress-inducible adenovirus or a stress-inducible lentivirus, the nucleotide sequence of the stress-inducible adenovirus or the stress-inducible lentivirus is shown as SEQ ID NO: 1, and the nucleotide sequence of the stress-inducible adenovirus or the stress-inducible lentivirus is shown as SEQ ID NO: 2. The preparation method comprises the following steps: constructing a virus vector, wherein the virus vector has a nucleotide sequence as shown in SEQ ID NO: 1; and virus packaging: transfecting HEK293T cells by adopting shuttle plasmids containing SEQ ID NO: 1 and skeleton plasmids containing viral genome sequences, and collecting a virus stock solution to obtain the stress induced virus. The invention also discloses application of the stress-induced virus in preparation of medicines for preventing and treating oral tumors. Controllable release of the foreign protein under a specific signal is realized. The therapeutic protein produced by the invention can maximize the curative effect of drugs such as CD47 targeted protein and the like, and meanwhile, the toxic and side effects of the drugs on other tissues of the whole body are reduced.

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 present invention makes it possible to efficiently extract and purify a target foreign protein expressed using Escherichia coli from the periplasm.　A protein production method that includes steps (a)–(c).　(a) A step for introducing a gene that codes for a target foreign protein into Escherichia coli in which a gene associated with maintenance of outer membrane structure has been modified. (b) A step for culturing the Escherichia coli from (a). (c) A step for collecting the target foreign protein from the periplasmic fraction of the cultured bacterial cells.

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.

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.

The invention belongs to the technical field of biosynthesis, and particularly relates to an engineered microorganism, a construction method of the engineered microorganism, a recombinant vector and a production method of N-lactylphenylalanine. An exogenous YbdK gene and / or a coding gene for coding an exogenous YbdK protein are / is inserted into the engineered microorganism; the YbdK gene expresses the exogenous YbdK protein in the engineered microorganism, and / or the engineered microorganism expresses the exogenous YbdK protein, the YbdK gene has a nucleotide sequence as shown in SEQ ID NO: 1, the YbdK protein has an amino acid sequence as shown in SEQ ID NO: 2, and the engineered microorganism has increased yield of N-lactoyl phenylalanine. The engineered microorganism provided by the embodiment of the invention can provide the yield of N-lactoyl phenylalanine which is increased by about 15 times compared with a no-load control.