50 results about "Fusion Protein Expression" patented technology

Fusion proteins containing a targeting sequence, an exosporium protein, or an exosporium protein fragment that targets the fusion protein to the exosporium of a Bacillus cereus family member are provided. Recombinant Bacillus cereus family members expressing such fusion proteins are also provided. Genetically inactivated Bacillus cereus family members and recombinant Bacillus cereus family members that overexpress exosporium proteins are also provided. Seeds coated with the recombinant Bacillus cereus family members and methods for using the recombinant Bacillus cereus family members (e.g., for stimulating plant growth) are also provided. Various modifications of the recombinant Bacillus cereus family members that express the fusion proteins are further provided. Fusion proteins comprising a spore coat protein and a protein or peptide of interest, recombinant bacteria that express such fusion proteins, seeds coated with such recombinant bacteria, and methods for using such recombinant bacteria (e.g., for stimulating plant growth) are also provided.

Provided herein are fusion proteins in which IL-15 is tethered to receptor IL-2Rβ with an intervening linker, optionally a C-myc tag and / or IL-2Ra (CD25) extracellular domain linker, further optionally comprising an IL-2 signal peptide. Also provided are compositions comprising the fusion proteins, cells expressing the fusion proteins, and methods of treatment (e.g., of cancer) using the fusion proteins or cells.

The invention provides a plasmid for purine base mutation in blue-green algae and a preparation method and application thereof, and belongs to the field of bioengineering, the plasmid provided by the invention is named as pCyABE, the plasmid comprises a TadA-nCas9 fusion protein expression cassette and an sgRNA expression cassette, the nucleotide sequence of the TadA-nCas9 fusion protein expression cassette is shown as SEQ ID No.2, the nucleotide sequence of the sgRNA expression cassette is shown as SEQ ID No.2, and the nucleotide sequence of the sgRNA expression cassette is shown as SEQ ID No.2. The nucleotide sequence of the sgRNA expression cassette is as shown in SEQ ID No. 3. The plasmid provided by the invention is particularly suitable for mutation of adenine at any site on a blue-green algae genome into guanine, realizes fixed-point editing of the gene, has the advantages of high editing efficiency and low off-target rate, and has important significance on physiological property research and industrial application of blue-green algae.

Fusion proteins containing a targeting sequence, an exosporium protein, or an exosporium protein fragment that targets the fusion protein to the exosporium of a Bacillus cereus family member are provided. Recombinant Bacillus cereus family members expressing such fusion proteins are also provided. Genetically inactivated Bacillus cereus family members and recombinant Bacillus cereus family members that overexpress exosporium proteins are also provided. Seeds coated with the recombinant Bacillus cereus family members and methods for using the recombinant Bacillus cereus family members (e.g., for stimulating plant growth) are also provided. Various modifiations of the recombinant Bacillus cereus family members that express the fusion proteins are further provided. Fusion proteins comprising a spore coat protein and a protein or peptide of interest, recombinant bacteria that express such fusion proteins, seeds coated with such recombinant bacteria, and methods for using such recombinant bacteria (e.g., for stimulating plant growth) are also provided.

A recombinant humanized collagen type III alpha-1 (rhCol III a1), and an expression vector and use thereof are provided, relating to the technical field of bioengineering. A sequence on positions 154 to 1,221 of a human type III collagen α chain is ligated to an optimized vector pcDNA3.1, a resulting optimized expression vector is transfected into a human embryonic kidney cell Expi293F, and then rhCol III a1 is secreted to form a fusion protein with a soluble green fluorescent protein (GFP). The expressed rhCol III a1 that forms the fusion protein with the GFP shows an activity of promoting the migration of BALB / c 3T3 cells, thus exhibiting a great application potential in the field of skin care.

The invention relates to the technical field of biological medicines, and discloses a construction method and expression application of an Hcp gene deleted strain of a rahnella aqua KCL-5 strain for fish. The Hcp 0186 gene deletion strain is constructed by using a homologous recombination method, the comparison between the deletion strain and a wild strain is carried out, and the result shows that the deletion of the Hcp 0186 gene has an influence on the growth speed, the pathogenicity of the Hcp 0186 gene to the zebra fish is obviously reduced, the survival rate of the infected zebra fish is increased from 0% to 52%, and the Hcp 0186 gene deletion strain can be used as a fish attenuated live vaccine. A recombinant prokaryotic expression vector pET32a-Hcp and expression and purification of fusion protein of the recombinant prokaryotic expression vector pET32a-Hcp are further constructed, and Hcp 0186 rabbit polyclonal antibody serum is prepared as a primary antibody and can be applied to localization expression analysis of fish tissues infected by the bacterium and detection of WB and IHC. Results prove that the Hcp 0186 gene deletion of the strain provides an attenuated strain vaccine and clinical diagnosis and detection application of infected fish of the attenuated strain vaccine, and a scientific basis can be provided for revealing a molecular pathogenic mechanism of the strain and development and application of a mutant strain of the strain.

Provided are a fusion protein, a preparation method therefor and the use thereof. The fusion protein contains a His tag, a SUMO tag, and natural canine interferon-α, wherein the His tag has an amino acid sequence as represented by SEQ ID NO: 11, the SUMO tag has an amino acid sequence as represented by SEQ ID NO: 12, and the natural canine interferon α has an amino acid sequence as represented by SEQ ID NO: 7. The fusion protein exhibits an extremely significant increase in expression level and yield, has preparation stability and biological activity significantly greater than those of a native canine interferon-α protein, and has good safety.

The invention discloses a fusion protein expression system suitable for expressing long-chain helper protein in yeast, which is a gene expression cassette of fusion protein of helper protein and bovine lactoferrin, and comprises a double KREAEA structural domain + flexible Linker tandem helper protein gene and bovine lactoferrin gene, and the fusion protein assists in folding of bovine lactoferrin in cells, so that the expression of long-chain helper protein is realized. The fusion protein is excised in cells, and the secretion efficiency of correctly folded bovine lactoferrin is remarkably enhanced.

The invention belongs to the technical field of protein engineering, particularly relates to a protein tag with superoxide dismutase activity and application, and provides a protein tag which comprises polypeptide or protein with superoxide dismutase activity. On the second aspect, a fusion protein is provided and comprises the protein tag described in the first aspect and at least one target polypeptide or protein, the target polypeptide or protein is not superoxide dismutase, and the target polypeptide or protein is not superoxide dismutase only by using the protein tag disclosed by the invention. According to the present invention, the technical effects of target polypeptide or protein expression quantity detection (SOD enzyme activity detection), target polypeptide or protein protection from oxidative damage, target polypeptide or protein stability improvement and target polypeptide or protein solubility improvement can be simultaneously achieved without adding any additional step compared with the fusion protein expression operation in the prior art; the operation is convenient.

The invention discloses a swine acute diarrhea syndrome coronavirus subunit vaccine as well as a preparation method and application thereof. The method comprises the following steps: firstly, predicting the antigen epitope of SADS-CoV S1 protein through bioinformatics, synthesizing and screening the obtained antigen epitope peptide in vitro, and further screening the antigen epitope with high immunogenicity through a Dot-blot experiment. And then, constructing a recombinant fusion protein expression vector by using the SADS-CoV S1 gene and the screened antigen epitope peptide coding gene, and preparing the recombinant fusion protein after the recombinant plasmid is transformed into escherichia coli. A vaccine prepared by emulsifying the recombinant fusion protein by using an adjuvant is used for immunizing a mouse to evaluate the immunogenicity of the mouse, detection results of methods such as indirect ELISA and splenic lymphocyte proliferation experiments prove that the vaccine can effectively induce specific humoral immunity and cellular immunity effects, and the antigen epitope has the effect of enhancing the immunogenicity of the SADS-CoV S1 protein. The vaccine has a wide application prospect and can be used for preventing the acute diarrhea syndrome of pigs.

The present invention discloses a preparation method and application of a soluble B lymphocyte stimulator receptor 3 and adiponectin globular domain fusion protein. The present invention belongs to the field of biomedical technology. The preparation method includes the following steps: cloning the coding sequences of the extracellular region of BAFF-R3 and the adiponectin globular domain; connecting the extracellular region of BAFF-R3 and the adiponectin globular domain through a peptide chain, and inserting the expression sequence of the eBR3-gAD fusion protein into a plasmid using restriction enzymes and DNA ligase; transfecting the above plasmid into cells and screening out a cell line with stable expression; culturing the above cell line, collecting the supernatant, and purifying by gel filtration and chromatography to obtain the soluble B lymphocyte stimulator receptor 3 and adiponectin globular domain fusion protein. The preparation method of this novel fusion protein has the advantages of high specificity, high expression level, and simple purification.

The invention relates to the technical field of gene engineering, in particular to a fusion protein expression cassette for rice multi-gene editing and application of the fusion protein expression cassette. The expression of the fusion protein for multiple gene editing of rice provided by the invention comprises an endonuclease Csy4, a DNA (Deoxyribose Nucleic Acid) endonuclease nickase Cas9, an M-MLV reverse transcriptase and a Cas12a nuclease; in the guide editing fusion protein frame, the ribonuclease Csy4 is located at the N end of the guide editing fusion protein frame, the DNA incision enzyme nickase Cas9 is located between the ribonuclease Csy4 and the M-MLV reverse transcriptase, the M-MLV reverse transcriptase is located between the DNA incision enzyme nickase Cas9 and the Cas12a nuclease, and the Cas12a nuclease is located at the C end of the fusion protein frame. The fusion protein expression cassette not only can ensure the editing efficiency, but also can improve the editing accuracy.

This invention belongs to the field of plant gene editing technology, specifically relating to a bifunctional gene editing system and its application in single-parent maize. The invention provides a bifunctional gene editing system comprising: a fusion protein expression cassette and a guide RNA combined expression cassette; wherein the guide RNA combined expression cassette includes: pegRNA associated with the maize D9 gene; and sgRNA associated with the maize YUC4 and YUC2 genes. Transient expression validation using the bifunctional gene editing system in maize protoplasts revealed that the system achieves a precise editing efficiency of 3.5% at the G571V site of the D9 gene, a knockout efficiency of 40%-65% for the YUC4 gene, and a knockout efficiency of 37%-54% for the YUC2 gene. This demonstrates that the technical solution of this application can simultaneously and efficiently achieve precise point mutation and gene knockout within the same cell, thus enabling its application in the synergistic improvement of lodging resistance in maize.

A derivative of insulin aspart and a method for preparing the same, the derivative comprising a fusion protein of a green fluorescent protein folding unit and an insulin aspart precursor or an active fragment thereof. The fusion protein is expressed in a significantly increased amount, the insulin aspart precursor protein in the fusion protein is folded correctly and has biological activity. Furthermore, the green fluorescent protein folding unit in the fusion protein can be digested by a protease into small fragments, and has a large difference in molecular weight compared to the target protein, and is easy to separate. A method for preparing insulin aspart and an intermediate using the fusion protein is also provided.

The invention provides a plasmid tool for simulating Kras random evolution and application thereof, and the plasmid tool comprises at least two guide RNA expression cassettes driven by a first promoter, which respectively target a Pten gene and a Trp53 gene; the pilot editing guide RNA expression box is driven by a second promoter, targets the 12th codon of the Kras gene, and comprises a repair template sequence for editing the codon into G12D; the fusion protein expression cassette is driven by a third promoter and comprises SpCas9 protein and MMLV reverse transcriptase; and at least one eukaryotic selection marker gene expression cassette. The invention also discloses a method for constructing a simulated Kras random evolution model by adopting the plasmid tool, and application of the plasmid tool in screening targeted drugs aiming at KRAS mutation tumors. The plasmid tool provided by the invention can realize cell multi-gene locus editing, construct a Kras locus random evolution model and simulate a Kras random evolution process in ovarian cancer.

The invention provides a recombinant bovine lactoferrin peptide capable of reducing feedback inhibition on host bacteria and a construction method thereof in order to overcome the practical application problem of a genetic engineering method in the process of producing the bovine lactoferrin peptide at present. According to the invention, bovine lactoferrin peptide, calmodulin and enhanced green fluorescent protein are connected in series to form bovine lactoferrin peptide fusion protein, a pichia pastoris recombinant strain is constructed, and high-level expression of bovine lactoferrin peptide is realized. According to the method, positive charges of bovine lactoferrin are neutralized on the basis of calmodulin, toxicity of antibacterial peptide to a host is weakened, the tandem enhanced green fluorescent protein can emit green fluorescence under 488 nm exciting light, and real-time analysis of the expression level of bovine lactoferrin fusion protein is achieved. According to the method, by adopting a fusion expression technology, the toxicity to a host when the antibacterial peptide is produced by a genetic engineering method is overcome, so that the expression quantity is increased, and a foundation is laid for high-density fermentation production of the bovine lactoferrin peptide.

The invention provides a fusion protein of flounder netshell virus nucleocapsid protein, bacillus subtilis for expressing the fusion protein and an oral vaccine, and relates to the technical field of biology. According to the present invention, the sequence of the bacillus subtilis anchoring protein CotC is obtained through sequencing, the bacillus subtilis anchoring protein CotC and the flounder nested virus are fused, and after the fusion protein is introduced into the bacillus subtilis, the fusion protein can be anchored on the surface of the bacterial spore, and can be used for preparing the recombinant oral vaccine capable of being stably stored and directly fed, and a new method is developed for preventing the flounder netlike virus CSBV.

The invention discloses methods for precision editing of a highly homologous region. The method includes identification of a region of interest for precision editing of the homologous regions. A vector construct for expression of a fusion protein specific to the region of interest is obtained. The fusion protein is a gene construct having a sequence for expressing deadCas9 nuclease and a ABE8e base editor for editing at the identified region of interest. One or more vector constructs are created for expression of guideRNA complementary to the region of interest. The gRNA vector construct along with the fusion protein expression vector are transformed into a suitable cell line for precision editing of the highly homologous regions. The invention further discloses another method for precision editing of a highly homologous region by using truncated guideRNAs having a base pair length of 10 -15 nucleotides for precision editing in highly homologous regions.

The present invention provides a nanobody-assisted protein or polypeptide expression system, and preparation and use, and specifically relates to a nanobody fusion protein, an expression vector, a host cell and a nanobody-assisted protein expression system for promoting protein or polypeptide expression and secretion, and preparation and use thereof.

The invention relates to the technical field of synthetic biology, and particularly discloses an artificially designed chlamydomonas reinhardtii chloroplast genome as well as an assembly method and application thereof. Wherein the artificially designed chlamydomonas reinhardtii chloroplast genome is obtained by modifying a simplified chlamydomonas reinhardtii chloroplast genome, and the method comprises the following steps: deleting repetitive sequences located at the 27767-44496 sites of the simplified chlamydomonas reinhardtii chloroplast genome, and the simplified chlamydomonas reinhardtii chloroplast genome is derived from chlamydomonas reinhardtii with the preservation number of CCTCC (China Center For Type Culture Collection) NO.M20221032. According to the application, a segment of long repetitive sequence as long as 17031 bp in a simplified Chlamydomonas reinhardtii chloroplast genome is deleted, the genome structure is simplified, and meanwhile, a LacO sequence and a LacI-mVenus fusion protein expression cassette are added, so that the chloroplast genome has a visualization function, and the artificially designed Chlamydomonas reinhardtii chloroplast genome has better stability and functional performance.

The invention discloses an expression method of a non-proteolytic thrombin peptide TP508 in bombyx mori silk gland and a product and application thereof, a fusion expression vector of the non-proteolytic thrombin peptide TP508 and a red fluorescent protein DsRED is constructed, the fusion protein is expressed in silkworm cocoon silk, the bred transgenic strain silk successfully expresses the fusion protein, and the non-proteolytic thrombin peptide TP508 is obtained. The thrombin peptide TP508 fusion protein expressed by the transgenic silk not only can remarkably promote cell proliferation, cell migration and vascularization, but also keeps the characteristic of red fluorescence, and is expected to be applied to the fields of biomedicine and tissue cell engineering.

The invention provides a nano-antibody-assisted protein or polypeptide expression system as well as preparation and application thereof, and particularly relates to a nano-antibody fusion protein for promoting protein or polypeptide expression and secretion, an expression vector, a host cell and a nano-antibody-assisted protein expression system as well as preparation and application thereof.

The invention discloses an antigen-loaded PHA (polyhydroxyalkanoate) nano-particle expression plasmid system as well as preparation and application of the antigen-loaded PHA nano-particle expression plasmid system. Comprising a pACYC-PhaA-PhaB plasmid and a pET28a-PhaC-Target plasmid, and the nucleotide sequences of the pACYC-PhaA-PhaB plasmid and the pET28a-PhaC-Target plasmid are respectively shown as SEQ ID NO: 4 and SEQ ID NO: 5; the pACYC-PhaA-PhaB plasmid is characterized in that a PhaA protein expression cassette and a PhaB protein expression cassette, which are respectively started by two T7 promoters, are inserted into multiple cloning sites of a vector pACYCDuet-1; the pET28a-Phac-Target plasmid is characterized in that a PhaC-vaccine antigen fusion protein expression cassette is inserted into a plurality of cloning sites of a vector pET-28a. The invention lays a good foundation for preparation of PHA nanoparticle vaccines and related research.

The invention belongs to the technical field of environmental detection and biology, and particularly discloses a cascade amplification gene expression regulation device or system based on a GAL4 / UAS system and application of the cascade amplification gene expression regulation device or system based on the GAL4 / UAS system. An estrogen receptor reaction element ERE responds to an estrogen signal, GAL4 fusion protein expression is driven, an upstream activation sequence UAS is combined to form a transcription complex, and the GAL4 / UAS cascade amplification gene expression regulation device or system based on the GAL4 / UAS system is obtained. And the expression of a downstream reporter gene (such as mScarlet3) or a target gene is amplified in a cascade manner. The system realizes high-sensitivity estrogen detection (the detection limit reaches 10 <-1 > pM) in mammalian cells, and an environmental estrogen biosensor can be constructed. The kit comprises a regulation and control system and a transfection reagent, is suitable for quantitatively monitoring fluorescence intensity change so as to evaluate estrogen activity, and has the technical advantages of high efficiency, sensitivity and modularization.

The present invention relates to the fields of cancer and immunotherapy, and specifically to CD80 IgV variant polypeptides, CD80 ECD variant polypeptides, CD80 variant fusion polypeptide complexes comprising these variant polypeptides, and nucleic acid molecules that express these variant fusion polypeptide complexes as fusion proteins. The present invention also relates to methods of inducing or enhancing immunity and methods of treating diseases such as infectious diseases and cancer using these variant polypeptides, fusion polypeptide complexes, or nucleic acid molecules.

Disclosed herein are methods for precise editing of highly homologous regions. The method includes identifying a target region for precise editing of a homologous region. A vector construct for expressing a fusion protein specific for the target region is obtained. The fusion protein is a genetic construct having sequences for expressing an inactivated Cas9 nuclease and an ABE8e base editor for editing at the identified target region. One or more vector constructs for expressing a guide RNA complementary to the target region are created. The gRNA vector construct(s) are transformed into a suitable cell line along with the fusion protein expression vector for precise editing of the highly homologous region. Also disclosed herein is another method for precise editing of highly homologous regions using a truncated guide RNA of 10-15 nucleotides in length for precise editing in highly homologous regions.

Adeno-associated virus (AAV) gene therapy vectors express a therapeutic protein or RNA that treats a genetic defect in a cell. The present disclosure provides AAV gene therapy vectors that additionally express an anti-inflammatory protein or peptide. When the provided AAV gene therapy vectors are used in methods of treatment of. for example. neurodegenerative diseases. the therapeutic protein / RNA treats the genetic defect within the cells directly transduced by the AAV vectors while the anti-inflammatory protein / peptide is secreted by the transduced cells into the intercellular milieu and treats microglial activation associated with the neuroinflammatory component of the neurodegenerative diseases. The therapeutic protein and anti-inflammatory protein / peptide can be expressed as fusion protein in which the two are separated in the fusion protein by a self-cleaving peptide. The provided AAV gene therapy vectors and methods are thus useful in treating neurological and neurodegenerative disorders.

The invention relates to the field of plant genetic engineering, and particularly discloses a recombinant vector construction system for tobacco eIf4e gene editing. The system comprises a recombinant expression vector with pCAMBIA1300 as a basic skeleton, an nCas9-RT fusion protein expression unit driven by a Stubi promoter and a pegRNA expression unit driven by an NtU6-28 promoter are integrated in the recombinant expression vector, and the system further comprises a hygromycin resistance selection marker driven by a 35S promoter; the preparation method comprises the following steps: firstly, synthesizing a specific nucleotide sequence of each element, carrying out directional cloning through BstBI and BlpI restriction endonuclease sites, and sequentially connecting the two core expression units in series according to the direction from 5'to 3 'and assembling the two core expression units into a vector skeleton. The recombinant vector system can be used for specific multi-site precise editing of a VGAD region of the tobacco eIf4e gene, and has the advantages of dual-promoter coordinated regulation and editing element expression coordination.

The application discloses an expression plasmid system of antigen-loaded PHA nanoparticles and preparation and application thereof. The expression plasmid system comprises pACYC-PhaA-PhaB plasmid and pET28a-PhaC-Target plasmid, nucleotide sequences of the plasmid and the pET28a-PhaC-Target plasmid are respectively shown as SEQ ID NO: 4 and SEQ ID NO: 5; the pACYC-PhaA-PhaB plasmid is a PhaA and PhaB protein expression frame inserted into a multiple cloning site of a vector pACYCDuet-1 and driven by two T7 promoters respectively; the pET28a-Phac-Target plasmid is a PhaC-vaccine antigen fusion protein expression frame inserted into a multiple cloning site of a vector pET-28a. The application lays a good foundation for preparation of PHA nanoparticle vaccines and related research.