80 results about "Gene coexpression" patented technology

The invention provides polynucleotides and polypeptides that are co-expressed with genes known to be involved in bone remodeling and osteoporosis. The invention also provides expression vectors, host cells, and methods for making a polypeptide, for screening or purifying ligands, and for diagnosing disorders associated with bone remodeling or osteoporosis.

The invention belongs to the field of data excavation and discloses a protein functional module excavating method for multi-view data fusion. The method comprises the following steps: firstly performing quantifying description on strong and weak interaction of multiple data sources on protein and forming multi-view data; further performing uniform matrix decomposition on the multi-view data by utilizing a polymerization nonnegative matrix algorithm provided by the invention; determining the functional module of the protein by virtue of obtaining the optimal approximation of the multi-view information. The protein functional module excavating method for multi-view data fusion, provided by the invention, aims at simultaneously analyzing multiple biodata and comprises gene coexpression, GO annotation and PPIN and can be used for extracting the protein functional module with the most consistent polymerization characteristic from the multi-view. The method disclosed by the invention is especially suitable for interaction networks and biodata of the protein and meanwhile can be applied to community excavation problems of social complex networks and communication networks.

The invention provides construction of an EP153R and EP402R gene co-expression recombinant adenovirus vector and a adenovirus packaging method, and belongs to the technical field of genetic engineering. Based on a pShuttle-CMV eukaryotic expression vector, an EP153R gene and EP402R gene overexpression adenovirus vector pAD-Shuttle-CMV-CTLA4-EP153R-EP402R is introduced with CTLA4, EP153R and EP402Rgenes at multiple cloning sites. The invention also provides recombinant adenovirus for co-expressing EP153R and EP402R genes, and the adenovirus packaging process is achieved by utilizing the constructed pAD-Shuttle-CMV-CTLA4-EP153R-EP402R vector to obtain the adenovirus capable of directly infecting animal or eukaryotic cells, thereby achieving the purpose of co-expressing EP153R and EP402R genes in eukaryotic cells, and laying a foundation for researching the adenovirus vaccine for achieving the co-expression of EP153R and EP402R genes.

The invention relates to the technical field of bioinformation, and discloses a marking method and system for protein functions. Therefore, protein marking performance is improved, expensive cost of a bioexperiment method and poor efficiency are solved. The method comprises following steps: estimating the first possibility of a certain function in a to-be-inquired protein according to a first-stage structure neighborhood and a second-stage structure neighborhood; estimating the second possibility of the certain function in the to-be-inquired protein according to all homologous sequences; inputting a PSSM matrix of the to-be-inquired protein into an SVM prediction model to obtain the third possibility of the certain function in the to-be-inquired protein; converting the distribution of the function corresponding to other species according to the gene co-expression fraction into the fourth possibility of the function occurring in a target species in the to-be-inquired protein; and mixing the first possibility, the second possibility, the third possibility and the fourth possibility to estimate the comprehensive possibility of the function in the to-be-inquired protein.

The invention provides a method for establishing a recombinant adenovirus vector with Africa swine fever EP153R and P54 gene coexpression and packaging adenovirus and belongs to the technical field ofgene engineering. According to the adenovirus vector of coexpression of the EP153R gene and the P54 gene, with a pShuttle-CMV eukaryotic expression vector as a basis, the CTLA4, EP153R and P54 genesare introduced at multiple cloning sites. The invention further provides recombinant adenovirus with coexpression of the EP153R and P54 genes. The established pAD-Shuttle-CMV-CTLA4-EP153R-P54 vector is used for achieving the adenovirus packaging process, the adenovirus which can directly infect animals or eukaryocyte is obtained, the aim of coexpression of the EP153R and P54 genes in eukaryocyte is achieved, and the basis is laid for research on adenovirus vaccines of coexpression of the EP153R and P54 genes.

The invention relates to the technical field of gene engineering, in particular to an immune enhanced gene vaccine of a hepatitis B virus core antigen (HBcAg). The vaccine is an immune stimulating complex (ISCOM) vaccine consisting of HBcAg and OX40 ligand (OX40L) dual-gene co-expression recombinant eukaryotic vector, saponin Quil A, cholesterol and lecithin. The vaccine can promote immune response of specific T cells of the HBcAg and efficiently induce the anti-virus replication capacity of immune cells, and has good application prospect in the field of hepatitis B virus infection resistance. The invention also relates to a preparation method for the vaccine with simple and convenient operation and low cost.

The invention relates to a bicistronic mRNA coexpression gene transporter and a preparation method thereof. The gene order of the gene transporter is SEQ ID No. 15; the gene transporter from 5' end to 3' end comprises bovine Nuclear matrix binding region MAR, artificially constructed combined promoter CAG, Profilin gene FSTN, internal ribosome entry site (IRES), green fluorescent protein AcGFP gene and Rabbit globin poly A signal region; the preparation method comprises the following steps: constructing a carrier pCAG-IRES2-AcGFP1; obtaining FSTN gene and inserting the pCAG-IRES2-AcGFP1 carrier; obtaining the sequence of MAR and inserting the pCAG-IRES2-AcGFP1 carrier; constructing a plasmid carriers so as to obtain the bicistronic mRNA coexpression gene transporter. The gene transporter is not only pure and safe gene transporter but also realizes dual-gene coexpression in any combination, achieves the purpose of multi-gene coexpression through repeated utilization of IRES, and provides new thinking and route for improving the shape of dual or multiple gene control such as economic character.

The invention relates to a human Bcl-2 and human VEGF165 double-gene co-expression recombinant vector and a building method thereof, and is characterized in that IRES segment is utilized to connect an hBc1-2 gene and an hVEGF165 gene, and homologous recombination mechanisms in bacterium are utilized to recombine and build the human Bcl-2 and human VEGF165 double-gene co-expression recombinant vector according to an AdEasy system. The invention leads the Bcl-2 gene with anti-apoptosis capacity and the VEGF165 gene which currently promotes angiogenesis cytokine with strongest function to recombine on a same vector to be co-expressed, and solves the problem of low survival rate of transplanted cells under the condition of hypoxia and inflammation, simultaneously plays the function of promoting angiogenesis of VEGF165, plays an important part in the gene therapy of myocardial infarction and the cell transplantation research field, and has wide application prospect.

The invention discloses a production method of recombinant NotI restriction endonuclease in escherichia coli. The production method comprises the following steps of a) construction of a pCreat Duet-NotI-methylase expression plasmid, b) induction expression and identification of NotI restriction endonuclease and methylase protein, and c) purification and identification of the NotI restriction endonuclease and the methylase protein. According to the production method, an escherichia coli expression system is applied, a coexpression vector is designed for coexpression with a methylase gene, and methylase can protect an escherichia coli genome against an influence of NotI, so that expression of the recombinant NotI in the escherichia coli is realized; purification is performed on NotI protein by affinity chromatography and Superdex 200 gel filtration chromatography and a whole purification process needs 5-6h, so that a simple two-step purification manner greatly shortens a purification procedure and the purification time of the NotI protein, and the purity, a yield and the enzyme activity of the NotI protein are improved and increased; and a novel method of a purification technology of the NotI protein lays a foundation for researching, developing and producing other II-type restriction endonuclease.

The invention discloses an AFP and GM-CSF dual-gene co-expression recombinant vector, which is sequentially linked with an AFP gene, an IRES sequence and a GM-CSF gene in a vector transcription direction, or is sequentially linked with a GM-CSF gene, an IRES sequence and an AFP gene in a vector transcription direction, wherein nucleotide sequence of the AFP gene is represented in SEQ ID NO: 1 in a sequence list, nucleotide sequence of the GM-CSF gene is represented in SEQ ID NO: 2 in the sequence list, and nucleotide sequence of the IRES sequence is represented in SEQ ID NO: 3 in the sequence list. The dual-gene co-expression recombinant vector, which links the AFP gene and the GM-CSF gene through the IRES sequence, can simultaneously express alpha fetal protein and a granulocyte-macrophage colony stimulating factor in a same vector; and the recombinant vector, in immunogene therapy of liver cancer, not only can develop an immune regulating function of a cell factor but also can generate a specific anti-tumor effect targeted for the liver cancer.

The invention discloses construction of a three-gene co-expression vector for efficiently synthesizing tetrahydropyrimidine and application of the co-expression vector. The construction method comprises the following steps: sequentially inserting coded L-diaminobutyrate aminotransferase (ectA) gene,coded L-diaminobutyric acid acetyltransferase (ectB) gene and coded tetrahydropyrimidine synthase (ectC) gene in pseudo-bacillus firmus into a transformed plasmid pET-22bNS according to an isocaudarner principle, and constructing the three-gene tandem co-expression vector pET-22bNS-EctA/B/C. When the constructed three-gene co-expression vector is transferred into Escherichia coli BL21 (DE3) and is subjected to IPTG induction, after a recombinant strain whole-cell catalysis reaction for 3 hours, the highest yield of tetrahydropyrimidine is 9.8mg/mL, and the synthetic efficiency is 78.4mg/mL/d and is higher than a synthesizing level in similar researches. The three-gene co-expression vector constructed by the invention has the ability of efficiently synthesizing the tetrahydropyrimidine and has an excellent application value.

The invention relates to genomics, molecular biology, cell biology, gene engineering and clinical medicine, in particular to application of a human ING4 and IL-24 double-gene coexpression vector as a radiotherapy sensitizing agent. Before the radiotherapy, the human ING4 and IL-24 double-gene coexpression vector is led in tumor cells so that the human ING4 and IL-24 genes are expressed in the tumor cells, and then the radiotherapy is carried out. The Ad-ING4-polyA<delta 296-298>+CMV-IL-24 can retard lots of tumor cells in the proliferative stage in the G2/M stage and is beneficial to enhancing the irradiation sensitivity. The MTT and the FCM detection results further show that the Ad-ING4-IL-24 combined radiotherapy has the effect of obviously inhibiting the growth of the SPC-A1 lung adenocarcinoma cells, MDA-MB-231 breast carcinoma cells and the transplanted tumor thereof and inducing the cell apoptosis, exceeds the Ad-ING4-IL-24 single genome and the radiotherapy single genome, and presents obvious radiotherapy sensitization synergistic effect. Therefore, the human ING4 and IL-24 double-gene coexpression vector can be used for enhancing the sensitivity of the tumor cells to the radiotherapy.

The invention discloses an AFP (Alpha Fetoprotein) and IL-2 (Interleukin-2) double-gene co-expressing recombinant vector, wherein an AFP gene, an IRES (Internal Ribosome Entry Site) sequence and an IL-2 gene are sequentially connected along a transcription direction of the vector, or the IL-2 gene, the IRES sequence and the AFP gene are sequentially connected along the transcription direction of the vector; the nucleotide sequence of the AFP gene is shown as SEQ ID NO:1 in a sequence table, the nucleotide sequence of the IL-2 gene is shown as SEQ ID NO:2 in the sequence table, and the nucleotide sequence of the IRES sequence is shown as SEQ ID NO:3 in the sequence table. According to the double-gene co-expressing recombinant vector disclosed by the invention, the AFP gene and the IL-2 gene are connected via the IRES sequence, so that the AFP gene and the IL-2 gene can be simultaneously expressed in the same vector; the recombinant vector can be applied to the genetic immunization treatment of liver cancer, thereby achieving an immunomodulatory effect of cell factors and producing a specific anti-tumor effect to liver cancer in a targeted manner.

The invention discloses a lung cancer prognosis comprehensive prediction model and a construction method and device. The method comprises the following steps: collecting original gene expression data and corresponding clinical survival data of a lung cancer sample and conducting data preprocessing and standardization, thereby acquiring a gene expression matrix; obtaining the types of immune cells in the tumor and calculating the relative ratio of the various types of immune cells; screening out parameters for constructing a prognosis prediction model from the obtained immune cell types and obtaining corresponding regression coefficients, wherein the parameters are multiple immune cell types; based on the screened parameters, calculating an immune score according to the relative ratio and the corresponding regression coefficient; identifying a gene co-expression module, searching for a gene module cooperatively expressed in the lung cancer sample, and determining a gene module related to prognosis; and constructing a prognosis comprehensive prediction model. According to the invention, immune scores, clinical information and gene co-expression module characteristics are integrated to construct a comprehensive prediction model to predict prognosis of lung cancer patients.

The invention discloses a hepatitis B virus dendritic cell therapeutic vaccine and a preparation method thereof. The preparation method thereof includes the steps that: HBcAg-Fc digenic co-expression recombinant eukaryotic vector is constructed; the obtained digenic co-expression recombinant eukaryotic vector is added with saponin Quil A and then is added with cholesterol and lecithin, ice-bath ultrasonic processing is carried out, so that solution is fully mixed and emulsified, PBS dialysis is carried out, and the obtained micro floccus is immunostimulating complex; and the obtained immunostimulating complex activates and simulates dendritic cell, thus obtaining the cell therapeutic vaccine. The vaccine carries out high efficiency transfection on dendritic cell, the generated fusion protein targets the dendritic cell in high efficiency by virtue of Fc segment, thus activating the dendritic cell, promoting immune response of hepatitis B virus core antigen specificity T cell and inducing the antivirus replication capability of immune cell in high efficiency, so that the vaccine can be taken as hepatitis B virus therapeutic vaccine.

The invention relates to an antiapoptosis high expression hVEGF165 (human Vascular Endothelial Growth Factor 165) cell model and a building method thereof. The cell model takes BMSCs (Bone Mesenchymal Stem Cells) as transfection cells and coexpresses a recombinant adenovirus with double genes hBCL-2 and hVEGF165. The building method of the cell model comprises the following steps of: 1) separating and purifying rat BMSCs to obtain massive purified BMSCs with good vitality; 2) transfecting the BMSCs by the recombinant adenovirus, building the cell model built by the BMSCs transfected for 72 hours by adopting a multiplicity of infection (MOI) is equal to 400. The application characteristics are as follows: 1) the cell model is used for treating diseases such as myocardial infarction and the like; and 2) the cell model is used for being transplanted into a receptor organ. Compared with common BMSCs, the transfection recombinant adenovirus BMSCs provided by the invention has the advantages that cell apoptosis ratio is obviously reduced, service life is prolonged, the hVEGF165 can be efficiently expressed, the cell model can be conveniently transplanted to the receptor organ, and effect for treating diseases can be improved, thus the cell model and the building method thereof which are provided by the invention have wide application prospect.

The invention discloses extracellular expression and application of tyrosine phenol lyase and belongs to the technical field of biology. A method for extracellular expression includes: cloning high-activity tyrosine phenol lyase gene, co-expressing with signal peptide signal, converting Escherichia coli competent cells, and building a tyrosine phenol lyase extracellular secretion engineering strain mediated by signal peptide to realize extracellular expression, wherein obtained tyrosine phenol lyase fermentation liquid is used for converting and synthesizing tyrosine and derivatives. An extracellular expression mode is adopted to build the tyrosine phenol lyase engineering strain, and enzyme release quantity reaches higher than 50%, so that toxicity caused to cells due to excessive accumulation of protein in the cells is reduced, and total enzyme activity is improved by 36% when compared with common expression. Part of enzyme generated by fermentation is released into the fermentationliquid, thallus cells are high in permeability, contact of a substrate with enzyme is facilitated, and catalytic efficiency of the enzyme is improved substantially. The fermentation liquid can be directly adopted to catalyze the substrate to product L-tyrosine and the derivatives, and the cells do not need to be broken to extract pure enzyme for reaction, so that using cost of the enzyme is saved.

The invention discloses a building method of a tsh and iss double-gene coexpression strain. The method sequentially comprises the steps of double-gene clone, connection of tsh gene and iss gene with pETDuet-1 plasmids, expression strain conversion, target gene coexpression and the like. A microbial cell model based on tsh and iss is built; the biotransformation is performed; the method is simple;the process can be easily controlled; the double genes of tsh and iss are contained in the coexpression strain; the success ratio of the double-gene expression is high. The invention also provides application of the strain to prevention and treatment of avian colibacillosis. The method provided by the invention is applicable to the building of the tsh and iss double-gene coexpression strain; a gene engineering bacterium vaccine prepared from the strain is applied to the prevention and treatment of avian colibacillosis.

The invention relates to a construction method of a plant dual-gene co-expression vector. The construction method is characterized by comprising the following steps of: adding restriction enzyme cutting sites of Nco1 and EcoR I incision enzymes at two ends of a spacer sequence in a plasmid pGWB 605 (the registration number of which is AB543114 in GenBank) through PCR (Polymerase Chain Reaction) amplification, and then cloning the spacer sequence into a pXCS-HAStrep plasmid to construct the plant dual-gene co-expression vector pXCS-Dgene-HAStrep. The co-expression vector only has 5682Kb and can be loaded with two larger foreign genes which are independently expressed in an eukaryotic expression system. A bar gene is used as a screening marker gene in the co-expression vector. According to the construction method, the screening operation of a transgenic plant is simple, and the efficiency is high.

The invention provides an expression method and application of an ALB fusion protein, and the expression method comprises the following steps: cutting a genome by using nuclease, and simultaneously adding two fusion protein genes to an ALB gene through a gene co-expression connection element before a termination codon of the ALB gene of a human pluripotent stem cell or liver cell; the reporter gene is a fluorescent protein gene, a cell surface antigen, a luciferase gene, a chloramphenicol acetyltransferase gene, a beta-galactosidase gene, a secretory human placenta alkaline phosphatase gene or a PET reporter gene; human pluripotent stem cells are differentiated into liver cells through a directional differentiation technology. By adopting the technical scheme provided by the invention, the reporter gene or other functional genes can be efficiently connected with the ALB target gene, and the reporter gene or functional genes can be co-expressed with the target gene, so that specific reporter genes or functional proteins can be expressed in hepatocytes or hepatocyte-like cells.