1427 results about "Fusion gene" patented technology

The invention relates to a preparation method for a vaccine capable of treating and/ or preventing SARS-CoV-2 infection or COVID-19 diseases. The core antigen of the vaccine comprises the RBD (receptor binding zone) fusion protein of the SARS-CoV-2, and a vaccine form comprises an RBD fusion protein subunit vaccine, an RBD fusion protein mRNA vaccine or an RBD fusion protein adenovirus vector vaccine. The above vaccine immunizes an organism, and immune reaction for treating and/ or preventing the SARS-CoV-2 infection can be generated so as to be used for treating and/ or preventing COVID-19. The invention also relates to an RBD fusion gene, the RBD fusion protein, a carrier, a cell, a preparation method, a treatment method or a pharmacy purpose of the SARS-CoV-2.

We have made retrovirus particles displaying a functional antibody fragment. We fused the gene encoding an antibody fragment directed against a hapten with that encoding the viral envelope protein (Pr80env) of the ecotropic Moloney murine leukemia virus. The fusion gene was co-expressed in ecotropic retroviral packaging cells with a retroviral plasmid carrying the neomycin phosphotransferase gene (neo), and retroviral particles with specific hapten biding activities were recovered. Furthermore the hapten-binding particles were able to transfer the neo gene and the antibody-envelope fusion gene to mouse fibroblasts. In principle, the display of antibody fragments on the surface of recombinant retroviral particles could be used to target virus to cells for gene delivery, or to retain the virus in target tissues, or for the construction of libraries of viral display packages.

The invention provides a genome editing method based on a CRISPR (Clustered Regularly Interspaced Short Palindromic Repeat) system and belongs to the technical field of gene editing. The genome editing method provided by the invention utilizes introns to express guide RNA molecules in the CRISPR genome editing system and adopts a principle that one or more than one tRNA-gRNA or crRNA series unitsare arranged in the introns of an encoded gene, and after the introns form a fusion gene with Cas9 or Cpf1, a promoter can be used for driving. The genome editing method provided by the invention hasthe benefits that a cellular endogenous mRNA splicing system and a cellular endogenous tRNA processing system are skillfully utilized, and other elements are not required to be added, so that not onlyis the safety higher, but also more simplicity can be realized; the synchronous expression of a plurality of guide RNAs with the Cas9 or the Cpf1 is realized by utilizing the one promoter, so that anexisting CRISPR editing system is simplified, moreover, the efficiency and the ability of simultaneously editing multiple target points of the CRISPR editing system are improved.

In accordance with the invention, novel gene deletions and translocations involving chromosome 2 resulting in fusion proteins combining part of Anaplastic Lymphoma Kinase (ALK) kinase with part of a secondary protein have now been identified in human solid tumors, e.g. non-small cell lung carcinoma (NSCLC). Secondary proteins include Echinoderm Microtubule-Associated Protein-Like 4 (EML-4) and TRK-Fusion Gene (TFG). The EML4-ALK fusion protein, which retains ALK tyrosine kinase activity, was confirmed to drive the proliferation and survival of NSCLC characterized by this mutation. The invention therefore provides, in part, isolated polynucleotides and vectors encoding the disclosed mutant ALK kinase polypeptides, probes for detecting it, isolated mutant polypeptides, recombinant polypeptides, and reagents for detecting the fusion and truncated polypeptides. The disclosed identification of this new fusion protein enables new methods for determining the presence of these mutant ALK kinase polypeptides in a biological sample, methods for screening for compounds that inhibit the proteins, and methods for inhibiting the progression of a cancer characterized by the mutant polynucleotides or polypeptides, which are also provided by the invention.

The invention relates to escherichia coli-derived single-plasmid-tandem soluble coexpression foot-and-mouth disease virus capsid proteins VP0 (which is a VP4 and VP2 fusion gene), VP1 and VP3, and a foot-and-mouth disease virus capsid protein virus-like particle preparation method. Foot-and-mouth disease virus capsid protein virus-like particles can be used for preparation of a foot-and-mouth disease vaccine. According to the method, a plurality of aspects of escherichia coli-derived soluble coexpression foot-and-mouth disease virus capsid protein are studied, by comprehensive use of tandem coexpression and SUMO(suggested upper merged ontology) technology with a tag for soluble coexpression of the foot-and-mouth disease virus capsid proteins VP0 (which is the VP4 and VP2 fusion gene), VP1 and VP3, the ultimate objective protein accounts for about 20% of total bacterial protein, and the foot-and-mouth disease virus capsid proteins obtained by purification can be successfully assembled into the virus like particles.

The invention relates to a method for biosynthesis preparation of human glucagon-like peptide-1 (GLP-1) and an analogue thereof. With adopting of a gene engineering technology, a recombinant escherichia coli expressed GLP-1 fusion protein is constructed, and a protein enzyme digestion site is designed in the fusion protein; a fusion gene has a gene sequence with a form of A-B-C structure, wherein A is a chaperonin gene, B is a nucleotide sequence encoding a connection peptide containing the enzyme digestion site, and C is a gene encoding the GLP-1 or the analogue thereof. After recombinant engineering bacteria is subjected to induced expression, the fusion protein is purified and subjected to enzyme digestion, and then the GLP-1 and the analogue thereof are obtained and are detected to have biological activity. The preparation method of the GLP-1 and the analogue thereof provided by the invention is simple and quick, the production conditions are mild, the product is convenient to separate and extract, the process is simple, and the industrialization prospect is good.

The invention provides a preparing method of reconstructing immunoregulatory protein of a ganoderma lucidum. The method includes the following steps: construction of engineering bacteria, wherein, LZ-8 protein gene is designed and coded, and is connected with a leading peptide coding sequence of saccharomyces cervisiae alpha-factor into fusion gene, which is transferred to a Pichia Pastoris expression system GS115 for reconstructing expression, high expressed rLZ-8 protein bacterial strain is acquired; the fermentation under the scale of 100L fermentation cylinder, wherein, improved seed culture medium and fermentation medium are used, additions of glycerol and methanol are regulated, 0.5M(NH4)2SO4 is added at the late stage of the fermentation; the purification on a column via centrifugal separation, wherein, salt is hyperfiltrated, powder is prepared after being frozen and dried. The method has large scale of fermentation, optimized technological condition and high production rate.

The present invention discloses primers, probes, a detection system and a kit for one time detection of lung cancer multiple genes, wherein the primers, the probes, and the distribution way for detecting 25 EGFR gene mutations, 7 KRAS gene mutations, 6 BRAF gene mutations, 9 NRAS gene mutations, 5 HER2 gene mutations, 2 PIK3CA gene mutations, 5 fusion genes of ALK5, 13 fusion genes of ROS1, and 6 fusion genes of RET are provided. According to the present invention, the detection kit adopts the 12 linking PCR reaction strip design, each 12 linking PCR strip detects multiple genes of a sample, and the corresponding fusion detection reagents and the internal control reagents are filled in the pipes 1-4 of the 12 linking PCR strip; and with the primers, the probes, the detection system and the kit, the one-time detection of the 24 fusions and the 54 mutations of the lung cancer can be achieved, such that the detection time is substantially shortened, the sensitivity is high, the specificity is strong, the operation is simple and rapid, and the reference for selection of tumor targeting drug therapy on lung cancer patients can be provided for clinician.

Disclosed herein is the discovery of a novel hepatitis C virus (HCV) isolated from a human patient. Embodiments of the inevntion include HCV peptides, nucleic acids encoding said HCV peptides, antibodies directed to said peptides, compositions containing said nucleic acids and peptides, as well as, methods of making and using the aforementioned compositions including, but not limited to, diagnostics and medicaments for the treatment and prevention of HCV infection.

The invention relates to detection of genotypes, in particular to a primer and a probe for detecting relative fusion genes of leukemia and a kit of the primer and the probe. The kit can qualitatively detect quantity of BCR-ABL, AML1-ETO and PML-RARa of the relative fusion genes of the leukemia in human marrow specimens so as to instruct clinical diagnosis and pharmacy of the leukemia.

The invention relates to a pathogenic microorganism genome database and an establishment method thereof, and belongs to the technical field of meta-genomes. The method comprises the following steps ofdata acquisition, wherein pathogenic microorganism genome data is obtained; strain genome screening, wherein species strain genomes are selected according to a predetermined screening rule; plasmid sequence removal, wherein plasmid sequences existing in the strain genomes obtained in the last step are removed; filtration, wherein according to a predetermined filtering rule, strains with incorrectlabeling information, incomplete chromosome assembly and incorrect classification are removed to obtain a reference strain genome of the species; fusion genome construction, wherein the reference strain genome is interrupted, redundancy is removed, reassembly is performed, and the sequences are reassembled to obtain a fusion genome of the species; database assembly, wherein the above steps are repeated to obtain the fusion genome of the predetermined species, summary is performed, and the pathogenic microorganism genome database is obtained. The genome database has the advantages of not onlyhaving a high precision rate, but also having short analysis time and reducing the cost.

The invention discloses a method and a device for detecting gene fusion, wherein the method comprises the following steps: S1, extracting RNA of a to-be-detected sample, and performing reverse transcription on RNA to obtain cDNA; S2, designing primers on two sides of a breaking point of a known fusion gene, and taking cDNA as a template for amplifying to form a sequence with a fusion form, and establishing a sequencing library; S3, sequencing the sequencing library by a high-throughput method to obtain a sequence with a fusion form; S4, detecting the sequence with the fusion form, particularly, S41, using comparison software to compare the sequence with the fusion form, obtained from the step 3, with a corresponding reference sequence with the fusion form; S42, judging whether data obtained from the step S41 meet the analysis requirements or not; S43, detecting the known fusion forms. The method has the advantages of being high in sensitivity and specificity and relatively economic.

The invention discloses a transcriptome-based tumor antigen identification method. The method comprises four steps of: obtaining an RNA sample of a patient tumor tissue, and carrying out library construction and amplification on the RNA sample to obtain an RNA sample sequencing result of the tumor tissue; aligning short read segments of the RNA sample sequencing result to a human reference genometo obtain an RNA alignment result; calculating gene expression quantity according to the RNA alignment result, and carrying out mutation detection and prediction of fusion gene events according to theRNA alignment result; and predicting transcriptome HLA typing according to the alignment result, wherein calculation of the gene expression quantity, mutation detection and prediction of the fusion gene events are carried out according to a specified order or simultaneously carried out; and using the gene expression quantity of a transcriptome sample, depth of transcriptome mutation sites in a whole-exon sequencing sample and binding force of neonatal short peptides and the patient HLA typing as an analysis result to submit the same to a downstream analyst. The invention provides the method capable of identifying a tumor-specific antigen of an individual sample from tumor patient transcriptome NGS data.

The invention relates to an EML4-ALK (echinoderm microtubule associated protein like4-anaplastic lymphoma kinase) fusion gene fluorescent quantitative PCR (polymerase chain reaction) assay kit, which is applicable to assay of EML4-ALK fusion gene mutation in lung adenocarcinoma. The kit comprises probes, primers and positive controls, which are specially designed for conserved sequences of 9 fusion variations of the EML4-ALK fusion gene. The kit can be used for quickly and accurately assaying 9 most common EML4-ALK fusion gene variations with high sensitivity, namely 9 fusion variations of 7 variant subtypes, which are subtype 1 (E13; A20), subtype 2 (E20; A20), subtype 3 (E6a/b; A20), subtype 4 (E14; A20), subtype 5 (E2a/b; A20), subtype 6 (E18; A20) and subtype 7 (E14; A20), so that a real-time fluorescent quantitative PCR system for assaying 9 most common EML4-ALK fusion gene variations can be established.

The invention provides a method for detecting the off-target effect of an adenine base editor system based on whole genome sequencing, and its application in gene editing, The adenine base editor system comprises a TadA:TadA*:Cas9 fusion gene and gRNA. The system can catalyze the efficient replacement of adenine (A) at a target site with guanine (G), and has a broad application prospect in human disease gene editing therapy and disease model construction. The first method for detecting the off-target effect of the ABE system in a whole genome range, called EndoV-seq method for short, is developed in the invention. The EndoV-seq method provided by the invention has a broad application prospect in the field of gene editing, especially gene editing therapy.

The invention belongs to the technical field of biological engineering, and relates to an NGF-Fc fusion protein and a preparation method thereof. The method comprises the following steps: constructing a human immune globulin IgG Fc and nerve growth factor (NGF) fusion gene expression vector through a genetic engineering means, transferring to a mammal cell to make the transferred mammal cell highly express and produce NGF-Fc fusion proteins, purifying, identifying and carrying out biological activity detection. The expression vector transferred mammal cell constructed in the invention can express bioactive NGF-Fc fusion proteins, so the expression level can be 150mg/L or above, and the obtained protein has good stability and long half life, and can be used to prepare drugs for treating Alzheimer's disease, diabetic peripheral neuropathy, Parkinson's disease, facial neuritis, craniocerebral trauma, trauma of spinal cord, acute cerebrovascular disease, encephalatrophy and other neurological diseases, and peripheral nerve injury acute cerebral vascular central nerve injuries induced by chemical drugs.

The invention discloses a leukemia fusion gene combined parallel detecting method and a diagnostic reagent kit. The combined parallel detecting method is characterized in that multiple fusion genes related to leukemia can be detected at one time. By designing a primer and a probe of the leukemia fusion gene mRNA, the mixture covalently combined by the probe and microspheres is hybridized with a reverse transcription PCR amplified product, the streptavidin of phycoerythrin is added, and then the fluorescence signals of different microspheres can be detected, thereby determining whether the sample to be detected contains leukemia fusion genes or not and the express conditions of the fusion genes. The diagnostic reagent kit has the advantages of high sensitivity, high flux property, quick and accurate detection, and the like, and can simultaneously carry out qualitative and quantitative detection on multiple leukemia fusion genes.

In accordance with the invention, novel gene deletions and translocations involving chromosome 2 resulting in fusion proteins combining part of Anaplastic Lymphoma Kinase (ALK) kinase with part of a secondary protein have now been identified in human solid tumors, e.g. non-small cell lung carcinoma (NSCLC). Secondary proteins include Echinoderm Microtubule- Associated Protein-Like 4 (EML-4) and TRK- Fusion Gene (TFG). The EML4-ALK fusion protein, which retains ALK tyrosine kinase activity, was confirmed to drive the proliferation and survival of NSCLC characterized by this mutation. The invention therefore provides, in part, isolated polynucleotides and vectors encoding the disclosed mutant ALK kinase polypeptides, probes for detecting it, isolated mutant polypeptides, recombinant polypeptides, and reagents for detecting the fusion and truncated polypeptides. The disclosed identification of this new fusion protein enables new methods for determining the presence of these mutant ALK kinase polypeptides in a biological sample, methods for screening for compounds that inhibit the proteins, and methods for inhibiting the progression of a cancer characterized by the mutant polynucleotides or polypeptides, which are also provided by the invention.

The invention discloses a human ALK fusion gene detection primer set and a detection kit. The primer set comprises one or more detection primers in sixteen ALK fusion gene types A01 to A16 and Taqman probes corresponding to the gene types. As many as sixteen fusion gene types except for EML4 can be detected through the detection kit formed by the primer set and the probes, the requirement for the sample RNA obtaining quality is low, and the ideal detection effect can be achieved through samples in different existence forms; by means of the kit, an operator only needs to directly feed extracted sample RNA once, inverse transcription and PCR amplification are carried out in one closed tube reaction system, and the pollution probability and the result error probability are reduced. One-time detection only takes eighty minutes. The human ALK fusion gene detection primer set and the detection kit have the remarkable advantages that the number of the detection types is large, sensitivity is high, experiment operation is simple, the period is short, the human ALK fusion gene detection primer set and the detection kit are safe and nontoxic, and cost is low.