119 results about "Gene defect" patented technology

The invention discloses a CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 system and application thereof to construction of swine-derived recombinant cells with insulin receptor substrate gene defects. The invention provides an sgRNA (ribonucleic acid) combination which is composed of sgRNAIRS1-1, sgRNAIRS1-3, sgRNAIRS2-2 and sgRNAIRS2-3. The invention provides a plasmid combination. The plasmid combination is composed of four plasmids, and the four plasmids are respectively subjected to transcription to obtain sgRNAIRS1-1, sgRNAIRS1-3, sgRNAIRS2-2 and sgRNAIRS2-3. Theapplication of the sgRNA combination or the plasmid combination is as follows: preparation of recombinant cells; preparation of a diabetic cell model; preparation of a diabetic animal model. Accordingto the invention, sgRNAIRS1-1 is as shown in SEQ ID NO:8; sgRNAIRS1-3 is as shown in SEQ ID NO:10; sgRNAIRS2-2 is shown as SEQ ID NO:14; and sgRNAIRS2-3 is as shown in SEQ ID NO:15. A solid foundation is laid for the preparation of diabetic pig models, and important application value for the research and development of diabetic medicaments is achieved.

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 present invention discloses a construction of three-gene defected recombinant pseudorabies virus (PrV) strain, vaccine prepared by said strain, method for constructing said strain and method for preparing said vaccine. The described recombinant pseudorabies virus strain defects genes of TK.gE ang gI, and contains no exogenous gene. The described vacine belongs to the freeze-dried live vaccine made up by virus liquid containing said invention and gelatin. Said invented live vaccine can be inoculated on the piglet, slaughter pig and sow with farrow, and can obtain obious immune effect. Said vaccine has good biological safety, and can be used for preventing and curing pseudorabies.

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 provides construction of a lactobacillus acidophilus S-layer protein surface display system, which belongs to the field of biotechnology. The construction is characterized in that gene acquisition comprises (1) PCR amplification, (2) the connection of a SlpA gene and a pMD18-TVector system and (3) the identification of cloning plasmid pMD18T-S, and the construction comprises (1) theacquisition of a target gene and a double-labeling expression vector, (2) the connection and transformation of the double-labeling expression vector and SlpA, (3) the identification of a surface display vector system and (4) the detection of the anchored expression situation of S-layer protein on recombinant surface. The construction has the advantages that: 1, the constructed vector can be used to be transformed into S-layer protein gene defect lactobacillus to study the formation mechanism of lactobacillus S-layer protein; and 2, by utilizing a DNA recombination technique, protective antigengenes of pathogens are fused with lactobacillus S-layer protein on the surface display vector.

The invention relates to the technical field of animal models and concretely, relates to a method for building an animal model with PHF 14 gene knockout-based acute kidney injury and renal fibrosis after injury. The invention builds a model of renal fibrosis after kidney injury under the genetic background of PHF 14 gene knockout under adult conditions and provides essential conditions for researching a use of a PHF 14 gene in fibrotic diseases. Compared with the existing PHF 14 knockout model, the PHF 14 gene knockout provided by the invention does not cause embryonic lethal. A test on adult-stage tamoxifen induced knockout of PHF 14 proves that the PHF 14 gene knockout does not cause embryonic lethal. Compared with a group without knockout, the group subjected to knockout has no substantial pathology of lung, liver and kidney. The method prevents systemic disease state-caused interference on following-up acute kidney injury model making and realizes individual research on use of the gene defect in renal fibrosis pathogenesis.

The invention belongs to the technical field of molecular biology and in particular relates to a preparation method of a PD-1 (Programmed cell death protein 1) and CTLA4 (Cytotoxic T-Lymphocyte Antigen 4) double-gene defect type T lymphocyte preparation. Construction of a PD-1 and CTLA4 double-gene knockout vector, T lymphocyte separation and activation, electric transfection of T lymphocytes and T7E1 enzyme digestion identification, and flow cytometer screening and sequencing analysis are carried out. According to the production of a gene defect type immune cell preparation, on one hand, a production procedure of cells is simplified; on the other hand, the capability of killing tumor cells by the T lymphocytes is improved and the long-period tumor immunity can be enhanced.

The invention discloses a detection kit for Thrombophilia-related gene mutation. The detection kit comprises ddH2O, 10*buffer (Takara), 2*GC buffer I, dNTP, MgCl2, HSTaq, multiplex PCR primer mixed liquid, Illumina I5/I7 primer series reagents. The detection kit has the advantages that the detection kit uses the multiplex PCR enriching next generation sequencing technology to comprehensively and systematically detect the point mutation of all exons and control regions of 18 genes which include PROC, PROS1, THBD, PROCR, F5, AT3, HCF2, F2, F7, F8, F9, F10, F11, F12, PLG, ADAMTSl3, HRG and TFPI and performs detection aiming at the specific polymorphic sites of three genes which include PLAT, FGG and HABP2; the detection kit is used for biological researches and molecularly diagnosing the Thrombophilia caused by the defects of the genes above; the detection kit is wide in detection range, high in throughput, fast and accurate in detection method, high in detection accuracy, low in cost, and the like.

The invention discloses a gluconobacter oxydans nonresistant marker gene knockout system, and belongs to the technical field of genetic engineering. According to the invention, through a homologous recombination technique, upp genes in G.o xydans are knocked out, so that a upp genetic deficient strain is obtained; a recombinant integrating plasmid containing a upp gene which can be subjected to good transcribed expression in G.o xydans and has a conditional lethal function, a resistance maker kanamycin resistant marker gene for the transformant selection of G.o xydans and an MCS polycloning site is built. The plasmid is applicable to the knockout and replacement of G.o xydans chromosome genes; compared with gene knockout implemented by taking a resistant marker gene as a selection marker, the system disclosed by the invention has the following two advantages: no resistant gene is introduced to a G.o xydans genome, so that a polar effect of the resistant gene on upstream and downstream genes is avoided; by using the knockout system, the multiple knockout and replacement of chromosome genes can be performed in a same strain.

The invention relates to the field of biology, and provides a genetics detecting device of an embryo before implantation. The genetics detecting device of the embryo before implantation comprises a message unit 100, an amphiphilic haplotype creating module 200, a candidate embryo haplotype building module 300, a candidate embryo haplotype parental source determining module 400 and a judgment module 500 for judging whether a candidate embryo carries gene defects. According to the genetics detecting device of the embryo before implantation provided by the invention, a more accurate genetics diagnosis of the embryo before implantation can be given, so as to select out healthy embryos.

The invention discloses a function and application of SH2B adapter protein 3(SH2B3) for treating cardiac hypertrophy. By virtue of the SH2B adapter protein 3(SH2B3), interrelation between the expression of SH2B3 gene and cardiac hypertrophy is determined; when the cardiac hypertrophy occurs, the expression of SH2B3 is obviously increased; activation of an Akt signal channel is suppressed by SH2B3 gene defect, the cardiac hypertrophy and fibrosis are also suppressed, and the heart function is protected; the activation of the Akt signal channel is remarkably accelerated by over expression of the SH2B3 gene, the cardiac hypertrophy and fibrosis are improved, and the heart function is deteriorated. Therefore, the SH2B3 gene can be used as a medicine target mark for screening medicines for protecting the heart function and/or preventing, relieving and/or treating the cardiac hypertrophy; the SH2B3 gene can be used as a target gene in gene treatment for designing and preparing medicines and/or biological reagents for protecting the heart function and/or preventing, relieving and/or treating the cardiac hypertrophy, so that an effective novel way is provided for the treatment of the cardiac hypertrophy.

The invention relates to a gene NS1 (Structural Protein) defective respiratory syncytial virus and application thereof. The gene NS1 defective respiratory syncytial virus is synthesized by Vero cells, and the Vero cells are formed through plasmid cotransfection of genes carrying NS1-defective virus cDNA, viruses N, M2-1, P and L. In the gene NS1 defective respiratory syncytial virus, the gene NS1 in the respiratory syncytial virus is deleted by utilizing a reverse gene strategy. The gene NS1 defective respiratory syncytial virus is applied to treating breast cancer and kills breast cancer cells selectively.

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 gene aroAS001 for encoding 5-enolpyruvyl-shikimate-3-phosphate synthase, and an application of the gene, wherein the base sequence of the gene is represented by SEQ ID No.3; the gene is suitable for enhancing glyphosate resistance of crops; specifically, gene aroAS001, and an expression vector or a cloning vector containing gene aroAS001, or strains containing the expression vector or cloning vector are transferred in the crops. The gene provided by the invention is an excellent glyphosate resistant gene, the aroA gene defect type strains transferred with the gene can grow in a culture medium containing lower than 400mM glyphosate; compared with other known EPSP (enolpyruvyl-shikimate-3-phosphate) synthase genes, the gene provided by the invention has significant glyphosate resistance and brings about unexpected beneficial effects.

The invention discloses a function and application of SH2B adapter protein 1 (SH2B1) in myocardial hypertrophy treatment. According to the function and application, the mutual relation between SH2B1 genetic expression and myocardial hypertrophy is determined; the SH2B1 expression rises obviously when the myocardial hypertrophy happens; SH2B1 interference and overexpression adenovirus are used for restraining and promoting cardiomyocyte hypertrophy respectively; the SH2B1 gene defect and overexpression are used for restraining and obviously raising Jak2/Stat3 signal pathway activation respectively, so that cardiac hypertrophy and fibrosis are restrained and promoted, and the cardiac function can be protected and deteriorated. Hence, the SH2B1 gene can be used as a drug target to be used for screening drugs for protecting the cardiac function and/or preventing, relieving and treating cardiac hypertrophy; the SH2B1 gene can be used as a target gene in gene therapy to be used for designing and preparing the drugs for protecting the cardiac function and/or preventing, relieving and treating cardiac hypertrophy; a new path is provided for cardiac hypertrophy treatment.

The invention provides a single-gene defect detection method, a primer composition for single-gene defect detection in a Xq28 region and a kit. The method comprises the following steps: designing the primer composition; acquiring DNA (deoxyribonucleic acid) samples of a proband, parents and offspring; carrying out amplification by using the primer composition, constructing a library and carrying out sequencing analysis; and obtaining haplotypes of the proband, the parents and the offspring based on a sequencing analysis result, and determining a detection result of the single-gene defect of the offspring. The design method of the primer composition comprises the following steps: acquiring SNP loci of a to-be-detected DNA fragment, screening SNP detection loci and designing primer pairs; or obtaining the SNP loci and the STR loci of the to-be-detected DNA fragment, screening the SNP detection loci and the STR detection loci, and designing the primer pairs. The method, the primer composition and the kit provided by the invention can be used for identifying whether the gene in the to-be-detected DNA fragment of an embryo before implantation varies or not, and the qualified embryo is efficiently and accurately screened for implantation, so that the disease risk is reduced.

The invention provides a method for establishing a heart disease drug screening model, which comprises the following steps of A) establishing a virus vector comprising four transcription factors: Oct3/4, Sox2, c-Myc and klf4; B) transfecting the virus vector in the step A) with a virus packaging cell to obtain virus liquid; C) infecting the virus liquid in the step B) with the fibroblast of the beta adrenergic receptor wild-type gene knockout mouse; D) screening the clone similar to the mouse embryonic stem cell, and subculturing to obtain the induced pluripotent stem cells with beta adrenergic receptor wild-type gene defect; E) establishing a virus vector containing the beta adrenergic receptor mutant gene; F) transfecting the virus vector in the step E) with the virus packaging cell to obtain virus liquid; G) infecting the virus liquid in the step F) with the induced pluripotent stem cells with beta adrenergic receptor wild-type gene defect; H) selecting positive clone and performing induced differentiation into myocardial cells.