122results about How to "Increased transcript levels" patented technology

The invention discloses an anti-BCMA chimeric antigen receptor, an encoding gene, a recombinant expression vector and an establishing method and application of the anti-BCMA chimeric antigen receptor, the encoding gene and the recombinant expression vector. The receptor comprises a CD8 leader chimeric receptor signal peptide, a BCMA single-chain antibody heavy chain VH, an Optimal Linker C, a BCMA single-chain antibody light chain VL, a CD8 Hinge chimeric receptor hinge, a CD8 Transmembrane chimeric receptor transmembrane domain, a CD137 chimeric receptor co-stimulatory factor and a TCR chimeric receptor T cell activating domain which are sequentially connected in series. In addition, the invention further discloses the encoding gene and the recombinant expression vector of the anti-BCMA chimeric antigen receptor and the establishing method and application of the encoding gene and the recombinant expression vector. The secretion of cell factors and the cytotoxicity in vitro of CAR-T cells can be remarkably improved, and the clinical treatment effect is outstanding.

The invention provides a cotton promoter GbU6-7PS and application. The nucleotide sequence of the promoter is as shown in SEQ ID NO.1, and the nucleotide sequence of a complementary strand of the promoter is as shown in SEQ ID NO.2. The cotton promoter GbU6-7PS is obtained by conducting the first turn of PCR amplification so as to obtain a segment which covers full-length GbU6-7 promoter sequence, so that GbU6-7P is obtained; and then, conducting the second turn of PCR amplification, taking the GbU6-7P as a donor plasmid template and GbU6-5P::GUS as an acceptor plasmid template, and truncating and cloning the GbU6-7 promoter by virtue of Transfer PCR method, so that the cotton promoter GbU6-7PS is obtained. The cotton promoter GbU6-7PS disclosed by the invention is not only applicable to cotton, but also quite short in segment which is just 190bp long, conforming to the requirement of a CRISPR / Cas9 genome editing vector, the transcriptional level of sgRNA is significantly improved, and subsequently a genome editing efficiency may be increased.

The invention provides a method for obtaining pancreatic precursor cells and pancreatic beta cells through differentiation of human multipotential stem cells. In the process of specializing entoderm cells derived from the human multipotential stem cells towards pancreas pedigree cells, the efficiency of differentiating the human multipotential stem cells towards the pancreatic precursor cells canbe improved by stepwise adding a WNT signal channel inhibitor. According to the method, after the efficiency of differentiation towards the pancreatic precursor cells is improved, the efficiency of differentiation of the mature pancreatic beta cells is improved accordingly. The method is suitable for pancreas differentiation of different cell lines, a large quantity of pancreatic beta cells are obtained, strong support is provided for diabetes cell therapy, drug screening, disease models and the like, and the method has a good application prospect.

The invention discloses an acetyl coenzyme A acetyltransferase gene RKAcaT2 and application thereof. The nucleotide sequence of the gene is shown in SEQ ID NO:1, the amino acid sequence encoded by thegene is shown in SEQ ID NO:2, the gene is a key enzyme gene synthesized by carotenoid in Rhodosporidium kratochvilovae YM25235 and has the functions of the acetyl coenzyme A acetyltransferase, and the carotenoid produced by the Rhodosporidium kratochvilovae YM25235 can be controlled; microorganisms are transformed through the genetic engineering means so as to increase the yield of carotenoid inthe microorganisms, and a foundation is laid for large-scale commercial production of the carotenoid.

The invention discloses an anti-CD20 chimeric antigen receptor, an encoding gene, a recombinant expression vector, a construction method of the recombinant expression vector and an application. The anti-CD20 chimeric antigen receptor comprises CD8 leader chimeric receptor signal peptide, CD20 VL, Optimal Linker C, a CD20 single-chain antibody heavy chain VH, a CD8 Hinge chimeric receptor hinge, a CD8 Transmembrane chimeric receptor transmembrane domain, a CD137 chimeric receptor co-stimulated factor and a TCR chimeric receptor T cell activating domain which are connected in series. Moreover, the invention discloses an encoding gene of the anti-CD20 chimeric antigen receptor, a recombinant expression vector, a construction method of the recombinant expression vector, and an application. Secretion of cell factors and an in-vitro killing effect of CAR-T cells are obviously improved, and the CAR-T cells have an outstanding clinical treatment effect.

The invention discloses an anti-EGFRvIII chimeric antigen receptor, an encoding gene, a recombinant expression vector, a construction method of the recombinant expression vector, and an application. The anti-EGFRvIII chimeric antigen receptor comprises CD8 leader chimeric receptor signal peptide, an EGFRvIII single-chain antibody light chain VL, Optimal Linker C, an EGFRvIII single-chain antibody heavy chain VH, a CD8 Hinge chimeric receptor hinge, a CD8 Transmembrane chimeric receptor transmembrane domain, a CD137 chimeric receptor co-stimulated factor and a TCR chimeric receptor T cell activating domain which are connected in series. Moreover, the invention discloses an encoding gene of the anti-EGFRvIII chimeric antigen receptor, a recombinant expression vector, a construction method of the recombinant expression vector, and an application. Secretion of cell factors and an in-vitro killing effect of CAR-T cells are obviously improved, and the CAR-T cells have an outstanding clinical treatment effect.

The invention discloses a phytoene dehydrogenase gene RKcrtI having the nucleotide sequence shown in SEQ ID NO:1; the amino acid sequence encoded by the gene is shown in SEQ ID NO:2. The gene is derived from rhodosporidium kratochvilovae YM25235; the gene is connected with a vector and then is transferred into yeast cells. The rhodosporidium kratochvilovae YM25235 can be promoted to produce beta-carotenoid, and a foundation is laid for large-scale commercial production of beta-carotenoid.

The invention discloses an anti-HER2 chimeric antigen receptor, an encoding gene, a recombinant expression vector, a construction method of the recombinant expression vector, and an application. The anti-HER2 chimeric antigen receptor comprises CD8 leader chimeric receptor signal peptide, an HER2 single-chain antibody heavy chain VH, Optimal Linker C, an HER2 single-chain antibody light chain VL, a CD8 Hinge chimeric receptor hinge, a CD8 Transmembrane chimeric receptor transmembrane domain, a CD137 chimeric receptor co-stimulated factor and a TCR chimeric receptor T cell activating domain which are connected in series. Moreover, the invention discloses an encoding gene of the anti-HER2 chimeric antigen receptor, a recombinant expression vector, a construction method of the recombinant expression vector, and an application. Secretion of cell factors and an in-vitro killing effect of CAR-T cells are obviously improved, and the CAR-T cells have an outstanding clinical treatment effect.

The invention discloses an HMG-CoA synthetase gene RKHMGCS, the nucleotide sequence is shown as the SEQ ID NO:1, and the amino acid sequence of the gene coding is shown as the SEQ ID NO:2; the gene isthe key enzyme gene for synthesis of the carotenoid in the Rhodosporidium kratochvilovae YM 25235, the gene has the HMG-CoA synthetase function, and the Rhodosporidium kratochvilovae YM 25235 can becontrolled to produce the carotenoid; the microorganism is modified by the means of genetic engineering, the yield of the carotenoid in the microorganism body is improved, and a foundation is laid forlarge-scale commercialized production of the carotenoid.

The invention discloses an anti-CD33 chimeric antigen receptor, a coding gene, a recombinant expression vector and a construction method and an application of the recombinant expression vector. The anti-CD33 chimeric antigen receptor comprises a CD8leader chimeric receptor signal peptide, a heavy chain VL of a CD33 single-chain antibody, an Optimal Linker C, a light chain VH of the CD33 single-chain antibody, a CD8Hinge chimeric receptor hinge, a CD8Transmembrane chimeric receptor transmembrane domain, a CD137 chimeric receptor co-stimulation factor and a TCR chimeric receptor T-cell activation domain, which are serially connected in sequence. In addition, the invention discloses the coding gene of the anti-CD33 chimeric antigen receptor, the recombinant expression vector and the construction method and the application of the recombinant expression vector. According to the anti-CD33 chimeric antigen receptor, the secretion of cell factors and the in vitro killing effect of CAR-T (Chimeric Antigen Receptor T-Cell Immunotherapy) cells can be remarkably improved, and the clinical treatment effect is outstanding.

The invention discloses a streptomycete constitutive promoter and applications thereof, and provides an DNA fragment. The DNA fragment is any one selected from the following DNA molecules in 1)-3): 1) an DNA molecule represented by the sequence 1 in a sequence table; 2) an DNA molecule with promoter functions and hybrid with the DNA molecule shown in 1) under strict conditions; 3) an DNA molecule with more than 90% of homology with the DNA molecule shown in 1) and with promoter functions. The experiments shows that a strong constitutive promoter kasO*p is screened out, and the promoter and the erythromycin promoter ermE*p are compared in representation of transcriptional level and expression level in streptomyces coelicolor, streptomyces avermitilis and streptomyces venezuelae, and the results show that the modified promoter kasO*p is better than the the erythromycin promoter ermE*p.

The invention discloses an acyl coenzyme A oxidase 2 gene RKACOX2, the nucleotide sequence of the acyl coenzyme A oxidase 2 gene RKACOX2 is shown as SEQ ID NO: 1, and the amino acid sequence coded bythe acyl coenzyme A oxidase 2 gene RKACOX2 is shown as SEQ ID NO: 2. The gene is separated from Rhodosporidium kratochvilovae YM25235, the gene is transferred into the Rhodosporidium kratochvilovae YM25235, experimental results show that overexpression of the RKACOX2 gene can cause improvement of the transcription level of the gene in cells to a certain extent, and overexpression of the RKACOX2 gene can promote synthesis of all components of total carotenoid and carotenoid. Microorganisms are modified through a genetic engineering means, the yield of microorganism carotenoid can be increased,and a foundation is laid for large-scale commercialized production of the carotenoid.

The invention discloses cloning and function-expressing methods of a peanut adversity stress AhROLP1 gene. The adversity stress AhROLP1 gene is synthesized by preparing and processing materials, extracting RNA (Ribonucleic Acid), synthesizing cDNA, and cloning with RT-PCR (Reverse Transcription-Polymerase Chain Reaction). An open reading frame of the gene is 1620 bp, and 540 amino acids in total are coded. An amino acid sequence of the gene has homology of over 70% with reticuline oxidase of plants such as chickpeas, soybeans, wild strawberries, grapes and tomatoes. An expression mode of the AhROLP1 under low temperature, salt stress and drought stress is verified through fluorescent quantitative PCR (Polymerase Chain Reaction), and the result indicates that transcriptional level of the gene is obviously increased under adversity stress of three non-living things, and then is kept at a higher level all the time. The gene is lead into arabidopsis through transgenic means, and a transgenic plant has obvious cold resistance, salt resistance and drought resistance in comparison with a control group.

The invention discloses an anti-CD138 chimeric antigen receptor, a coding gene, a recombinant expression vector and a construction method and an application of the recombinant expression vector. The anti-CD138 chimeric antigen receptor comprises a CD8leader chimeric receptor signal peptide, a light chain VH of the CD138 single-chain antibody, an Optimal Linker C, a heavy chain VL of a CD138 single-chain antibody, a CD8Hinge chimeric receptor hinge, a CD8Transmembrane chimeric receptor transmembrane domain, a CD137 chimeric receptor co-stimulation factor and a TCR chimeric receptor T-cell activation domain, which are serially connected in sequence. In addition, the invention discloses the coding gene of the anti-CD138 chimeric antigen receptor, the recombinant expression vector and the construction method and the application of the recombinant expression vector. According to the anti-CD138 chimeric antigen receptor, the secretion of cell factors and the in vitro killing effect of CAR-T (Chimeric Antigen Receptor T-Cell Immunotherapy) cells can be remarkably improved, and the clinical treatment effect is outstanding.

The present invention provides a composition for treating and / or preventing type I diabetes and an application thereof. The active ingredient of the composition is 1.) or 2.) or 3.), as follows: 1.) a mixture of a type I diabetes protein antigen and an immunosuppressor, 2.) a mixture of a type I diabetes protein antigenic epitope polypeptide and an immunosuppressor, 3.) a mixture of a type I diabetes protein antigen, a type I diabetes protein antigenic epitope polypeptide, and an immunosuppressor; the type I diabetes protein antigen is at least one of insulin, glutamic acid decarboxylase, and islet amyloid polypeptide, and the immunosuppressor is at least one of dexamethasone, cyclosporine A, tacrolimus, mycophenolate mofetil, azathioprine, prednisone, early prednisolone, anti-CD4 monoclonal antibody, and anti-CD3 monoclonal antibody.

The invention provides a set of antisense RNA for inhibiting AMH gene expression and a method for promoting gonad degeneration of male tilapia mossambica and increasing weight gain, and belongs to thetechnical field of aquaculture breeding. The antisense RNA for inhibiting AMH gene expression comprises antisense RNA1 and antisense RNA2, can accurately inhibit the transcriptional level of anti-mullerian hormone gene mRNA, interferes mRNA transport and translation of the anti-mullerian hormone gene mRNA, and influences the protein expression level of the anti-mullerian hormone gene mRNA. The invention further provides the method for promoting gonad degeneration of male tilapia mossambica and increasing weight gain. Two antisense RNAs are introduced into ova by adopting an antisense RNA sequence introduction technology, damage to the ova is small, and the success rate is high. The method can effectively carry out target gene research and development of new disease-resistant and growth-promoting varieties, and has a very strong application prospect.

The invention discloses an acetyl CoA synthetase gene RKACS1, the nucleotide sequence of which is as shown in SEQ ID NO: 1, and the amino acid sequence encoded by the gene is as shown in SEQ ID NO: 2. The gene is the acetyl CoA synthetase gene separated from Rhodosporidium kratochvilovae YM25235, the gene is transformed into the Rhodosporidium kratochvilovae YM25235, the experimental result shows that the overexpression of the acetyl CoA synthetase gene RKACS1 can cause the increase of the total carotenoid content and the grease content in the strain of the Rhodosporidium kratochvilovae YM25235. Microorganisms are modified through a genetic engineering means, so that the yield of the carotenoid and the grease in microorganisms is improved, and a foundation is laid for large-scale commercial production of the carotenoid and the grease.

The invention relates to an acidithiobacillus caldus gene engineering strain and applications thereof. The acidithiobacillus caldus MTH-04(pJRD215-tac-sor) gene engineering strain is preserved in the general microbiology centre of China Committee for Culture Collection of Microorganisms on June 28, 2013, address: Institute of Microbiology Chinese Academy of Sciences, NO.3, NO.1 yard, beichen west road, Beijing chaoyang district. The strain preservation number: CGMCC NO.7388. Compared with the acidithiobacillus caldus MTH-04, the acidithiobacillus caldus MTH-04(pJRD215-tac-sor) gene engineering strain has the advantages that the capability of sulfur oxide is enhanced, and the bacteria biomass, sulfur and oxygen reductase enzyme activity and the transcriptional level of key enzyme of a sulfur oxidation system are increased.

The invention discloses a 3-ketoacyl-CoA thiolase gene RkACAA1-1. A nucleotide sequence of the 3-ketoacyl-CoA thiolase gene RkACAA1-1 is shown as SEQ ID NO: 1, and an amino acid sequence coded by the gene is shown as SEQ ID NO: 2. The gene is separated from rhodosporidium kratochvilovae YM25235, the gene is transformed into the rhodosporidium kratochvilovae YM25235, and an experimental result shows that the overexpression of the 3-ketoacyl-CoA thiolase gene RkACAA1-1 can cause the increase of total carotenoids in a rhodosporidium kratochvilovae YM25235 strain. According to the 3-ketoacyl-CoA thiolase gene RkACAA1-1 and the application thereof, the rhodosporidium kratochvilovae is modified by a genetic engineering means to increase the content of the carotenoids, so that good application prospects and economic benefits are provided for industrial production of the carotenoids, and a foundation is laid for large-scale commercial production of the carotenoids.

The invention discloses a cDNA sequence of ERF transcription factor from Halimodendron halodendron, as well as an expression vector and application thereof, which belongs to the field of plant genetic engineering. A novel ERF2 ERF2 transcription factor gene HhERF2 (SEQ ID NO:1) is obtained by separating and cloning psammophyte Halimodendron halodendron through a RACE-PCR technique. The result of tissue-specific expression shows that the gene is expressed in roots, stalks and leaves of the Halimodendron halodendron. The result of stress response detection shows that the expression of the gene is induced by droughty, high-salt and low-temperature environmental stress and does not depend on the regulation of ABA. The cDNA sequence is transformed into Arabidopsis for expression; the stress resistance of transgenic Arabidopsis to drought, high salt, low temperature, diseases and the like is analyzed; and the relevant physiological and biochemical indexes of the transgenic Arabidopsis are determined. Experimental result shows that the cDNA sequence can effectively improve the drought resistance, osmotic-stress resistance and disease resistance of plants.

The invention discloses a method for promoting the accumulation of flavonoid substances of ornamental Chinese flowering crabapple leaf tissues. The method comprises the following steps: by adopting the ornamental Chinese flowering crabapple leaf variety as a material, designing melatonin of different concentration gradients to treat an ornamental Chinese flowering crabapple tissue culture seedling, selecting the leaves treated by 200 micrograms / l of melatonin according to phenotype, wherein the leaf is apparent in color, the color of the leaf treated by the melatonin with higher concentrationor lower concentration is not apparently changed, then detecting a content of the flavonoid substances in the ornamental Chinese flowering crabapple leaves treated by the 200 micrograms / l of melatoninas well as a transcriptional level of a key gene in the flavonoid metabolism way, and a transcriptional level of the melatonin way related gene, wherein a result shows that in the ornamental Chineseflowering crabapple treated with the 200 micrograms / l of melatonin, the content of anthocyanin is the highest. A method is provided for improving the reversion phenomenon due to the increase of the subculture times of the ornamental Chinese flowering crabapple, and a good theoretical foundation is further provided for the genetic improvement of the color-leafed plant in the future.

The invention relates to a Leymus chinensis aquaporin and an encoding gene and application thereof. The current researches of Leymus chinensis are only to perform eating and composition analysis for human or livestock, but the researches in molecular biology aspect, regarding Leymus chinensis as drought tolerant crop are less. The Leymus chinensis aquaporin is the protein with the following amino acid sequences: (1) the amino acid sequence in SEQ ID No:1 of the sequence table; and (2) the protein derived from the protein with the amino acid sequence in SEQ ID No:1, with the same activity of the amino acid sequence in SEQ ID No:1 of the sequence table by replacing, deleting or adding one or more of amino acid residues to amino acid sequence in SEQ ID No:1 of the sequence table. The encoding gene of the Leymus chinensis aquaporin of the invention can be transferred in other plants to increase the drought-resisting property of transgenic plants. The Leymus chinensis aquaporin and encoding gene of the invention are used in plant gene engineering field.

The invention discloses a method for cloning AhbHLH1L genes and expressing functions for peanuts under adversity stress. The method is characterized by mainly comprising steps of (1), preparing and processing materials; (2), extracting RNA (ribonucleic acid) and synthesizing cDNA [complementary DNA (deoxyribonucleic acid)]; (3), cloning the genes. The method has the advantages that expression modes of the AhbHLH1L genes at low temperatures and under salt stress and drought stress are verified by means of fluorescence quantitative PCR (polymerase chain reaction), and the fact that the stress resistance of plants can be obviously improved by the AhbHLH1L genes is declared.

The invention discloses a genetically engineered bacterium for producing natamycin as well as a construction method and application of the genetically engineered bacterium. The genetically engineered bacterium is classified and named Streptomyces gilvosporeus and is collected with the serial number of CGMCC No.8901 in the China general microbiological culture collection center on march 7th, 2014. The construction method comprises the steps of constructing a recombinant plasmid by utilizing a cholesterol oxidase gene pimE; integrating the gene pimE into a Streptomyces gilvosporeus genome in an Escherichia coli and Streptomyces gilvosporeus conjugal transfer way; sieving to obtain a positive transconjugant. The genetically engineered bacterium can be used for increasing the density of pimE proteins in a fermentation liquid, and as signaling proteins, the pimE proteins can be used for improving the transcriptional level of a natamycin gene cluster and greatly increasing the fermentation yield of natamycin, so that the production cost is reduced, and huge social and economic benefits are brought.

The invention discloses a 3-ketoacyl coenzyme A thiolase gene RKACAA1-2, the nucleotide sequence of the 3-ketoacyl coenzyme A thiolase gene RKACAA1-2 is shown as SEQ ID NO: 1, and the amino acid sequence coded by the gene is shown as SEQ ID NO: 2; the gene is separated from rhodosporidium kratochvilovae YM25235, the gene is transferred into the rhodosporidium kratochvilovae YM25235 through transformation, and the experimental result shows that the overexpression of the RKACAA1-2 gene can cause the improvement of the carotenoid synthesis level of the YM25235 strain; according to the gene, the microorganisms are modified by means of genetic engineering, so that the yield of carotenoid in the microorganisms is increased, and a foundation is laid for large-scale commercial production of carotenoid.

The invention relates to the technical field of plant gene engineering, in particular to the separation, cloning and applications of a TtLRR-STK gene the full length cDNA of which is 3081bp. The TtLRR-STK gene is separated from the emmer with high resistance to wheat diseases; the gene codes the receptor protein kinase of a leucine-rich area-serine / threonine; and the receptor protein kinase includes a plurality of parts such as an N-end conserved sequence, a leucine-rich structure domain (LRR), a middle transmembrane structure domain, a C-end protein kinase structure, and the like. The structure thereof is similar to a cloned rice resistance to bacterial blight protein Xa21 and is highly alike to one coding protein of the receptor kinase. The TtLRR-STK gene can be inserted into the downstream of promoters with different types for building a plant expression vector and carrying out plant disease resistance improvement through the gene engineering technology, and moreover, the cloning of the gene can also be used for manufacturing the gene chip of the wheat and be applied to the researches of wheat disease resistance, and the like.

The invention discloses a mevalonate kinase gene RKMK and application thereof, the nucleotide sequence of the mevalonate kinase gene RKMK is shown as SEQ ID NO: 1, and the amino acid sequence coded by the gene is shown as SEQ ID NO: 2; the gene is separated from Rhodosporidium kratochvilovae YM25235, the gene is connected with a carrier and is transferred into a Rhodosporidium cell, and an experimental result shows that overexpression of the RKMK gene can promote synthesis of carotenoid by the Rhodosporidium kratochvilovae; according to the invention, the microorganisms are modified by means of genetic engineering, so that the yield of carotenoid in the microorganisms is increased, and a foundation is laid for large-scale commercialized production of carotenoid.

The invention belongs to the technical field of African swine fever treatment, and specifically relates to a new use of a compound ZL0580 for preventing or treating African swine fever. The inventionunexpectedly discovers that the compound ZL0580 can significantly inhibit the RNA and protein expression level of p30 and p72 in ASFV, prevents viruses from invading host cells, and can be used to inhibit the early infection of ASFV. The compound ZL0580 can significantly increase expression levels of genes such as TNF-alpha, NF-kappa B, IL-1beta, and IL-8 after ASFV infection, up-regulates transcription levels of TNF-alpha, NF-kappa B, IL-1beta, and IL-8, and enhances the immune response. Therefore, the compound ZL0580 can be used to prevent or treat African swine fever.

The invention discloses a method for establishing a canine distemper sensitive cell line based on a Nectin4 receptor. The method comprises the following steps: (1) extracting total RNA of a sample andsynthesizing cDNA; (2) amplifying and cloning Nectin4; (3) modifying Nectin4; (4) connecting and converting dNectin4-Hatag and extracting recombinant plasmid; (5) packaging recombinant slow virus; abd (6) transfecting cells and screening positive cell line. The Vero-Nectin4 established according to the method disclosed by the invention can stably express the Nectin4 receptor and is high in universality; the CDV virulent strain is capable of growing in the cell line and acquiring obvious CPE lesion; and the method is suitable for wide application in culturing or separating the canine distempervirus.

The invention provides an application of pyrimidine compounds in the promotion of metabolite synthesis and the hormone level in rice. The pyrimidine compounds are 6-(methoxymethyl)-2-[5- (trifluoromethyl)-2-pyridyl]pyrimidin-4-ol. The compounds can obviously rapidly improve the pathogen invasion resistance of plants, does not inhibit the growth of the plants or the growth of a root system, and also promotes the synthesis of the metabolites and the increase of the hormone content in the rice.