137results about How to "Improve expression level" patented technology

The present invention provides novel methods and compositions for the diagnosis and treatment of solid cancers. The invention also provide methods of identifying inhibitors of tumorigenesis.

The present invention provides a cell culture medium formulation that supports the in vitro cultivation, particularly in suspension, of mammalian cells, particularly epithelial cells and fibroblast cells, and methods for cultivating mammalian cells in suspension in vitro using these media. The media comprise a basal medium and a polyanionic or polyanionic compound, preferably a polysulfonated or polysulfated compound, and more preferably dextran sulfate. The present invention also provides chemically defined, protein-free eukaryotic cell culture media comprising an iron chelate and zinc, which is capable of supporting the growth (and particularly the high-density growth of mammalian cells) in suspension culture, increasing the level of expression of recombinant protein in cultured cells, and/or increasing virus production in cultured cells.

Methods of shuffling DNA to obtain recombinant herbicide tolerance nucleic acids encoding proteins having new or improved herbicide tolerance activities, libraries of shuffled herbicide tolerance nucleic acids, transgenic plants and DNA shuffling mixtures are provided.

The present invention provides an agent which is effective to a patient who is administered by an agent comprising anti-CD20 antibody as an active ingredient.

A method of amplifying a template DNA molecule comprising incubating the template DNA molecule in a reaction mixture comprising a DNA polymerase and at least one accessory protein at a constant temperature to produce amplified product, wherein production of amplified product does not require exogenously-added oligonucleotide primers and the template DNA molecule does not have have terminal protein covalently bound to either 5′ end.

The present invention provides methods comprising both genetic and chemical means for the production of hepatocytes from a variety of cell sources, particularly pluripotent stem cells.

Provided herein are compositions and methods for cancer diagnosis, research and therapy, including but not limited to, cancer markers. In particular, provided herein are non-coding RNAs as diagnostic markers and clinical targets for cancer.

An isolated polynucleotide functional as a promoter in eukaryotic cells is disclosed. The isolated polynucleotide includes an endothelial specific enhancer element as detailed herein. Further disclosed is a method of expressing a nucleic acid sequence of interest in endothelial cells.

A population of enteroendocrine cells (EEC) is obtained from a mammalian post-natal cell population, such as a population including post-natal stem cells, by treating the population with a plurality of small molecules that upregulate ChgA and promote differentiation of the cells to form the enteroendocrine cells. The upregulation of ChgA is such that the fraction of cells expressing CGA in the obtained cell population, as measured by a ChgA Immunostaining Assay, is at least about 1.5%. Small molecules that can be used to differentiate the post-natal cells into the enteroendocrine cells can include at least one of a Wnt activator, a Notch inhibitor, a Wnt inhibitor, a MEK/ERK inhibitor, a growth factor, a HDAC inhibitor, a Histone Methylation Inhibitor, a Tgf-β inhibitor, and a NeuroD1 activator. Also, the insulin expression of a population of mammalian cells is increased by treating the population with a plurality of small molecules that increase the insulin expression.

The present invention is directed to isolated nucleic acid molecules encoding a voltage gated sodium channel of a peripheral nerve cell, as well as to the isolated voltage gated sodium channels of a peripheral nerve cell encoded thereby. Methods for increasing or decreasing the expression of functional voltage gated sodium channels in host cells are also provided, as well as methods using the sodium channels. Also provided is a method for isolating other voltage gated sodium channels.

This invention discloses fusion protein of soluble receptor II of human tumor necrosis factor and Fc fragment of antibody. The amino acid sequence of the fusion protein is shown in SEQ ID No.2. This invention utilizes IRES sequence of double-cistronic vector pTandem-1 to ligate the gene sequence of the fusion protein with DHFR gene, which is used as a screening marker. This invention can reduce the screening intensity and raise the expression level. This invention can effectively and stably produce fusion protein of soluble receptor II of human tumor necrosis factor and Fc fragment of antibody.

The invention relates to a respiratory syncytial virus vaccine, and the preparation method and application, in particular to an application of escherichia coli to express and recombinant respiratory syncytial virus G protein and mutation G protein, and preparation vaccine with nontoxic typed escherichia coli heat labile enterotoxin, for human or animal preventive inoculation, and for respiratory syncytial virus resistance, belonging to the field of biotechnology. The respiratory syncytial virus vaccine is characterized in that: the respiratory syncytial virus subunits vaccine comprises lopped respiratory syncytial virus RSV protein G, and further comprises nontoxic typed escherichia coli heat labile enterotoxin LT adjuvant. The vaccines are lopped respiratory syncytial virus RSV protein G containing amino acid between aa130 and 230 of the original G protein, and substitutes the amino acid CAWIC (CX3C module ordered) between aa182 and 186 with the amino acid YLEKESIYY (CTL epitope) on the RSV M protein, forming the GCIL protein. The respiratory syncytial virus vaccine has the advantages of remarkable practical significance for preventing human respiratory syncytial virus infection.

A cell of the present invention contains a nucleic acid construct encoding a WT1 gene product or a fragment of the WT1 gene product. The nucleic acid construct contains (i) a region encoding a desired fragment of the WT1 gene product and (ii) only AUG as a functional start codon. The present invention can provide a cell into which the nucleic acid construct is introduced so that an expression level of a WT1 gene product or a fragment of the WT1 gene product is remarkably enhanced.

The invention provides a mutant cephalosporin C acylase. One or a plurality of amino acid locus replacement is carried out on an amino acid sequence shown as SEQ ID NO.1 in a sequence table to obtain a mutant amino acid sequence; and one or more of 288-locus valine, 296-locus histidine and 417-locus histidine is or are adopted as amino acid locus(es). The technical problems that the mutant cephalosporin C acylase of the prior art has small catalytic activity, seriously influenced enzymatic activity by the outside and product inhibition are solved.

The invention provides a method for realizing gene transient expression in peach fruits. According to the method, genes are excessively expressed in the pulp tissue through a vacuum infiltration infection method, and the method can be used for researching the gene functions. The method comprises the following steps: (1) selecting the ripeness of the peach fruits; (2) carrying out competence culture on the peach pulp tissue for pre-infection; (3) soaking the pulp in an agrobacterium infection solution containing a gene sequence; (4) applying vacuum to the pulp tissue; (5) releasing the vacuum for realizing transgenosis; and (6) carrying out further culture on the pulp tissue after infection. The established gene transient expression method is suitable for the peach fruits with thin peel and without cavities in the pulp tissue, the operation is simple, the expression level is high, the result is reliable, and a technical support is provided for developing gene function identification and selecting functional genes.

Methods for identifying a cancer patient, such as a breast cancer patient, suitable for treatment with a 4-anilinoquinazoline kinase inhibitor, such as lapatinib, and an AKT inhibitor, comprising detecting modulated expression of HER2 (ERBB2) and SASH1 or protein encoded thereof and detecting PIK3CA mutation status. High levels of expression in HER2 and high levels of SASH1 and/or positive PIK3CA mutation status indicate a patient that is suitable for treatment with a 4-anilinoquinazoline kinase inhibitor, such as lapatinib and an AKT inhibitor.

The embodiments of the present invention are directed to discrete, cis-acting regulatory elements that include a barrier element, an insulating element, a silencing element, and matrix attachment regions (“MARs”). Additional embodiments of the present invention are directed to nucleic acid molecules that are useful for facilitating stable transgene expression within a chromatin environment. Additional embodiments of the present invention are directed to recombinant expression vectors including nucleic acid molecules of the present invention that can be incorporated into artificial chromosomes, eukaryotic cell-lines, non-human transgenic animals, and transgenic plants, to improve recombinant protein production in a broad range of eukaryotic hosts, and pharmaceutical compositions including nucleic acid molecules of the present invention that are also useful for gene therapy in the treatment of various genetic diseases.

The invention discloses a gene synthesis of Alpha-interferon of wild boars and vector construction thereof as well as a production method of a product. A number of problems of low expression capacity, products expressed in a fusion state, products with purification tags or no fermentation technology with high density exist in the domestic recombinant strains. The invention includes a method that a codon and codon pairs, preferred by Escherichia coli, are used for synthesizing an Alpha-interferon gene of the wild boar, establishing a high-efficiency expression vector and transforming a high-efficiency expression strain as well as methods of high-density fermentation of engineering bacteria, separation and purification of inclusion bodies, the modification, the renaturation and the purification of target protein and the determination of biological activity of the expressed product. The gene synthesis, the vector construction and the production method pertain to the technical field of the production of polypeptide drugs by genetic engineering in biopharmaceuticals.

The invention discloses a silkworm PABP (polyadenylic acid bonding protein) bonding protein interacting factor gene BmPaip1. The nucleotide sequence of the gene BmPaip1 is shown as SEQIDNO.3, the open reading frame of the gene BmPaip1 is 1194bp long, composed of eight exons and codes 398 amino acids, theoretical protein molecular weight is 44kDa, and isoelectric point pI is equal to 5.02; and SMART online software analysis shows that the bits from 100 to 398 in the amino acid sequence of the gene BmPaip1 of a silkworm are highly homologous with an eIF4G protein of an eucaryon, and expression detection in insect cells shows that expression level of a reporter gene LUC (luciferase) can be obviously improved in cells genetically modified by the gene BmPaip1 of the silkworm, so that the gene BmPaip1 of the silkworm can be combined with eIF4E and eIF4A to form a translation initiation complex and can be used for improving translation efficiency of an exogenous gene.

The invention discloses application of long-chain non-coding RNA NONHSAG039951.2 as a diagnostic marker for osteoporosis. Through detection, it is found that the LncRNA NONHSAG039951.2 is expressed inbone tissue of both osteoporosis patients and normal persons, the expression level of the LncRNA NONHSAG039951.2 in the bone tissue of the osteoporosis patients is significantly higher than that of the normal persons, the difference has statistical significance, and it is suggested that the LncRNA NONHSAG039951.2 as the diagnostic marker for the osteoporosis has feasibility and clinical application value. The long-chain non-coding RNA NONHSAG039951.2 can be used as the diagnostic marker for the osteoporosis and achieves the purposes of early diagnosis and early treatment of the osteoporosis.

The invention applies to the technical field of biological medicine and provides a method for infecting infecting mesenchymal stem cells by Runx2 recombinant lentiviruses to express osteoblastic genes. The method comprises steps as follows: Runx2 gene extraction and plasmid construction are performed; the Runx2 recombinant lentiviruses are packaged; the Runx2 recombinant lentiviruses infect the mesenchymal stem cells, so that infected mesenchymal stem cells capable of expressing the osteoblastic genes are obtained.

The present invention discloses recombinant escherichia coli and an application thereof. The recombinant escherichia coli is deposited on August 28, 2018 in the China General Microbiological Culture Collection Center and has deposit number of CGMCC No.16349. After the recombinant escherichia coli is fermented and cultured, concentration of Microcin J25 in supernatant can reach 4,000 mg/L or more.

A method of treatment is disclosed. The method comprises administering to a human subject, known to have decreased glutathione activity, an effective amount of a nitroxide antioxidant, wherein the nitroxide antioxidant increases an expression level of one or more genes encoding glutathione S-transferase enzymes, thereby increasing glutathione activity.

Disclosed herein is a novel apparatus and the uses thereof in the prophylaxis and/or treatment of neuropsychiatric disorders. The present apparatus comprises a detecting means, a stimulation means, a virtual reality means and a processor. According to some embodiments of the present disclosure, the present apparatus produces an additive or synergistic effect on the treatment of neuropsychiatric disorders.

The invention relates to a CYP101 enzyme recombinant vector, a construction method thereof and a CYP101 enzyme high-efficiency expression and purification method, and belongs to the field of biotechnology. A PET28a-CYP101 high-efficiency expression vector is constructed and transformed into BL21 plysS strain, and high-purity enzyme with purity higher than 95% is obtained by a simple two-step purification method including affinity chromatography and ion exchange chromatography after inducible expression. Compared with the prior art, CYP101 is simply and efficiently produced, the cost is low, purification is facilitated, and 23mg of target protein with the purity higher than 95% can be obtained by per liter of culture media. The CYP101 enzyme high-efficiency expression and purification method can be used for expressing and purifying CYP101 enzyme, the production efficiency of a laboratory is greatly improved, and experimental cost is greatly reduced. The CYP101 enzyme high-efficiency expression and purification method has an excellent application prospect for large-scale CYP101 enzyme production and purification.

Contemplated compositions and methods are drawn to biomarkers and methods related to treatment of neoplastic disease with Hec1 inhibitor. Gene status and/or expression levels of Hec1(HEC), Rb(RB1), and/or p53 (TP53) may be useful as biomarkers for sensitivity to treatment with a Hec1 inhibitor. In addition, Hec 1 inhibitors may show synergistic effects when used in conjunction with cytotoxic drugs.

The present invention provides proteins having the Na⁺/glucose transporter activity, DNAs encoding the proteins, a method of screening for a compound enhancing or inhibiting the activity of the proteins, and compounds obtained by the screening method. The proteins having the same or substantially the same amino acid sequence as the amino acid sequence represented by SEQ ID NO: 1, 15 or 26 are useful as a diagnostic marker for a disease such as diabetes. The compounds enhancing or inhibiting the activity of the proteins, obtained the screening method using the proteins, can be used as a prophylactic and/or therapeutic agent for a disease such as diabetes and hyperlipidemia.

The invention relates to compositions, kits, and methods for detecting, characterizing, preventing, and treating human breast cancers. A variety of newly identified markers are provided, wherein changes in the levels of expression of one or more of the markers is correlated with the presence of breast cancer.

The invention relates to a recombinant DNA vector for efficiently preparing foot and mouth disease virus-like particles, the foot and mouth disease virus-like particles prepared by utilizing a recombinant expression plasmid vector containing the recombinant DNA vector to transfect a host cell, application thereof and a vaccine. Aiming at the problems of toxicity of 3C protease to the host cell andimpact of 3C protease on yield of foot and mouth disease VLPs, a 3CshRNA segment having disturbing effect on gene expression of 3C protease is designed, so that expression of 3C protease in the process of preparing the foot and mouth disease VLPs is reduced obviously; three sites are mutagenized on the basis of original foot and mouth disease virus 3C protease gene, so that action effect of 3C protease is regulated specifically. Through modifying the 3C protease gene, the toxicity of 3C protease to the host cell is lowered, and effective cutting performance of 3C protease to foot and mouth disease virus P1 precursor protein is improved, so that synthesis yield of the foot and mouth disease VLPs is increased greatly, and a foundation is laid for large-scale production and use of foot and mouth disease VLP vaccines.

The invention relates to an artificially synthesized signal peptide and application thereof. An amino acid sequence of the signal peptide is selected from any one of SEQIDNO: 1-5. The signal peptide can be added in a mammal zooblast expression vector for guiding a mammal source protein to secrete and express, and can improve the exogenous protein expression in a zooblast by 2-36 times.