56 results about "Ankyrin" patented technology

Improved N-terminal capping modules for designed ankyrin repeat proteins (DARPins) conferring improved thermal stability to the DARPins are described, as well as nucleic acids encoding such proteins, pharmaceutical compositions comprising such proteins and the use of such proteins in the treatment of diseases.

New designed ankyrin repeat proteins with binding specificity for PDGF-BB are described, as well as nucleic acids encoding such PDGF binding proteins, pharmaceutical compositions comprising such proteins and the use of such proteins in the treatment of diseases.

The present invention provides stem cell feeder layer cell lines that contain are readily triggered to differentiation. The expression vector encodes the senescence-triggering factors (STFs) consisting of Cip / Kip, INK4A, Cy protein or ankyrin-binding protein motifs. Each expression vector also contains an inducible transcription regulation element for conditional expression of the STFs.

The present invention provides cell lines useful in the production proteins and peptides. The cell lines contain recombinant expression constructs. The recombinant expression construct encodes the STPs consisting of the Cy protein motif and / or an ankyrin-binding protein motif. Each recombinant expression construct also contains an inducible transcription regulation element having for conditional expression of the senescence-triggering factors (STPs).

The goal of the invention is to increase the therapeutical activity of oncolytic NDV. This issue is solved by a Newcastle Disease Virus comprising a recombinant nucleic acid, wherein the nucleic acid codes for a binding protein that has a therapeutic activity when expressed by the virus-infected tumor cell. Binding proteins belong to the following group: A natural ligand or a genetically modified ligand, a recombinant soluble domain of a natural receptor or a modified version of it, a peptide-ligand, an antibody molecule and derivatives thereof or antibody-like molecules like ankyrin repeat molecules or derivatives thereof.

The invention discloses recombinant escherichia coli for a cell surface display PET lytic enzyme, a construction and an application. A construction method thereof comprises the following steps: (1) chemically compounding ankyrin and PETase gene; (2) utilizing PCR to connect the ankyrin with the PETase gene, thereby acquiring a fused sequence; (3) connecting the fused sequence with an escherichia coli expression vector, thereby acquiring a fused expression vector; and (4) transferring the fused expression vector into expression host escherichia coli, thereby acquiring the recombinant escherichia coli for the cell surface display PET lytic enzyme. According to the construction method provided by the invention, the PETase with the objective protein having a PET degrading function is positioned to the cell surface through inaK-N, BrkA, AIDA or Lpp-OmpA ankyrin and has the characteristic of high catalytic activity, so that the problems of limited capacity of generating PETase, long growth period and low degradation rate of the wild bacterial strain can be solved; the enzyme does not need to be separated and purified; and the preparation and the use are simple.

Diacylglycerol (DAG) plays a central role in both the synthesis of complex lipids and in intracellular signaling; diacylglycerol kinase (DGK) catalyzes the phosphorylation of DAG, which yields phosphatidic acid. A family of DGKs has been identified in multicellular organisms over the past few years, but the physiological function(s) of this diversity is not clear. One clue has come from the Drosophila DGK2, rdgA, since mutations in this gene cause retinal degeneration. The present invention relates to a novel DGK, designated DGKι, which was isolated from human retina and brain libraries. DGKι contains two cysteine-rich repeats, a region similar to the phosphorylation site domain of MARCKS, a conserved catalytic domain, and four ankyrin repeats at its C-terminus. By primary structure, DGKι is most similar to human DGKζ and Drosophila rdgA. A>12 kb mRNA for DGKι was detected only in brain and retina among the tissues examined. In cells transfected with the DGKι cDNA, an approximately 130 kDa protein was detected by immunoassay, and activity assays demonstrated that it encodes a functional DAG kinase. The protein was found to be in both the cytoplasm and nucleus, with this localization controlled by PKC isoforms alpha and gamma. The gene encoding DGKι was localized to human chromosome 7q32.3-33, which is known to be a locus for an inherited form of retinitis pigmentosa. These results have defined a novel isoform of DAG kinase, which may have important cellular functions in the retina and brain.

The invention discloses a surface displaying system for rhizopus oryzaelipase, and a preparation method and application of the surface displaying system. According to the surface displaying system, a rhizopus oryzaelipase ProROL gene is connected to a C end of an ankyrin Pir1p gene; and by normal secretion and expression of an expression strain KM71H, a fusion protein is fixed on the surface of pichia pastoris KM71H strain by a covalent bond mode by using N-end repeated sequences of Pir1p as anchoring areas. By a gene fusion method, the rhizopus oryzaelipase gene and the C end of the mature ankyrin Pir1p are fused, the gene mpir1 is inserted into a vector containing different promoters pAOX1 and pGAP, recombinant plasmids AOX-mPir1 and GAP-mPir1 are obtained, the surface displaying of the pichia pastoris KM71H can be realized, and the high-efficiency expression can also be realized. Compared with the promoter pAOX1, the promoter pGAP has the advantages that the carbon source conversion is not required; the promoter pGAP is easy and convenient to prepare; methanol is not needed to be added in the induction process; the toxic and side effects of methanol on thallus growth are reduced; and the highest enzyme activity is 614.5U / L (180.74[dry cell weight]) and is greater than that of the yeast strain using the promoter pAOX1. The surface displaying system is high in practical applicability and has high economic and social benefit.

The invention discloses Pichia pastoris wall protein, a surface display system constructed thereby and a construction method thereof. The amino acid sequence of the Pichia pastoris wall protein is SEQ NO:1. The Pichia pastoris wall protein cell surface display system is formed by using the Pichia pastoris wall protein Gcw28 as ankyrin and fixing target protein on the surface of the Pichia pastoris cell. The expression quantity of the wall protein Gcw28 in the Pichia pastoris is very high and is respectively 8 times and 18 times higher than that of the endogenesis ankyrin Pir1 protein and Pir2protein applied to the Pichia pastoris surface display system.

New designed ankyrin repeat proteins with binding specificity for PDGF-BB are described, as well as nucleic acids encoding such PDGF binding proteins, pharmaceutical compositions comprising such proteins and the use of such proteins in the treatment of diseases.

The invention discloses novel application of an ankyrin repeat domain-containing 13A gene and / or a protein encoded by the same. An acute myeloid leukemia (AML) diagnosis and prognosis related molecular marker is screened to obtain an ankyrin repeat domain-containing 13A gene(ANKRD13A) closely related to AML patient prognosis; the high expression of ANKRD13A in the body of an AML patient warns poorprognosis; the ANKRD13A can be used as an independent factor for influencing the prognosis of the AML patient; the ANKRD13A is related to the FAB typing and the cytogenetics risk stratification of the AML patient; and the ANKRD13A may participate in the disease progression of AML through a variety of pathways. The ANKRD13A can be used for preparing a therapeutic drug or a prognosis detection reagent for the acute myeloid leukemia.

The invention relates to novel promoter sequences derived from a portion upstream of the coding sequence of the gene for the CARP protein (Cardiac Ankyrin Repeat Protein), and which are capable of controlling the level and the specificity of expression of a transgene in vivo in cardiac muscle cells. The invention thus describes novel compositions, constructs, vectors and their uses in vivo for the transfer and expression of a nucleic acid in vivo in cardiac muscle cells. The subject of the present invention is also the use of the promoter sequences for generating transgenic animals which constitute models for studying certain cardiac pathologies.

Provided herein are methods for stable integration and / or expression of one or more recombinant polynucleotides in a host cell. The recombinant polynucleotides are typically integrated into the host genome at some native chromosomal integration sites. The integration can be mediated by homologous recombination or by using a hybrid recombinase targeting the specific chromosomal locations. The native chromosomal integration sites in the host cells, which support stable integration and strong transcription activities of foreign genes, are present within or adjacent to specific genes in the CHO genome, ankyrin 2 gene (Ank2), cleavage and polyadenylation specific factor 4 gene (Cpsf4), C-Mos gene, and Nephrocystin-1 / Mal gene. Also provided are methods and nucleic acid molecules for inserting site-specific recombination sequences (chromosomal landing pads) into these specific chromosomal locations, engineered host cells containing chromosomal landing pads, methods and compositions (e.g., kits) therefore.

The invention discloses a double-lipase cell surface co-display engineering bacterium, and a construction method and application thereof. The engineering bacterium is co-displayed on the Pichia pastoris cell surface by using Saccharomyces cerevisiae cell wall protein Sed1p as ankyrin and Rhizopus oryzae lipase and Candida antarctica lipase B as double-lipase target proteins, thereby constructing the double-lipase cell surface co-display recombinant yeast engineering bacterium. The Rhizopus oryzae lipase and Candida antarctica lipase B are co-displayed on the Pichia pastoris cell wall surface to prepare the Rhizopus oryzae lipase / Candida antarctica lipase B cell surface co-display complete cell catalyst. The maximum enzyme activity of the surface co-display complete cell catalyst is 686U / g-cell dry weight, which is respectively 3 times or 2 times of the enzyme activity of the independently displayed Rhizopus oryzae lipase or Candida antarctica lipase B; and the methyl ester yield of the catalytic grease esterification reaction is up to 96%.

The present invention provides nucleic acids. Polypeptides, antibodies and cells related to MAIL (Molecule possessing Ankyrin repeats Induced by LPS; or IκB-ζ), a novel member of the IκB family, expressed in LPS stimulated monocytes and uses thereof. The isolated nucleic acids, polypeptides, antibodies and cells of the invention can be used in detection assays, screening methods and drug discovery assays.

New designed ankyrin repeat domains with binding specificity for serum albumin, recombinant binding proteins comprising at least two designed ankyrin repeat domains with binding specificity for serum albumin, as well as recombinant binding proteins comprising at least one designed ankyrin repeat domain with binding specificity for hepatocyte growth factor (HGF), at least one designed ankyrin repeat domain with binding specificity for vascular endothelial growth factor (VEGF-A), and at least two designed ankyrin repeat domain with binding specificity for serum albumin are described, as well as nucleic acids encoding such designed ankyrin repeat domains and recombinant binding proteins, pharmaceutical compositions comprising such designed ankyrin repeat domains, recombinant binding proteins or nucleic acids and the use of such designed ankyrin repeat domains, recombinant binding proteins, nucleic acids or pharmaceutical compositions in the treatment of diseases.

The present invention relates, generally, to inositol 1,4,5-triphosphate receptors (IP3R), and, in particular, to methods of modulating the activity of 220 kDa ankyrin-B in cellular localization and physiological function of IP3R. The invention further relates to methods of identifying compounds suitable for use in such methods and to compounds so identified.

The invention discloses Pichia pastoris wall protein, a surface display system constructed thereby and a construction method thereof. The amino acid sequence of the Pichia pastoris wall protein is SEQ NO:1. The Pichia pastoris wall protein cell surface display system is formed by using the Pichia pastoris wall protein Gcw51 as ankyrin and fixing target protein on the surface of the Pichia pastoris cell. The expression quantity of the wall protein Gcw51 in the Pichia pastoris is very high and is respectively 9 times and 21 times higher than those of the endogenesis ankyrin Pir1 protein and Pir2 protein applied to the Pichia pastoris surface display system.

The invention discloses a polymer microsphere and preparation thereof and application thereof in preparation of antibody purification filler. The polymer microsphere is prepared by the following steps of: taking polyvinyl alcohol and sodium alginate as raw materials, adding water to dissolve at 80-100 DEG C, cooling to 40 DEG C, and adding a recombinant escherichia coli solution containing ZZ fusion protein to prepare a water phase; and mixing liquid paraffin and a surfactant to form an oil phase, slowly dropwise adding the water phase into the oil phase, stirring at 30-70 DEG C to form emulsion, and adding a saturated boric acid calcium chloride aqueous solution to prepare the polymer microsphere, wherein the ZZ fusion protein is formed by fusing ZZ protein and ankyrin (LppOmpA) through a connecting peptide. The diameter of the polymer microsphere prepared by the method is 50-250nm, the ultra-large pore diameter is 1-2 microns, the raw materials are cheap, the cost is low, the operation is simple, the preparation processes of expression, purification, grafting and the like of conventional ligands are omitted, and the problems of expensive ligands and the like are solved.

The present invention relates to recombinant binding proteins comprising one or more designed ankyrin repeat domains with binding specificity for coronavirus spike proteins, nucleic acids encoding such proteins, pharmaceutical compositions comprising such proteins or nucleic acids, and the use of such proteins, nucleic acids or pharmaceutical compositions in the treatment of coronavirus diseases, particularly diseases caused by SARS-CoV-2.

The invention which belongs to the fields of gene engineering and gene diagnosis provides a method for detecting SNP of a BANK1 gene. The method comprises the following steps: 1, designing a primer and a probe for amplifying an rs10516487 site or an rs17266594 site; 2, carrying out PCR amplification by utilizing the primer and the probe with sample DNA as a template, wherein concentrations of the upstream primer and the downstream primer in the primer are different, and the concentration of one is not less than five times the concentration of other one; and 3, adding a saturated dye, analyzing a melting degree curve, and determining the SNP. The method which can detect the polymorphism of the rs10516487 site and the rs17266594 site of the BANK1 gene in peripheral blood or a body fluid can be used to determine the attack possibility of autoimmtme diseases of systemic lupus erythematosus, rheumatoid arthritis, systemic chorionitis and the like. The method has the advantages of rapidness, simplicity, no pollution, and high detection result resolution. The invention also provides a corresponding detection kit.

The present disclosure relates to multispecific proteins comprising designed ankyrin repeat domains with binding specificity for different targets, such as, e.g. CD40 and FAP. In addition, the present disclosure relates to nucleic acids encoding such multispecific proteins, pharmaceutical compositions comprising such multispecific proteins or nucleic acids, and the use of such binding proteins, nucleic acids or pharmaceutical compositions in methods for treating or diagnosing diseases, such as cancer, in a mammal, including a human.