50 results about "Cellular localization" patented technology

The present invention is directed to a method for detection, characterization and isolation of nucleic acids encoding proteins of a desired property, such as a particular cellular localization. The invention further provides for rapid expression of such proteins or glycoproteins in mammalian cells and for facilitated purification of the novel secreted proteins or glycoproteins. Further, the invention provides for radioactive labelling of the novel proteins or glycoproteins, for rapid identification of sites of binding including animals and for rapid production of infective viral vectors for use in gene transfer.

The invention relates to cellular localization signals. In particular, the invention relates to endoplasmic reticulum localization signals in monomeric or multimeric form. The localization signals are utilized as research tools or are linked to therapeutics. Disclosed are methods of making and using polypeptides and modified polypeptides as signals to localize therapeutics, experimental compounds, peptides, proteins and / or other macromolecules to the endoplasmic reticulum of eukaryotic cells. The polypeptides of the invention optionally include linkage to reporters, epitopes and / or other experimental or therapeutic molecules. The invention also encompasses polynucleotides encoding the localization signals and vectors comprising these polynucleotides.

The invention relates to kinase inhibitor ligands and polyligands. In particular, the invention relates to ligands and polyligands that modulate ERK activity. The ligands and polyligands are utilized as research tools or as therapeutics. The invention includes linkage of the ligands and polyligands to a cellular localization signal, epitope tag and / or a reporter. The invention also includes polynucleotides encoding the ligands and polyligands.

The invention provides a cellular localization unit, array and device and a formation method of the cellular localization unit, array and device. The cellular localization unit comprises a substrate and a cellular localization hole. At least one pair of microelectrodes is arranged on the substrate; the microelectrodes are separated and each pair of the electrodes is symmetrically dispersed relatively to the same point to generate a normal dielectrophoretic force area at an area where the ends of the microelectrodes gather; and voltage signals connected to adjacent microelectrodes have opposite phases. The cellular localization hole is located above the microelectrodes; the accommodation room of the cellular localization hole exposes the normal dielectrophoretic force area; and the accommodation room is for a single cell. AC signals with certain amplitudes, frequencies and phases are applied to the microelectrodes, so that cells move towards the surfaces of the microelectrodes, and the cellular localization hole matches the microelectrodes to allow a single cell to be accurately localized at a certain position.

Previously we have identified a conserved domain (SAP, for SAF-A / B, Acinus, and PIAS) in the foot-and-mouth disease virus (FMDV) leader (L) protein coding region that is required for proper sub-cellular localization and function. Mutation of isoleucine 55 and leucine 58 to alanine (I55A, L58A) within the SAP domain resulted in a viable virus that displayed a mild attenuated phenotype in cell culture, along with altered sub-cellular distribution of L and failure to induce degradation of the transcription factor nuclear factor kappa-B. Here we report that inoculation of swine and cattle with this mutant virus results in the absence of clinical disease, the induction of a significant FMDV-specific neutralizing antibody response, and protection against subsequent homologous virus challenge. Remarkably, swine vaccinated with SAP mutant virus are protected against wild type virus challenge as early as two days post-vaccination suggesting that a strong innate as well as adaptive immunity is elicited. This variant could serve as the basis for construction of a live-attenuated FMD vaccine candidate.

Reagents, methods, and kits for the classification of cancer that comprise or employ antibodies that bind specific regions of CD43. One method includes contacting tissue with an antibody capable of specifically binding the cytoplasmic tail of CD43, contacting the tissue with an antibody capable of specifically binding the extracellular domain of CD43, and resolving cellular localization of any binding to the tissue with the antibody capable of specifically binding the cytoplasmic tail of CD43 and the antibody capable of specifically binding the extracellular domain of CD43. The binding patterns of the antibodies can be used to characterize cancer as more aggressive or less aggressive and can distinguish small cell lung cancer from non-small cell lung cancer. The cancer may therefore be treated in accordance of the characterization.

The present invention provides a novel PrP protein, and nucleic acids encoding this protein, where the PrP protein is characterized in vivo by 1) incomplete glycosylation relative to glycosylation of wild-type PrP<C >and 2) proper cellular localization, i.e. an ability to be transported to the cell surface. This novel, under-glycosylated PrP, unlike its normal cellular counterpart, can easily be converted into a protease-resistant isoform by incubation with infectious prions. The invention further provides systems for the study of prion disorders and methods of using these systems, e.g. the study of the mechanical processes in progression of prion-mediated disease or the identification of new therapeutic agents for treatment of prion-mediated disorders. In such systems, protease-resistant under-glycosylated PrP is generated de novo and can be detected by standard immunoblot techniques.

The invention discloses a flow cytometry electrofusion apparatus. The flow cytometry electrofusion apparatus comprises: a one-to-one cell fusion portion, a fusion cell detecting and sorting portion and a single cell collection portion. The one-to-one cell fusion portion is provided with a plurality of parallel cellular localization channels. Different cells can be adsorbed to ends of the channels, and two different cells can close to each other and be fused. The fused cells are placed until membranes of the different cells are completely fused, and then are put into the fusion cell detecting and sorting portion. The fusion cell detecting and sorting portion comprises a laser light source, a fluorescence detector and a deflector. The fused cells, i.e., red and green double fluorescent stained cells, are collected in a collecting tube. When collecting a fused cell, flow of droplet in the fluorescence detector is stopped. 200 [mu]l of cell culture medium is added to the collecting tube through a cell culture medium accumulator installed above the collecting tube to open the end of the collecting tube. The 200 [mu]l of the cell culture medium containing the fused cell flows into a well of a 96-well plate arranged blow the collecting tube. At this time, the flow of the liquid in a flow chamber is started, and the next fused cell is put into the next well of the 96-well plate. According to the present invention, one-time continuous accomplishment of one-to-one cell fusion can be realized, fusants can be detected correctly after fusing and each fusant can be cultured respectively.

With the goal of creating a combination vaccine against Shigella and other diarrheal pathogens we have constructed a prototype vaccine strain of Shigella flexneri 2a (SC608) that can serve as a vector for the expression and delivery of heterologous antigens to the mucosal immune system. SC608 is an asd derivative of SC602, a well-characterized vaccine strain, which has recently undergone several phase 1 and 2 trials for safety and immunogenicity. Using non-antibiotic asd-based plasmids, we have created novel constructs for the expression of antigens from enterotoxigenic E. coli (ETEC), including CFA / I (CfaB and CfaE) and the B-subunit from heat-labile enterotoxin (LTB) in Shigella vaccine strain SC608. Heterologous protein expression levels and cellular localization are critical to immune recognition and have been verified by immunoblot analysis. Following intranasal immunization (SC608(CFAI) and SC608(CFAI / LTB) of guinea pigs, serum IgG and IgA immune responses to both the Shigella LPS and ETEC antigens can be detected by ELISA. In addition, ELISPOT analysis for ASCs from cervical lymph nodes and spleen showed similar responses. All vaccine strains conferred high levels of protection against challenge with wild-type S. flexneri 2a using the Sereny test. Furthermore, serum from guinea pigs immunized with SC608 expressing CfaB and LTB contained antibodies capable of neutralizing the cytological affects of heat-labile toxin (HLT) on Chinese Hamster Ovary (CHO) cells. These initial experiments demonstrate the validity of a multivalent invasive Shigella strain that can serve as a vector for the delivery of pathogen-derived antigens.

The invention belongs to the technical field of plant tissue and cell culture, and particularly relates to a preparing method of carnation leaf protoplast. With a carnation tissue culture seedling asthe test material, an efficient protoplast preparing method for purification through a low-speed oscillation enzymolysis-sedimentation method is established. The method is convenient to operate and can be implemented all year round, the yield of the protoplast subjected to low-speed oscillation enzymolysis separation and density gradient purification is high and can reach 183*105 p / gFW, the cell integrity is high, and it is ensured that the activity is 75% or above. By means of the method, a foundation is laid for the research on the aspects of gene function authentication, sub-cellular localization, protein interaction and the like of a carnation leaf instantaneous converting system.

The invention relates to the field of longevity enhancement. More particularly, the invention provides compositions and methods relating to CRTC modulation. In certain embodiments, the invention provides compositions and methods for enhancing longevity in an organism by inhibiting CRTC activity, such as, for example, inhibiting CRTC expression or cellular localization in the organism.

Methods for detecting, diagnosing and monitoring an epithelial cancer in a patient are described comprising measuring in a sample from the patient Ep-ICD polypeptides and Ep-ICD polynucleotides. Methods for prognosis of breast cancer comprising measurement of nuclear Ep-ICD polypeptides and optionally EpEx polypeptides are provided. The invention also provides kits and compositions for carrying out the methods of the invention. The invention also provides a unique scoring system using immunohistochemical analysis to arrive at an Ep-ICD Subcellular Localization Index (ESLI) score, which is used to arrive at a diagnosis of cancer in a patient, or, more particularly, to identify patients that are in need of aggressive clinical treatment.

The invention discloses a flow cytometry electrofusion apparatus. The flow cytometry electrofusion apparatus comprises: a one-to-one cell fusion portion, a fusion cell detecting and sorting portion and a single cell collection portion. The one-to-one cell fusion portion is provided with a plurality of parallel cellular localization channels. Different cells can be adsorbed to ends of the channels, and two different cells can close to each other and be fused. The fused cells are placed until membranes of the different cells are completely fused, and then are put into the fusion cell detecting and sorting portion. The fusion cell detecting and sorting portion comprises a laser light source, a fluorescence detector and a deflector. The fused cells, i.e., red and green double fluorescent stained cells, are collected in a collecting tube. When collecting a fused cell, flow of droplet in the fluorescence detector is stopped. 200 [mu]l of cell culture medium is added to the collecting tube through a cell culture medium accumulator installed above the collecting tube to open the end of the collecting tube. The 200 [mu]l of the cell culture medium containing the fused cell flows into a well of a 96-well plate arranged blow the collecting tube. At this time, the flow of the liquid in a flow chamber is started, and the next fused cell is put into the next well of the 96-well plate. According to the present invention, one-time continuous accomplishment of one-to-one cell fusion can be realized, fusants can be detected correctly after fusing and each fusant can be cultured respectively.

A method of determining whether a subject is at risk of developing osteoarthritis (OA), said method comprising: determining the cellular localization of a Prohibitin-1 (PHB1) polypeptide and / or Small Ubiquitin-like Modifier (SUMO) polypeptide and / or UBC9, in a cell sample from said subject; and determining whether said subject is at risk of developing OA based on the cellular localization of a PHB1 polypeptide and / or SUMO and / or UBC9 polypeptide, is described.

Methods and kits for the prognosis of breast cancer comprising measurement of nuclear Ep-ICD poly-peptides are provided. Measurement may be quantitative and / or qualitative. The invention also provides a system for generating an Ep-ICD Subcellular Localization Index (ESLI) value, which may be used to prognose breast cancer in a subject.

The invention provides a pawpaw functional centromere antigen polypeptide, and applications thereof. The amino acid sequence of pawpaw functional centromere histone CENH3 is represented by SEQ ID No.2; a specific sequence of the N terminal of the amino acid sequence coded based on the pawpaw functional centromere histone CENH3 is used for synthesis of a polypeptide; routine immune-inoculation of New Zealand white rabbits with the polypeptide is carried out, antiserum is prepared, the antibody of CENH3 protein is obtained via antigen affinity purification, and immunofluorescence assay of the antibody is carried out. The antibody of CENH3 protein can be used for cellular localization of pawpaw functional centromere, and can be applied to pawpaw chromatin immunoprecipitation assay sequencing (ChIP-seq), so that the position, size, and sequence information of functional centromeres on pawpaw chromosomes can be determined, obtained physical map information is more complete, and centromere-related information can be provided for pawpaw genome assembly.