Abstract: Disclosed are mutant DNA polymerases having improved extension rates relative to a corresponding, unmodified polymerase. The mutant polymerases are useful in a variety of disclosed primer extension methods. The mutant polymerases overcome the inhibitory effects of a variety of polymerase and reverse transcriptase inhibitors. Therefore, the mutant polymerases are useful in a variety of disclosed methods in the presence of such inhibitors.

Abstract: The present invention relates to a method for purifying a virus, or a viral antigen thereof, comprising at least the following steps: a) obtaining a fluid comprising the virus, or a viral antigen thereof, and b) purifying the fluid by at least one density gradient ultracentrifugation step, wherein the ratio of the amount of virus, or viral antigen thereof, present in the fluid over the density gradient volume is less than 1, less than 0.8, less than 0.6 and less than 0.4.

Abstract: Disclosed is a system for detecting a protease inside a cell. In one embodiment, the system includes a chimeric protein that comprises as covalently linked components: 1) at least one optionally masked signal protein; 2) at least one protease-specific cleavage site; and 3) at least one detectable amino acid sequence. The invention has a wide spectrum of applications including use in the detection of novel protease inhibitors inside cells and tissue.

Abstract: The present invention provides improved systems and methods for amplifying nucleic acids. Among other things the present invention provides a system for amplifying nucleic acids through use of a primase and a polymerase with strand-displacement ability without, for example, exogenously-added primers. The present invention is particularly useful for whole genome amplification.

Abstract: A reversibly modified ‘hot start’ RNAse H enzyme composition is described for the improved CATACLEAVE™ probe detection of nucleic acid sequences in a test sample. A key feature of the enzyme composition is the ability to regulate the catalytic activity of the RNAse H during the course of a reverse transcription-PCR cycle. Thus, RNAse H activity can be initially suppressed to minimize degradation of RNA:DNA primer heteroduplexes prior to reverse transcription. After cDNA synthesis is complete, RNAse H activity is induced to promote the cleavage and fluorescent detection of CATACLEAVE™ probes that anneal to target DNA sequences within the reverse transcriptase-PCR products. The inducible RNAse H enzyme is amenable to high throughput applications requiring one step reverse transcriptase CATACLEAVE™ PCR in a single reaction mix.

Abstract: A method of amplification and/or of detection for removing contaminants in a liquid biological sample containing nucleic acids of interest to amplify, which includes treating the biological sample chemically or enzymatically to convert one type of base of the nucleic acids of interest to another type of base; adding amplification primers, intended for specifically amplifying said converted nucleic acids of interest, each primer being constituted of three different types of bases; adding to the biological sample, after these treatments, amplification reagents the primers previously synthesized; and placing the solution and the reagents in conditions permitting amplification and/or detection of the converted nucleic acids.

Abstract: Compositions, methods, and kits for detecting and monitoring kinase, phosphatase and protein post-translational modification activity are described. The compositions typically include a peptide, a detectable moiety, and a protease cleavage site. Modification of a peptide by a kinase, phosphatase or other protein post-translational modification alters the proteolytic sensitivity of the peptide, resulting in a change of a detectable property of the composition. Panel assays for determining substrates or modulators of kinase, phosphatase or other protein post-translational modification activity are also described.

Abstract: This invention provides a class of compounds which are useful for specifically inhibiting cyclin-dependent kinases. This class of compounds finds use in treating diseases resulting from inappropriate activity of cyclin-dependent kinases, including cancer, viral infections (e.g., HIV) neurodegenerative disorders (e.g. Alzheimer's disease), and cardiovascular disorders (e.g. atherosclerosis). Moreover, certain members of this class are particularly useful for inhibiting cyclin-dependent kinase 7 and are especially useful for the treatment of breast cancer.

Abstract: The invention provides a means for reconstituting I?B kinase in yeast in order to study the structure and regulation of IKK and to produce pharmacological therapies to block IKK. This invention can be used to express an IKK complex that is biochemically identical to IKK isolated from native cells and in coupled in vitro kinase assays to screen for its upstream regulators. The IKK expressed by reconstituting the yeast can be used to screen for unknown substrates and for pharmacological therapies that block its activity. The invention could also be used to screen for compounds that enhance its activity. The IKK may also be used as a source of material for crystallization and X-ray diffraction analysis.

Abstract: The invention provides the identification of the presence of mutant ROS protein in human cancer. In some embodiments, the mutant ROS are FIG-ROS fusion proteins comprising part of the FIG protein fused to the kinase domain of the ROS kinase. In some embodiments, the mutant ROS is the overexpression of wild-type ROS in cancerous tissues (or tissues suspected of being cancerous) where, in normal tissue of that same tissue type, ROS is not expressed or is expressed at lower levels. The mutant ROS proteins of the invention are anticipated to drive the proliferation and survival of a subgroup of human cancers, particularly in cancers of the liver (including bile duct), pancreas, kidney, and testes. The invention therefore provides, in part, isolated polynucleotides and vectors encoding the disclosed mutant ROS polypeptides (e.g., a FIG-ROS(S) fusion polypeptide), probes for detecting it, isolated mutant polypeptides, recombinant polypeptides, and reagents for detecting the fusion and truncated polypeptides.

Abstract: A process is disclosed for the production of cyclic di-guanosine monophosphate (c-di-GMP) without the use of protecting groups by means of an enzymatic synthesis. The process comprises the coupling of two guanosine triphosphate (GTP) molecules so as to form a c-di-GMP molecule. This is done under the influence of a mutant diguanylate cyclase (DGC) comprising the amino acid sequence V153M154G155G156. It has been found that the DGC is obtainable from inclusion bodies, and therewith can be made available in amounts sufficient to improve the c-di-GMP synthesis. Particularly, the latter synthesis can be conducted in a one-pot method starting from commercially available bulk chemicals and allows upscaling to a commercial production scale.

Abstract: Methods and kits for efficient amplification of nucleic acids are provided. The methods comprise in-vitro amplification of a nucleic acid template employing partially constrained primers having terminal mismatch primer-dimer structure. The methods also comprise in-vitro amplification of a nucleic acid template employing partially constrained primers having nucleotide analogues. The methods enhance efficiency of nucleic acid amplification reaction by reducing non-specific amplification reactions.

Abstract: Gene encoding human glucokinase mutant is provided. The gene has the nucleotide sequence chosen from the nucleotide sequence listed as SEQ ID NO:2 and the nucleotide sequence wherein the ORF region encodes the same amino acid sequence as the one encoded by ORF region (position 487 to 1884) of SEQ ID NO:2 and the rest of the region is same as the non-ORF region of SEQ ID NO:2. Human glucokinase mutant encoded by the gene, the recombinant vectors carrying the gene, the hosts comprising the vectors, pharmaceutical compositions thereof, uses thereof, and methods for treating and preventing diseases by using the same are provided. The human glucokinase mutant encoded by the gene has higher activity than that of the wild type human glucokinase, and thus provides a new way of controlling blood glucose and/or preventing and/or treating disturbance of carbohydrate metabolism, especially preventing and treating diabetes.

Abstract: Methods of isolating target double-stranded polynucleotides with internal single-stranded regions are provided.

Abstract: The present invention is directed to methods of producing recombinant functional heme-binding proteins with complete heme incorporation and purified preparations of the same. The present invention is further directed to methods of identifying agents that modulate the activity of heme-binding proteins.

Abstract: The present invention provides fusion proteins including a green fluorescent protein inserted into the internal amino acid sequence of a G?s protein and further provides method of using the fusion protein construct to follow activation of a G-protein receptor by a candidate drug.

Abstract: A novel gene encoding P. pastoris orotate-phosphoribosyl transferase (URA5) is disclosed. Methods for producing and selecting yeast strains capable of stable genetic integration of heterologous sequences into the host genome are also provided.

Abstract: The present invention provides improved DNA polymerases, in particular, type A DNA polymerases, that may be better suited for applications in recombinant DNA technologies. Among other things, the present invention provides modified DNA polymerases derived from directed evolution experiments designed to select mutations that confer advantageous phenotypes under conditions used in industrial or research applications.

Abstract: The present invention provides thermostable enzymes, such as DNA polymerases and restriction endonucleases, that are substantially free from contamination with nucleic acids. The invention also provides methods for the production of these enzymes, and kits comprising these enzymes which may be used in amplifying or sequencing nucleic acid molecules, including through use of the polymerase chain reaction (PCR).

Abstract: The invention relates to an antibody specific to Erk1/2 phosphorylated at Thr188 and a method for producing the same. The invention also relates to an in vitro method for determining the presence of phosphorylated Erk1/2 in a sample using the antibody of the invention. Therefore, the invention also comprises an assay for diagnosing a heart disease in vitro comprising the antibody of the invention and certain uses of the antibody. Moreover the invention relates to a peptide used to produce the antibody.

Abstract: The present invention concerns methods and compositions for forming immunotoxin complexes having a high efficacy and low systemic toxicity. In preferred embodiments, the toxin moiety is a ranpirnase (Rap), such as Rap(Q). In more preferred embodiments, the immunotoxin is made using dock-and-lock (DNL) technology. The immunotoxin exhibits improved pharmacokinetics, with a longer serum half-life and significantly greater efficacy compared to toxin alone, antibody alone, unconjugated toxin plus antibody or even other types of toxin-antibody constructs. In a most preferred embodiment the construct comprises an anti-Trop-2 antibody conjugated to Rap, although other combinations of antibodies, antibody fragments and toxins may be used to form the subject immunotoxins. The immunotoxins are of use to treat a variety of diseases, such as cancer, autoimmune disease or immune dysfunction.

Abstract: The invention concerns RAC-PK and fragments thereof, as well as activators and inhibitors of RAC-PK for use as medicaments, particularly in the treatment of diseases concerned with abnormalities in processes modulated by insulin, such as cellular proliferation, insulin deficiency and/or excess blood sugar levels. Moreover, the invention provides RAC-PK for use in screening potential mimics or modulators thereof. A method for screening for agents capable of affecting the activity of GSK3 is also disclosed. The invention further provides a screening kit comprising the RAC-PK as an active principle, and a method for screening compounds which are candidate mimics or modulators of RAC-PK activity comprising detecting specific interactions between the candidate compounds and RAC-PK. There is also provided a process for activating RAC-PK comprising treatment thereof with a phosphatase inhibitor.

Abstract: Isolated and/or purified polypeptides and nucleic acid sequences encoding polypeptides from Alicyclobacillus acidocaldarius are provided. Further provided are methods for modulating or altering recombination inside or outside of a cell using isolated and/or purified polypeptides and/or nucleic acid sequences from Alicyclobacillus acidocaldarius.

Abstract: The present invention, in some aspects, relates to systems and methods for determining oxidized proteins, including nitrosylated proteins such as S-nitrosylated proteins. The systems and methods of the invention can be used in vitro (e.g., in cell or tissue culture) or in vivo. For instance, in some cases, the invention can be used to spatially determine the location and/or concentration of oxidized proteins within cells and/or tissues (e.g., through visual detection). In one set of embodiments, a nitrosylated or otherwise oxidized moiety on a protein may be reacted with a detection entity, which may be, for example, fluorescent, radioactive, electron-dense, able to bind to a signaling entity or a binding partner in order to produce a signal, etc. In some embodiments, other moieties on the protein may be altered or blocked before reaction of the protein with the detection entity.

Abstract: The present invention relates to a recombinant microorganism capable of metabolizing sucrose, and more particularly to a recombinant microorganism capable of metabolizing sucrose in which a gene encoding sucrose phosphotransferase and/or a gene encoding sucrose-6-phosphate hydrolase is introduced or to a recombinant microorganism capable of metabolizing sucrose in which a gene encoding ?-fructofuranosidase is introduced. According to the present invention, a recombinant microorganism capable of using inexpensive sucrose as a carbon source instead of expensive glucose is provided. In addition, in a process of culturing microorganisms which have been incapable of using sucrose as a carbon source, sucrose can substitute for other carbon sources including glucose.

Abstract: The present invention provides, among other things, modified DNA polymerases containing amino acid alterations based on mutations identified in directed evolution experiments designed to select enzymes that are better suited for applications in recombinant DNA technologies.

Abstract: The present invention relates to Pregnane X receptor (PXR) polypeptides and crystals that are useful, for example, for crystallization and in assays for identification of modulators of PXR.

Abstract: Methods and compositions for increased production of amino acids from C. glutamicum using sucrose as a carbon source are described. In one aspect, increased production of L-lysine from C. glutamicum is accomplished by using a strain having a mutation in the ptsF gene encoding fructose-PTS enzyme that attenuates or blocks fructose import into the cell when such strain is grown on media containing sucrose as a carbon source and production is increased by providing glucose isomerase in the fermentation media. The glucose isomerase may be exogenously added or expressed in the strain and exported into the media. In certain embodiments the media also contain an invertase. In another aspect increased production of L-lysine is accomplished by making a C. glutamicum strain having the ptsF mutation and a second mutation in a fructose exporter function. The dual mutation retains imported fructose in the cell.

Abstract: The present invention provides a high growth reassortant influenza A virus having at least two gene segments of seasonal or pandemic strain origin, a PB1 gene segment of A/Texas/1/77 strain origin and a PA gene segment of A/Puerto Rico/8/34 (H1N1) origin coding for a PA protein comprising at least one amino acid modification at any one of positions 10, 275, 682, according to SEQ ID No.1. Further provided are vaccine formulations comprising the reassortant influenza A virus of the invention.

Abstract: The present invention relates to vectors for transforming archaea and to transformed archaea, and in particular to shuttle vector systems for transformation of members of the genus Pyrococcus.

Abstract: The present invention provides methods of detecting cancer using biomarkers.

Abstract: The invention relates to methods for amplifying, modifying, mutating and cloning DNA of any size. These methods comprise a series of PCR reactions, which are punctuated by ligation reactions.

Abstract: The invention is directed to methods for in vitro DNA synthesis catalysed by a DNA polymerase using cyclodextrins. The invention also relates to methods, compositions and kits comprising cyclodextrins for the amplification of a nucleic acid. The use of cyclodextrins improves the specificity, sensibility and/or yield of the amplification reaction. The invention is related more particularly to kits, compositions and methods for carrying out PCR reactions comprising a cyclodextrin.

Abstract: This invention relates generally to a method for producing a transgenic cell with increased gamma-aminobutyric acid (GABA) content as compared to a corresponding non-transformed wild type cell.

Abstract: The present invention relates to systems and methods for amplifying nucleic acid. In particular, systems and methods are provided for inhibiting polymerase based terminal transferase activity within a polynucleotide amplification setting (e.g., polymerase chain reaction). In addition, systems and methods are provided for generating amplified products generated with polynucleotide amplification techniques having reduced 3? non-templated nucleotide addition.

Abstract: The present invention relates to a formulation of a thermostable DNA polymerase which is completely free of detergents and its particular use in real time polymerase chain reaction (PCR). Such a formulation may be obtained if the selected purification method does not require the addition of a detergent at any purification step.

Abstract: The present invention relates to mutant DNA polymerases which incorporate dideoxynucleotides with about the same efficiency as deoxynucleotides. The present invention also related to mutant DNA polymerases which also have substantially reduced 5?-to-3? exonuclease activity or 3?-to-5? exonuclease activity. The invention also relates to DNA molecules coding for the mutant DNA polymerases, and hosts containing the DNA molecules.

Abstract: The present disclosure provides synthetic antibodies specific for a disease-associated antigen, and methods of using the antibodies in disease therapy. The present disclosure further provides diagnostic assays involving detecting the presence and/or level in biological sample of an antibody specific for a disease-associated antigen.

Abstract: The present invention relates to the cloning and characterization of a prokaryotic DNA repair ligase, which is shown to possess a range of activities that allow the ligation and repair of non-compatible DNA ends and double strand breaks (DSBs). The enzyme has a range of applications in the manipulation and cloning of nucleic acids.

Abstract: The present invention provides engineered novel variants of human immunodeficiency virus reverse transcriptase (HIV-RT) capable of being expressed in large quantity and that with polymerase and RNase H activity in a form that facilitates crystallization and high resolution structure resolution following X-ray diffraction. The present invention facilitates high resolution determination of RT in complexes with RT drugs and RT inhibitors, and provides methods for systematic generation of variants and for structure based identification and design of novel RT inhibitors.

Abstract: The present invention relates to mutant DNA polymerases that exhibit reduced discrimination against labeled nucleotides into polynucleotides. The DNA polymerases of the invention have at least one mutation in the nucleotide label interaction region of the enzyme such the mutation results in reduced discrimination against labeled nucleotides. The nucleotide label interaction regions is located at portions of the O-helix, (ii) the K helix, and (iii) the inter O-P helical loop of Taq DNA polymerase or analogous positions in other DNA polymerases. In addition to providing novel mutant DNA polymerases, the invention also provides polynucleotides encoding the subject mutant DNA polymerases. The polynucleotides provided may comprise expression vectors for the recombinant production of the mutant polymerases. The invention also provides host cells containing the subject polynucleotides.

Abstract: The use of raffinose as a glass-forming agent for freeze-dried compositions intended for use in conducting chemical or biochemical reactions such as the PCR. Thus, for instance, there is provided a composition for carrying out a chemical or biochemical reaction, said composition being in a freeze-dried form and comprising (i) a set of reagents comprising at least some of the chemical or biochemical reagents necessary for conducting said chemical or biochemical reaction, and (ii) raffinose. Kits comprising these compositions and methods of using them form a further aspect of the invention.

Abstract: The present invention provides materials and methods for antagonizing the function of vascular endothelial growth factor receptors, platelet derived growth factor receptors and other receptors. Soluble binding constructs able to bind vascular endothelial growth factors, platelet derived growth factors, and other ligands are provided.

Abstract: Modified amino acid sequences of OAS1 proteins in non-human primates, and genes related thereto, are provided.

Abstract: The isolation and purification of two domains from a from a flavivirus is provided. Each domain can function independently. Moreover, one domain codes for a sequence that provide polymerase activity. A process for screening possible modulators of the polymerase activity of an isolated and purified polypeptide from flavivirus is also disclosed.

Abstract: The present invention relates to novel markers for diagnosing brain disease caused by brain injury and the use thereof. More particularly, the present invention relates to novel diagnostic markers for brain disease caused by brain injury, which are identified by administering an MDMA drug, a diagnostic composition for brain disease caused by brain injury, a kit, a microarray, and a method for diagnosing brain disease caused by brain injury using the same, and relates to a method for screening a material for preventing or treating brain disease and a composition for preventing or treating brain disease caused by brain injury including the material. It was found that the expression amount of the marker genes of the present invention in tissues or cells of a patient with brain injury was over-expressed or under-expressed compared with that in normal tissues or cells.

Abstract: Disclosed are mutant DNA polymerases having improved extension rates relative to a corresponding, unmodified polymerase. The mutant polymerases are useful in a variety of disclosed primer extension methods. Also disclosed are related compositions, including recombinant nucleic acids, vectors, and host cells, which are useful, e.g., for production of the mutant DNA polymerases.

Abstract: The invention provides GmNFR1?, GmNFR1?, GmNFR5? and GmNFR5? soybean nodulation factor receptor proteins, a receptor complex and encoding nucleic acids. Also provided are GmNFR1?, GmNFR1?, GmNFR5? and GmNFR5? promoters which may be useful for expressing autologous or heterologous sequences in plants such as soybean. Variant proteins and nucleic acids including RNA splice variants, mis-sense mutants and non-sense mutants are also described. Also provided are genetically-modified plants and methods of producing genetically-modified plants. Over-expression of soybean nodulation factor receptor proteins by genetically-modified plants may lead to enhanced and/or otherwise facilitated nodulation and/or nitrogen fixation. Genetically-modified plants with down-regulated nodulation factor receptor expression, such as by RNAi or antisense constructs, may exhibit inhibited, diminished or otherwise reduced nodulation and/or nitrogen fixation.

Abstract: The present inventors found that a fusion gene present in some cancer patients is an oncogene. The present invention relates to a polypeptide as a novel fusion protein, a polynucleotide encoding the polypeptide, a vector comprising the polynucleotide, a transformed cell comprising the vector, a method for detecting the fusion protein or polynucleotide, a method for screening a therapeutic agent for cancer, and a method for treating cancer that is shown to be positive for the fusion gene. Further, the present invention relates kit, primer set, and probe useful in the detection of cancer that is shown to be positive for the fusion gene.

Abstract: The present invention relates to novel nucleic acid and protein molecules and their use in cancer therapy and diagnosis.