Abstract: The entire process of reverse transcription-polymerase chain reaction (RT-PCR) is simplified by using oligonucleotide-immobilized microplates made of, e.g., polypropylene, to which oligonucleotides are securely immobilized and which can be subjected to thermal cycles of PCR. RT-PCR is preferably conducted in solid-phase. Capturing of mRNA and RT-PCR can be conducted in the same plates. The cDNA synthesized from the mRNA captured on the microplates can be used more than once. Further, in combination with the microplates, a filter plate is used for the preparation of cell lysates wherein target cells are placed on the filter plate, and a lysis buffer is passed through the cell layer on the filter to transfer cell lysate directly to the microplate via well-to-well communication.

Abstract: The invention provides a method for reducing stutter in the amplification of a microsatellite comprising the steps of providing a sample comprising a microsatellite having a G+C content of 50% or less; contacting the sample with at least one enzyme having nucleic acid polymerase activity; and incubating the sample with the enzyme for a sufficient amount of time and under conditions sufficient to amplify the microsatellite; wherein the incubation is performed in the presence of an amount of betaine, sorbitol or mixtures thereof, effective to reduce stutter relative to the amount of stutter observed in the absence of betaine and/or sorbitol. The invention also provides compositions containing betaine and/or sorbitol, kits for amplifying microsatellites having a G+C content of 50% or less, and methods of using all of the foregoing.

Abstract: Blood or other body fluid is screened for infection of an individual with HTLV-I and/or HTLV-II by subjecting each sample from the individual to a test for the presence of the Tax protein, DNA which encodes the Tax protein, or antibodies specific to the Tax protein, and correlating the presence of HTLV-I and/or HTLV-II infection with the result of the test. This test can also be used to screen pregnant women and nursing mothers for HTLV-I/II infection, or to screen seronegative patients who otherwise present symptoms of HTLV-I/II infection for HTLV-I/II infection. Because this test, which relies on testing for the presence of the tax protein is so specific for HTLV-I and/or HTLV-II infection, there is no requirement for input from any other test result to test positively for HTLV-I and/or HTLV-II.

Abstract: The invention relates to novel nucleic acid reference standards comprising a nucleic acid comprising a known target sequence bound with a microparticulate binding agent where the binding agent includes liposomes, polyamines (e.g., nylon), siliceous compounds (e.g., silica gel, fumed silica, diatomaceous earth, glass particles, amine-modified silica, and the like), zeolites (e.g., low alumina zeolyte), polystyrene (e.g., amine-modified polystyrene, carboxy-polystyrene particles, and the like), chitin, chitosan, and combinations of these compounds. The reference standard is useful for use as a standard in any nucleic acid assay where the presence or absence of a nucleic acid of interest is being assessed. The reference standard is stable and provides a control for assessing whether the nucleic acid assay was performed properly. The invention further relates to methods of producing such reference standards and kits for using and producing the same pursuant to the teachings of the invention.

Abstract: The present invention discloses nucleic acid detector probes for specific detection and/or quantification of target nucleic acid sequences and detection and/or quantification methods using these probes. In the absence of target nucleic acid sequence, a first oligonucleotide and a third oligonucleotide are bound to each other in a conformation which brings two member of an interacting moiety pair (labels) into close spatial proximity. Cooperative binding of the first oligonucleotide and a second oligonucleotide to a target nucleic acid sequence causes displacement of the third oligonucleotide from the first oligonucleotide probe resulting in separation of the two members of the interacting moiety pair (labels). The spatial separation of the moieties (labels) is detectable, and indicates the presence and/or amount of the target nucleic acid sequence.

Abstract: The invention relates to primer-specific and mispair extension assays for identifying gene variations, such as in different genotypes or subtypes of a given genotype. The assay includes extending a nucleic acid sequence from a patient sample with extension products of the primer, characterizing the extension products, and comparing the extension products with known nucleic acid sequences of various genotypes for determining the genotype of the nucleic acid sequence extended. In the assay, at least one primer or the dNTPs is labeled.

Abstract: The present invention relates to compositions and methods for releasing nucleic acid molecules from solid matrices. The invention further relates to compositions and methods for purifying and isolating nucleic acid molecules from biological materials such as animal tissues and plant matter. The methods of the invention can be readily adapted for rapid processing of multiple samples. Thus, the invention further provides automated methods for the purification of nucleic acid molecules from numerous samples. The invention also relates to kits for removing nucleic acid molecules from solid matrices.

Abstract: A pharmaceutical composition is disclosed, as well as its use in tumour diagnosis, therapy and prevention, methods for diagnosing, treating and preventing tumours, and antibodies and their use.

Abstract: A composite product is disclosed as a collagen support comprising at least one porous collagen layer covered on at least one side with an essentially compact collagen membrane consisting either of a collagen film prepared by drying a collagen gel, preferably in air or a gaseous fluid, or of a very highly compressed collagen sponge. At least one of the two layers, i.e. the porous layer and the essentially compact membrane, may comprise normal, genetically modified or malignant living cells originating particularly from young or elderly subjects. This composite product is used as a collagen support for the manufacture of artificial skin intended especially for performing in vitro tests on the efficacy of potentially active substances or for reconstructing damaged areas of skin in vivo.

Abstract: The invention relates to methods for cloning DNA molecules using recE/recT-mediated homologous recombination mechanism between at least two DNA molecules where one DNA molecule is a circular or linear DNA molecule and the second DNA molecule is a circular DNA molecule, and the second DNA molecule contains two regions with sequence homology to the first DNA molecule. Competent cells and vectors are also described.

Abstract: The invention provides promoters from Douglas-fir genes encoding metallothionein-like proteins. Also provided are deletions and variants of such promoters. The promoters are useful for, among other things, directing developmental-specific expression of transgenes.

Abstract: The present invention relates to method of identifying gene sequences of potential vaccine antigens. Also included are gene sequences and the polypeptides encoded by the gene sequences as well as the use of such sequences to induce a protective immune response in animals. Particularly, the invention relates to identifying potential antigen gene sequences of Mycoplasma, preferably Mycoplasma hyopneumoniae. In one aspect of the present invention there is provided a method of identifying expression proteins translated from a nucleotide sequence in an expression vector, said method comprising the use of a marker co-expressed with a protein translated from the nucleotide sequence. In a further aspect of the present invention there is provided a method of identifying a therapeutic antigenic gene sequence encoding a therapeutic antigenic protein of a disease, from a sample of nucleotide sequences. Preferably the marker is a polyHis tag.

Abstract: Screening methods for identifying compounds that regulate skeletal muscle atrophy or hypertrophy, regulate the activity or expression of the vasoactive intestinal peptide receptors (VPAC) or regulate expression of vasoactive intestinal peptide (VIP) or VIP analogs are provided. Methods for the prophylactic or therapeutic treatment of skeletal muscle atrophy utilizing VPAC as the target for intervention are described.

Abstract: The invention features methods of high throughput screening of candidate drug agents and rapid identification of drug targets by examining induction of the stress response in a host cell, e.g., the stress response in wildtype host cells and/or in host cells that differ in target gene product dosage (e.g., host cells that have two copies of a drug target gene product-encoding sequence relative to one copy). In general, induction of the stress response in wildtype host cells indicates that a candidate agent has activity of the drug. Induction of a relatively lower or undetectable stress response in a host cell comprising an alteration in gene product dosage indicates that the host cell is drug-sensitive and is altered in a gene product that plays a role in resistance to the drug.

Abstract: The present invention relates to the observation that women having an A→T 1890 polymorphism in the 3′ UTR of the cholinergic muscarinic receptor 2 (CHRM2) gene have an increased risk for developing major depression. The present invention provides diagnostic, screening and therapeutic methods based on that observation.

Abstract: The invention relates to methods and systems (e.g., computer systems and computer program products) for identifying and characterizing genes using microarrays. In particular, the invention provides for improved, robust methods for detecting genes through the use of microarrays to analyze the expression state of the genome. Genes which are expressed can be mapped to their respective positions in the genome, and the structure of such genes can be determined.

Abstract: This invention relates to a method of controlling sprout formation in plants and parts thereof including vegetative storage organs. The method involves the use of target and organ specific promoters to control expression of DNA sequences to inhibit sprouting. Sprouting is restored by switching on expression of DNA sequences using inducible promoter regions where sprouting may be controlled by, for example, application of an external chemical stimulus.

Abstract: Methods for isothermal exponential amplification of a target polynucleotide are disclosed. The methods employ two transcription modules, the first module providing linear amplification resulting in RNA transcripts, and a second module providing for further (generally cyclical) amplification resulting in more RNA transcripts. In one aspect, the amplification of the first module is composite primer based. In a second aspect, the amplification of the first module is based on target switching to generate a primer extension product comprising a promoter sequence. In all aspects, the RNA transcripts of the first transcription module are subjected to further amplification by creating an intermediate product comprising a double stranded promoter region from which transcription can occur. The invention further provides compositions and kits for practicing said methods, as well as methods which use the amplification results.

Abstract: The present invention provides methods and compositions for determining the presence and/or amount of Borrelia burgdorferi nucleic acids in a test sample related to Lyme disease. In particular, substantially purified oligonucleotide primers and probes are described that can be used for qualitatively and quantitatively detecting Borrelia burgdorferi nucleic acid in a test sample by amplification methods. The present invention also provides primers and probes for generating and detecting control nucleic acid sequences that provide a convenient method for assessing internal quality control of the Borrelia burgdorferi assay.

Abstract: Microcapsules prepared by encapsulating an aqueous solution of a protein, drug or other bioactive substance inside a semi-permeable membrane by are disclosed. The microcapsules are formed by interfacial coacervation under conditions where the shear forces are limited to 0-100 dynes/cm2 at the interface. By placing the microcapsules in a high osmotic dewatering solution, the protein solution is gradually made saturated and then supersaturated, and the controlled nucleation and crystallization of the protein is achieved. The crystal-filled microcapsules prepared by this method can be conveniently harvested and stored while keeping the encapsulated crystals in essentially pristine condition due to the rugged, protective membrane. Because the membrane components themselves are x-ray transparent, large crystal-containing microcapsules can be individually selected, mounted in x-ray capillary tubes and subjected to high energy x-ray diffraction studies to determine the 3-D structure of the protein molecules.