Abstract: The current invention relates to the use of luminescent oxygen channeling immunoassay (“LOCI”) technology to monitor amplification reactions, especially polymerase chain reactions (“PCR”). More specifically, the current invention involves the use of LOCI to measure the kinetics of a PCR reaction in an all-in-one assay format in order to quantitatively and qualitatively detect a target polynucleotide.

Abstract: Methods and pharmaceutical compositions useful for modulating lipoprotein levels in vivo. The invention stems from the discovery that activity of the Lipolysis Stimulated Receptor (LSR) can be inhibited or enhanced by exogenous agents, including polypeptides.

Abstract: This invention provides a DNA polymerase that is a mutant form of a naturally occurring DNA polymerase, of which one or more amino acids in the active site are mutated. The DNA polymerase mutant of this invention is characterized by altered fidelity or altered enzymatic activity in comparison with the naturally occurring DNA polymerase. For example, the DNA polymerase mutant provides increased enzymatic activity, altered dNTP/rNTP specificity, or enhanced fidelity. In one aspect of the invention, the naturally occurring DNA polymerase comprises an amino acid sequence motif: AspTyrSerGlnIleGluLeuArg in the active site. In another aspect of the invention, the naturally occurring DNA polymerase comprises an amino acid sequence motif: LeuLeuVa1AlaLeuAspTyrSerGlnIleGluLeuArg in the active site.

Abstract: Ligation methods for manipulating nucleic acid stands and oligonucleotides utilizing hybridization arrays of immobilized oligonucleotides. The oligonucleotide arrays may be plain or sectioned, comprehensive or non-comprehensive. The immobilized oligonucleotides may in some cases be binary oligonucleotides having constant as well as variable segments. Some embodiments include amplification of ligated products.

Abstract: The present invention provides a high-throughput screen for use in assaying for enzyme inhibitors of a target organism, comprising: a microorganism host in which an endogenous enzyme encoding gene has been replaced by a functionally complementing enzyme encoding gene from the target organism. The present invention also provides a method of identifying an inhibitor of a target organism enzyme, comprising the step of assaying a test inhibitor using a high-throughput screen comprising a microorganism host in which an endogenous enzyme encoding gene has been replaced by a functionally complementing enzyme encoding gene from the target organism.

Abstract: The present invention provides a sensitive assay for objectively determining the genotype of cucurbit plants, particularly species of melon, with respect to resistance or susceptibility to Fusarium wilt infection. The assay of the present invention uses a polymerase chain reaction to amplify sample DNA using either an AM or FM oligonucleotide primer pair. The PCR product which results from either primer pair differs in size, depending upon whether the template DNA was obtained from a plant susceptible or resistant to Fusarium wilt, permitting easy and rapid identification of plant genotype.

Abstract: EPRG3Spt polypeptides and polynucleotides and methods for producing such polypeptides by recombinant techniques are disclosed. Also disclosed are methods for utilizing EPRG3Spt polypeptides and polynucleotides in therapy, and diagnostic assays for such.

Abstract: A method for simultaneous release and detection of nucleic acids from complex biological samples is described. The invention relates to the combined use of lysis buffers containing strong chaotropic agents such as guanidine thiocyanate to facilitate cell lysis and release of cellular nucleic acids and to the use of a novel type of bicyclic nucleotide analogues, locked nucleic acid (LNA) to detect specific nucleic acids released during lysis by nucleic acid hybridisation. In particular methods are described for the covalent attachment of the catching LNA-oligo. Novel methods for sample preparation of e.g. polyadenylated mRNA species are also presented. The invention further addresses reagents for performing the methods as well as reagents and applications of the method.

Abstract: Modulating agents for inhibiting &bgr;-catenin mediated gene expression are provided. The modulating agents comprise one or more of: (1) the peptide sequence LXXLL (SEQ ID NO:1); or (2) a peptide analogue or peptidomimetic thereof. Methods for using such modulating agents for modulating &bgr;-catenin mediated gene expression and cellular differentiation in a variety of contexts (e.g., for modulating hair growth or treating cancer or Alzheimer's disease) are provided.

Abstract: A mammalian gene encoding a tolloid-like protein distinct from human or murine BMP-1/mTld is presented. The gene is similar in structure to members of the BMP-1 family of genes, but maps to a distinct location and encodes a distinct protein. The protein encoded by the gene can be used to screen putative therapeutic agents in an ongoing effort to inhibit activity of the BMP-1 family of genes to prevent scarring, fibrosis, and the like.

Abstract: The present invention relates to a method of diagnosing a mammal having abnormal prostatic cell growth or a predisposition to developing abnormal prostatic cell growth, said method comprising screening for the modulation of the expression of follistatin protein or derivative, homolog, analog, mutant, variant or fragment thereof in said mammal. More particularly, the present invention contemplates a method of diagnosing prostate cancer or a predisposition to developing prostate cancer, said method comprising screening for the co-expression of different forms of follistatin protein.

Abstract: This invention relates to diagnostic methods based upon a particular genotype in the Tumor Necrosis Factor (TNF&agr;) gene, more specifically, GA or AA at the −238 site rather than the GG at this locus. More specifically, this invention relates to a method for diagnosis of increased risk of death in patients with community-acquired pneumonia (CAP) and diagnosing pre-disposition or susceptibility to increased risk of death in patients who develop CAP, by screening for the presence of this polymorphism. The invention also relates to compositions for screening for the polymorphism and improved treatment choices for patients having the polymorphism of the present invention. The invention also relates to screening assays and therapeutic and prophylactic methods.

Abstract: An apparatus for amplifying DNA fragments is formed by a support plate. A plurality of openings are formed on the upper surface of the support plate. A plurality of primers having different amplification probabilities are arranged in the plurality of openings in order of the amplification probabilities. A plurality of microorganisms contained in a microorganism flora are simultaneously amplified with all primers by a random PCR method, for obtaining an electrophoretic pattern amplified at the optimum amplification probability for each microorganism. The plurality of microorganisms contained in the microorganism flora can be discriminated by analyzing the electrophoretic pattern.

Abstract: Sensitive methods for identifying compounds having biological activity comprising combining living cells with two fluorescent membrane permeable ionic dyes having the same charge sign, the first of which has an emission spectrum which overlaps the excitation spectrum of the second fluorescent membrane penetrative dye. The fluorescence is then induced by illuminating the dyes at a wavelength corresponding to the excitation spectrum of the first fluorescent dye and emission is then registered at a wavelength corresponding to the emission spectrum of the second fluorescent dye (FRET). The change in the FRET is indicative of a modulation of cell membrane potential by the biologically active compounds.

Abstract: A method for determining the genotype of one or more individuals at a polymorphic locus employs amplification of a region of DNA, labeling of allele-specific extension primers containing tags, and hybridization of the products to an array of probes. The genotype is identified from the pattern of hybridization. The method can also be used to determine the frequency of different alleles in a population.

Abstract: There is used at least one probe array obtained by arraying particles having various probes, respectively, fixed thereon (probe particles) in a definite order in a holder. A plurality of capillaries or grooves packed with various kinds, respectively, of probe particles are arrayed in parallel, and one of particles contained in each capillary or groove is injected into another capillary or groove to produce a probe array in which the various kinds of probe particles are arrayed in a constant and definite order. Various fluorophore-labeled DNA's are measured at the same time by attaching various probes to particles, respectively, of different sizes. A probe array composed of various fixed DNA probes can easily be produced, and there can be provided a probe array for detecting various DNA's which is composed of various fixed arbitrary DNA probes.

Abstract: A DNA expression vector for positively selecting in-frame or out-of-reading-frame mutations in DNA sequences to be tested comprising a promotor operatively linked to an expressible reporter gene through a linkage sequence is disclosed. The linkage sequence includes at least two restriction sites and an engineered frameshift mutation. In an embodiment the frameshift is established by complementary sequences SEQ ID Nos:1 and 2. The expressible reporter gene is expressed as a fusion product including a green fluorescent protein and the promoter can be lacZ and inducible in E. coli.

Abstract: A composition for the transfection of higher eucaryotic cells, comprising complexes of nucleic acid, a substance having an affinity for nucleic acid and optionally an internalizing factor, contains an endosomolytic agent, e.g. a virus or virus component, which may be conjugated. The endosomolytic agent, which is optionally part of the nucleic acid complex, is internalized into the cells together with the complex and releases the contents of the endosomes into the cytoplasm, thereby increasing the gene transfer capacity. Pharmaceutical preparations, transfection kits and methods for introducing nucleic acid into higher eucaryotic cells by treating the cells with the composition are also disclosed.

Abstract: The present invention relates to methods and kits for amplification of mRNA using a primer in PCR that contains an RNA polymerase promoter. The invention provides methods for amplification and detection of RNA derived from a population of cells, preferably eukaryotic cells and most preferably mammalian cells, which methods preserve fidelity with respect to sequence and transcript representation, and additionally enable amplification of extremely small amounts of mRNA, such as might be obtained from 106 mammalian cells. In typical embodiments of the invention, an RNA polymerase promoter (RNAP) is incorporated into ds cDNA by priming cDNA amplification by polymerase chain reaction (PCR) with an RNAP-containing primer. Following less than 20 cycles of PCR, the resultant RNAP-containing ds cDNA is transcribed into RNA using an RNA polymerase capable of binding to the RNAP introduced during cDNA synthesis.

Abstract: A screening test to identify women carrying a lethal genetic trait that predisposes to recurrent spontaneous pregnancy loss. The test method involves the quantitative determination of the frequency of highly skewed X chromosome inactivation in DNA derived from tissue cells of female patients, relative to appropriate normal control women. “Highly skewed” is defined as preferential use of one chromosome in at least 90% of the patient's cells being tested. Suitable test tissues include, but are not limited to, peripheral leukocytes, oral mucosal cells, and biopsy material.