Abstract: A human gene has been discovered which is genetically altered in human tumor cells. The genetic alteration is gene amplification and leads to a corresponding increase in gene products. Detecting that the gene, designated hMDM2, has become amplified or detecting increased expression of gene products is diagnostic of tumorigenesis. Human MDM2 protein binds to human p53 and allows the cell to escape from p53-regulated growth.

Abstract: Xanthomonas campestris pv. citri is a quarantine organism under United States and International law because of the serious disease of citrus, citrus bacterial canker disease, which is caused by the organism. We have cloned, in vector pUC9, a 4.2-kb BamHI fragment of plasmid DNA from a typical strain of this pathogen and demonstrated that this DNA fragment specifically identifies the pathogen. The procedure involves isolation and cultivation of the bacterium, chemical isolation of its DNA, digestion of the DNA by restriction endonucleases and analysis by Southern or dot blotting using the cloned DNA fragment as biotin-labeled hybridization probe. A subclone has been made from the original 4.2-kb BamHI fragment which has sensitivity and specificity equal or greater than the original clone and which is approximately 572 bp in length. All tested strains of the most virulent form of the pathogen, type A, have a BamHI fragment of 4.2-kb which hybridizes with either probe.

Abstract: The present invention relates to a fat cell specific rat .beta.-adrenergic receptor that mediates lipolysis in rats. The invention further relates to cloned cells which code for the specific .beta.-adrenergic receptor that mediates lipolysis. Another aspect of the present invention relates to a diagnostic test method for determining decreased levels of fat cell .beta.-adrenergic receptors that mediate lipotysis in order to diagnosis obesity caused by less active lipolysis.

Abstract: Improved methods for amplifying nucleic acids can reduce non-specific amplification and minimize the effects of contamination of nucleic acid amplification reaction assays due to amplified product from previous amplifications. The methods involve the introduction of unconventional nucleotide bags into the amplification reaction products and treating the products by enzymatic (e.g., glycosylases) and/or physical-chemical means to render the product incapable of acting as a template for subsequent amplifications.

Abstract: A method for amplifying nucleic acid sequences in a template comprising at least one nucleic acid or mixture of nucleic acids wherein each nucleic acid comprises at least one strand. The method comprises the steps of treating the template with at least one oligonucleotide primer having a nucleic acid sequence that is substantially complementary to at least one nucleic acid sequence on the template under conditions such that the primer will bind to a segment that is substantially complementary to the primer, producing primed strands of nucleic acid. The primed strands produce extension strands, wherein the extension strands comprise the primer in combination with a sequence of nucleic acids that is substantially complementary to the template in a 5' to 3' direction from the primer. The extension strands are further treated with the primer producing amplification strands which are treated with primer.

Abstract: Method of determining polymerase activity by incubating the polymerase with a template nucleic acid, one detectable mononucleoside triphosphate and one immobilizable nucleoside triphosphate, binding the immobilizable nucleotides to a solid phase and detecting the bound detectable nucleotides and tests based thereon.

Abstract: A multifunctional reagent is provided that is useful for the attachment of desired molecules to support surfaces. A reactive reagent molecule of the invention is "restrained" in that it is conformationally and/or chemically restricted from reacting with either itself or with other molecules of the same reagent. Upon activation, this feature causes the attachment of less than all of the reactive sites of the multifunctional reagent to a surface, thereby leaving the remaining sites free to react with molecules desired to be immobilized onto the surface.

Abstract: A method for isolating a cDNA specific for the human ryanodine receptor is disclosed. The gene is associated with malignant hyperthermia, a hypermetabolic syndrome triggered primarily by inhalation anesthetics. The cDNA can be cloned and expressed in a recombinant plasmid or phage. The cDNA, or fragments thereof, is used as diagnostic probes for individuals at risk for malignant hyperthermia using restriction fragment length polymorphism analysis. The cDNA is that sequenced in FIG. 2 of this specification.

Abstract: Improved method for rapidly sequencing elements constituting a linear or linearized and ordered biological sequence such as a DNA fragment is provided. The method consists of extracting purifying and where appropriate fragmenting and/or amplifying a biological sequence in order to obtain a plurality of identical sequences which are subsequently combined to identify the complete DNA fragment.

Abstract: A novel rapid mutational analysis method for mapping protein epitopes is disclosed. This method has been used to identify the binding sites for 16 anti-CD2 and anti-CD4 monoclonal antibodies. The powerful, rapid, and simple method of the present invention allows isolation of a very large number of mutants, and is applicable to any intracellular or surface protein for which a cDNA and monoclonal antibodies are available. The present method is especially useful in ligand binding site studies for the design of new ligands and drugs.

Abstract: A human gene has been discovered which is genetically altered in human tumor cells. The genetic alteration is gene amplification and leads to a corresponding increase in gene products. Detecting that the gene, designated hMDM2, has become amplified or detecting increased expression of gene products is diagnostic of tumorigenesis. Human MDM2 protein binds to human p53 and appears to allow the cell to escape from p53-regulated growth.

Abstract: Improvements in the existing procedures and materials for conduct of high gradient magnetic separation (HGMS) are disclosed. Superior superparamagnetic particles, optionally coated with a polysaccharide or other, usually organic, materials can be prepared in uniform compositions with homogeneous magnetizations. The coating can conveniently be conjugated to a specific binding moiety complementary to a biological material whose purification or separation is desired. In addition, plastic coated matrices which form superior magnetic gradient-intensifying supports are disclosed, along with improved methods and apparatus to conduct HGMS.

Abstract: Improvements to the polymerase chain reaction (PCR), a process for in vitro enzymatic amplification of specific nucleic acid sequences, can be achieved by changing the way that PCR reagents are mixed and the enzymatic reaction is started and by the replacement of mineral oil, commonly used as a vapor barrier to minimize solvent evaporation, by a grease or wax. The use of such mixtures allows for the delay of reagent mixing until the first heating step of a PCR amplification, thereby reducing the enzymatic generation of nonspecific products which occurs when a complete mixture of PCR reagents, with or without test sample, stands at room temperature or below. These mixtures increase the shelf-life of PCR reagents and increase protection of the laboratory environment against contamination by PCR product.

Abstract: The present invention is related to the identification of cloned DNA sequences that reveal individual multiallele loci. The loci are used in the process of the present invention to provide convenient and accurate genetic identification. A large number of clones that recognize VNTR loci have been isolated from a cosmid library and characterized.

Abstract: A method that permits the highly specific PCR amplification of unknown DNA that flanks a known sequence region. In this method, known DNA is placed on the uncharacterized side of that specific sequence that contains the unknown DNA by a DNA polymerase mediated generation of a PCR template that is shaped like a pan with a handle. Generation of this template permits specific amplification of the segment of interest.

Abstract: Novel signal peptides capable of functioning in yeasts have the amino acid sequence:Met-A.sub.1 -A.sub.2 -X-B-C-D-E-Fwherein A.sub.1 is a peptide chain composed of 1-3 amino acids each selected from the group consisting of Arg, Ser, Lys and His, A.sub.2 is a peptide chain composed of 1-3 amino acids, X is a peptide chain composed of 8-10 hydrophobic amino acids, B is Pro or Ser, C is Gly or Pro, D Cys, Ala, Leu, Ser, Thr and Val, E is Trp or Gln and F is Ala or Gly. DNA sequences coding for these signal peptides can be used for the secretory expression of heterologous proteins in yeasts.

Abstract: The complete cDNA sequence encoding the amino acid sequence corresponding to mammalian Na.sup.+ /nucleoside cotransporter protein (SNST) is disclosed. Methods for obtaining the gene encoding SNST and for obtaining recombinantly produced SNST are described. Antibodies, an inhibitor of nucleoside transport by SNST, and methods for detecting other inhibitors are also described. Methods for inhibiting uptake of nucleosides by SNST using the compositions of the invention are also included.

Abstract: This invention relates to a methodology for assessing the sensitivity of an HIV-1 sample to zidovudine and to diagnostic assays for use in such assessment.

Abstract: The present invention relates to the identification of yeast strains by way of application of the polymerase chain reaction to amplify nucleic acid sequences characteristic of their TY transposon long terminal repeats. Polymerase chain reaction product is analysed, conveniently by agarose gel electrophoresis, and its nature related to the presence of a particular yeast strain or strain type.

Abstract: Methods and compositions are described for determining the level of low density lipoproteins (LDL) in plasma. Native apoprotein B-100 (apo B-100) present in LDL particles is immunologically mimicked by a polypeptide of the invention. A polypeptide includes an amino acid residue sequence corresponding to a pan epitope region of the target apoprotein. A preferred polypeptide is a fusion protein that simultaneously mimics native apo B-100 and native apo A-I. Improved assay systems and methods for determining HDL and LDL levels in a body fluid sample are also described.