Abstract: The present invention provides in situ hybridization probes which include a marker and a nucleic acid molecule able to hybridize exclusively with only one species of Encephalitozoon. The nucleic acid molecule may be, for example, complimentary to segment 878-896 of 16S rRNA of Encephalitozoon hellem spores. Specifically disclosed probes are those including the following nucleotides: (1) 5?-ACT CTC ACA CTC ACT TCA G-3? (Seq. I.D. No. 1) which is species specific for Encephalitozoon hellem, (2) 5?-CAG ACC ACT ATC TGC A-3? (Seq. I.D. No. 2) which is species specific for Encephalitozoon cuniculi and (3) 5?-GTT CTC CTG CCC GCT TCA G-3? (Seq. I.D. No. 3) which is species specific for Encephalitozoon intestinalis. The assay of the present invention utilizes a sample such as surface, ground or drinking water, suspected of containing one of the aforementioned species as a target organism.

Abstract: The methods, compositions and apparatus disclosed herein are of use for nucleic acid sequence determination. The methods involve isolation of one or more nucleic acid template molecules and polymerization of a nascent complementary strand of nucleic acid, using a DNA or RNA polymerase or similar synthetic reagent. As the nascent strand is extended one nucleotide at a time, the disappearance of nucleotide precursors from solution is monitored by Raman spectroscopy or FRET. The nucleic acid sequence of the nascent strand, and the complementary sequence of the template strand, may be determined by tracking the order of incorporation of nucleotide precursors during the polymerization reaction. Certain embodiments concern apparatus comprising a reaction chamber and detection unit, of use in practicing the claimed methods. The methods, compositions and apparatus are of use in sequencing very long nucleic acid templates in a single sequencing reaction.

Abstract: The present invention provides a simple and quick method for determining the nucleotide sequence of the mitochondrial 21S ribosomal RNA gene of Saccharomyces cerevisiae by gene amplification technique, and a method for classifying Saccharomyces cerevisiae strains using the same nucleotide sequence. To achieve this object, the mitochondrial 21S ribosomal RNA gene of Saccharomyces cerevisiae is amplified by gene amplification technique, thereby determining the nucleotide sequence of the gene.

Abstract: The invention concerns single-chain antibodies directed against the p53 protein, capable of being expressed in tumoral cells, capable of restoring a DNA binding in vitro and a transcription activator function in vivo. The invention also concerns nucleic acids coding for these molecules, the vectors containing them and their uses.

Abstract: An objective of this invention is to provide a method for detecting DNA polymorphism that has high sensitivity and efficiency and does not need long DNA searching region. A homologous recombination protein RecA makes partial triple strand DNA from target double DNA and oligonucleotide probe complementary to the DNA. The triple strand DNA maintains stable triple strand DNA after RecA protein is removed. The present inventors found that the thermostability of triple strand DNA changes greatly when there is a mismatch between target DNA and oligonucleotide probe because of the existence of polymorphism in the target DNA. Utilizing this change of thermostability, efficient detection of polymorphism in labeled DNA is possible by examining whether oligonucleotide probe is released and the triple strand DNA is solved after heat treatment of triple strand DNA formed using homologous recombination protein.

Abstract: The present invention provides kits and methods for screening drugs and drug candidates for activity by determining the presence or absence of high integrity nucleic acid in a sample.

Abstract: A nucleic acid sequence comprising: 1) the sequence represented in FIG. 8; or 2) the sequence represented in FIG. 2; or 3) a part of the sequence of FIG. 2 with the proviso that it is able to code for a protein having a calcium dependant protease activity involved in a LGMD2; or 4) a sequence derived from a sequence defined in 1), 2) or 3) by substitution, deletion or addition of one or more nucleotides with the proviso that said sequences still codes for said protease.

Abstract: A method for rapidly identifying porcine estrogen receptor (ESR) marker comprises using published primers to amplify the target DNA fragment by polymerase chain reaction (PCR). The DNA fragment is cloned and then sequenced. The key positions of the sequence are modified to generate three primers which are used to amplify different DNA fragments with different genotypes by PCR to eliminate extra restriction cut reaction. A long one of the primers is to specifically amplify non-prolific allele, a short one of the primers is to specifically amplify prolific allele, and the remaining one is mutual and complimentary to the sequence. After PCR and electrophoresis, the sample with 90 bp band is identified as prolific genotype, the sample with 110 bp band is identified as non-prolific genotype, and the sample with 90 bp and 110 bp bands is identified as hetero-genotype.

Abstract: Methods, compositions, kits, and apparatus are provided wherein the aminoacyl-tRNA synthetase system is used to analyze amino acids. The method allows very small devices for quantitative or semi-quantitative analysis of the amino acids in samples or in sequential or complete proteolytic digestions. The methods can be readily applied to the detection and/or quantitation of one or more primary amino acids by using cognate aminoacyl-tRNA synthetase and cognate tRNA. The basis of the method is that each of the 20 synthetases and/or a tRNA specific for a different amino acid is separated spatially or differentially labeled. The reactions catalyzed by all 20 synthetases may be monitored simultaneously, or nearly simultaneously, or in parallel. Each separately positioned synthetase or tRNA will signal its cognate amino acid. The synthetase reactions can be monitored using continuous spectroscopic assays.

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 relates to a method for detecting microorganisms in a sample by means of a nucleic acid probe. Conventional detection methods are, for example, the in-situ hybridization of microorganisms with fluorescence-labeled oligonucleotide probes (fluorescent in-situ hybridization). A disadvantage of said method is that an epifluorescence microscope is required for evaluating the results. According to the invention, the disadvantages of the in-situ hybridization method are overcome by hybridizing the microorganisms to be detected in a sample with a specific nucleic acid probe, removing non-hybridized nucleic acid probe molecules, separating and then detecting and optionally quantifying the hybridized nucleic acid probe molecules.

Abstract: The present invention relates to a method for diagnosing panic disorder, said method being based on the determination of the class of polymorphism of the short tandem repeat (STR) complex in the upstream region of human cholecystokinin gene. The polymorphic DNA comprises a DNA sequence, having a general formula (1): 5?(GGAA)n1X(GGAG)n2(GGAA)n3(GGGA)n4GAG(AGAC)n5Y(GGAA)n63???(1) wherein X denotes a DNA sequence of SEQ ID NO: 1, Y denotes a DNA sequence of SEQ ID NO:2 and each of n1, n2, n3, n4, n5 and n6 denotes independently 0 or a positive integral number, whereby said DNA ranging from 363 to 399 base pairs in length. The invention also relates to an assay kit used for implementing said method.

Abstract: Methods and kits are disclosed for use in the analysis of microsatellite instability in genomic DNA. Methods and kits are also disclosed which can be used to detect microsatellite instability DNA present in biological materials, such as tumors. The methods and kits of the present invention can be used to detect or diagnose diseases associated with microsatellite instability, such as certain types of cancer.

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: isolated coding sequences and to the protein sequences they code for. This invention is directed to three coding sequence of the BRCA1 gene. The three coding sequences, BRCA1(omi1), BRCA1(omi2), and BRCA1(omi3) and their frequencies of occurrence are provided together with the protein sequences they code for. Another aspect of this invention is a method of determining the consensus sequence for any gene. Another aspect of the invention is a method of identifying an individual having an increased genetic susceptibility to breast or ovarian cancer because they have inherited a causative mutation in their BRCA1 gene. This invention is also related to a method of performing gene therapy with any of the isolated BRCA1 coding sequences.

Abstract: Novel non-coding sequences isolated upstream of the human IRS-2 gene are disclosed as markers for the prediction and/or diagnosis of IRS-2 related metabolic disorders or diseases, such as diabetes. The sequences also fuction as markers in a method and assay for evaluating the insulin regulating, i.e. insulin sensitizing or inhibiting properties of drug candidate substances, e.g. a method and assay for high throughput screening. The sequences and/or information derived therefrom can also be used for influencing the expression of the IRS-2 gene, e.g. in the therapy of IRS-2 related metabolic disorders, such as diabetes.

Abstract: An isolated DNA encoding the enzyme I-SceI is provided. The DNA sequence can be incorporated in cloning and expression vectors, transformed cell lines and transgenic animals. The vectors are useful in gene mapping and site-directed insertion of genes.

Abstract: The invention relates to the sequencing of a target polynucleotide sequence, immobilized on a solid support, using the polymerase reaction to extend a suitable primer and characterizing the sequential addition of labelled bases. The present invention further relates to the presence of a polymerase enzyme that retains a 3′ to 5′ exonuclease function, which is induced to remove an incorporated labelled base after detection of incorporation. A corresponding non-labelled base may then be incorporated into the complementary strand to allow further sequence determinations to be made. Repeating the procedure allows the sequence of the complement to be identified, and thereby the target sequence.

Abstract: The present invention provides a method, oligonucleotide probes and primers, and assay kits for diagnosing an individual as having hypospermatogenesis associated infertility that involves demonstrating in the individual the presence or absence of a variant form of a gene in exons 6, 7, 8 or 9 of the L-VDCC &agr;1c transcript.

Abstract: Novel amine compounds are provided by the present invention. Methods of preparing and using said novel amine compounds are also provided.