Abstract: Novel synthetic oligomers of unique three-dimensional structure comprising homologous and heterologous blocks, some of which may self-associate are described. Also disclosed is a synthetic oligonucleotide that includes 5-fluorocytosine residues at positions corresponding to methylation sites of DNA-cytosine methyltransferase and a method for preparing such a molecule.
Abstract: Methodology is provided for developing probes for identifying sequence differences between two related DNA populations, sets of DNA fragments or collections of restriction-endonuclease-cleaved DNA or cDNA. The method employs an initial stage to obtain a representation of both DNA populations, namely using the PCR to produce relatively short fragments, referred to as amplicons. Tester amplicons containing target DNA, sequences of interest, are ligated to adaptors and mixed with excess driver amplicons under melting and annealing conditions, followed by PCR amplification. The process may be repeated so as to greatly enrich the target DNA. Optionally, the target DNA may then be cloned and the DNA used as probes.
Abstract: A method for visualizing nucleic acids in a polyacrylamide gel. The method comprises fixing the nucleic acids with 10% acetic acid for about 20 minutes, washing the gel multiple times with water for about 2 minutes, impregnating the gel with silver nitrate at a concentration of about 1.0 g/l and about 1.5 ml/l of 37% formaldehyde for about 30 minutes, developing the gel with sodium carbonate at a concentration of about 30 g/l, 1.5 ml/l of 37% formaldehyde and sodium thiosulfate pentahydrate at a concentration of about 2.0 mg/l for between about 2 minutes and about 5 minutes, and then stopping the development of the gel by treatment with 10% acetic acid for about 5 minutes.
Type:
Grant
Filed:
February 8, 1994
Date of Patent:
February 20, 1996
Assignee:
University of Tennessee Research Corporation
Inventors:
Gustavo Caetano-Anolles, Brant J. Bassam, Peter M. Gresshoff
Abstract: A gene which is associated with tumor suppression and is localized on chromosome 11 has now been identified. The identification, localization and sequence of a gene which demonstrates differential expression in a manner that correlates with tumorigenicity suggests that this gene could potentially be used for gene therapy in cancers deleted or altered in their expression of the gene. Furthermore, a gene which is localized on chromosome 11p15, with identified polymorphisms, could be used for analysis of tumor DNA for loss of heterozygosity at chromosome 11p15. This region of chromosome 11 shows frequent loss of heterozygosity (LOH) in many human malignancies. Thus, the determination of LOH at chromosome 11p15 may be useful in predicting the prognosis of that tumor.
Type:
Grant
Filed:
January 5, 1995
Date of Patent:
February 13, 1996
Assignee:
The United States of America as represented by the Department of Health and Human Services
Abstract: Short nucleotide sequences of human papilloma virus useful for the determination of the presence and type of human papilloma virus present in a test sample. The sequences provided can be amplified by polymerase chain reaction or ligase chain reaction. The sequences provided also can be hybridized by standard slot-, dot- or replica-blot procedures. Methods and kits also are provided for the detection of human papilloma virus in a test sample and the determination of the type of human papilloma virus present in the test sample.
Type:
Grant
Filed:
September 30, 1994
Date of Patent:
January 16, 1996
Assignee:
Abbott Laboratories
Inventors:
Stanley R. Bouma, Jeffrey L. Joseph, Ronald L. Marshall, Thomas G. Laffler
Abstract: Products and methods particularly useful for activating and analyzing the nuclei and nucleic acids of human fetal red blood cells or cells found in amniotic fluid thereby facilitating prenatal screening are described. The featured products include activating egg extracts, cytostatic factor (CSF) extracts, kits containing these extracts, and a microchamber microscope slide useful in analyzing nucleus activation.
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.
Type:
Grant
Filed:
November 5, 1992
Date of Patent:
December 12, 1995
Assignees:
Institut Pasteur, Universite Paris-VI
Inventors:
Bernard Dujon, Andre Choulika, Laurence Colleaux, Cecile Fairhead, Arnaud Perrin, Anne Plessis, Agnes Thierry
Abstract: Process for the specific production of nucleic acids based on the principle of transcription in which a promoter oligonucleotide and a template-specific oligonucleotide which can hybridize with it are used as a promoter reagent and a process for nucleic acid detection which is based on this process.
Abstract: Compounds referred to herein as oligonucleotide clamps are provided that stably bind to target polynucleotides in a sequence-specific manner. The oligonucleotide clamps comprise one or more oligonucleotide moieties capable of specifically binding to a target polynucleotide and one or more pairs of binding moieties covalently linked to the oligonucleotide moieties. In accordance with the invention, upon annealing of the oligonucleotide moieties to the target polynucleotide, the binding moieties of a pair are brought into juxtaposition so that they form a stable covalent or non-covalent linkage or complex. The interaction of the binding moieties of the one or more pairs effectively clamps the specifically annealed oligonucleotide moieties to the target polynucleotide.
Abstract: A method that permits the rapid amplification of unknown DNA that flanks a known site, such that one can walk into an uncharacterized region of DNA. In this method, human genomic DNA is restriction enzyme digested and then ligated to a 5' phosphorylated-oligonucleotide so that the 5' end of each strand of genomic DNA is extended and phosphorylated. The phosphorylated-oligonucleotide is constructed to render 5' end extensions that are complementary to the known sequence. Following denaturation and re-annealing under dilute conditions that promote intrastrand annealing and under high stringency, only those DNA strands containing the known sequence will form a stem-loop structure with a recessed and phosphorylated 5' end, rendering a substrate for a subsequent heat-stable ligation reaction to another oligonucleotide. This second oligonucleotide is complementary to the sequence immediately adjacent to the phosphorylated-oligonucleotide high stringency annealing site.
Abstract: Oligonucleotides having substantially specific binding affinity towards C. difficile DNA are disclosed, especially oligonucleotides specific to the C. difficile A-toxin gene. Such oligonucleotides are useful as DNA probes and PCR primers in the detection of C. difficile in human clinical samples, e.g. fecal samples.
Type:
Grant
Filed:
May 19, 1992
Date of Patent:
October 17, 1995
Assignee:
3i Research Exploitation Limited
Inventors:
Soad Tabaqchali, Christopher L. Clayton, Brendan W. Wren
Abstract: Methods employing internal oligonucleotide standards in isothermal nucleic acid amplification reactions to determine the efficacy of the amplification reaction and to quantify pre-amplification target levels.
Abstract: Primers for amplification of specific nucleic acid sequences of the second and third exon of HLA Class I A gene and probes for identifying polymorphic sequences contained in the amplified DNA can be used in processes for typing homozygous or heterozygous samples from a variety of sources and for detecting allelic variants not distinguishable by serological methods. This HLA-A DNA typing system can be used in a forward or reverse dot-blot format that is simple and rapid to perform, produces detectable signals in minutes, and can be used for tissue typing, determining individual identity, and identifying disease susceptible individuals.
Type:
Grant
Filed:
September 28, 1993
Date of Patent:
September 19, 1995
Assignee:
Hoffmann-La Roche Inc.
Inventors:
Raymond J. Apple, Teodorica L. Bugawan, Henry A. Erlich
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.
Type:
Grant
Filed:
April 7, 1993
Date of Patent:
May 30, 1995
Assignee:
The Johns Hopkins University
Inventors:
Marilee Burrell, David E. Hill, Kenneth W. Kinzler, Bert Vogelstein
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 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: Novel DNA probe sequences for detection of CMV in a sample in a solution phase sandwich hybridization assay are described. Amplified nucleic acid hybridization assays using the probes are exemplified.
Type:
Grant
Filed:
October 15, 1993
Date of Patent:
April 18, 1995
Assignee:
Chiron Corporation
Inventors:
Janice A. Kolberg, Lu-Ping Shen, Michael S. Urdea
Abstract: Isolated nucleic acid molecules capable of hybridizing to sequences of Candida albicans along with methods utilizing such probes for the detection of Candida albicans in clinical and other biological samples.
Type:
Grant
Filed:
December 17, 1991
Date of Patent:
April 11, 1995
Assignee:
E. R. Squibb & Sons, Inc.
Inventors:
Jessica A. Gorman, Catherine A. Bingham
Abstract: The present invention provides an isolated nucleic acid encoding an approximately 120-128 kilodalton antigen of Helicobacter pylori, or an antigenic fragment thereof, wherein the antigen is associated with peptic ulceration. The present invention also provides methods of detecting the presence of a Helicobacter pylori strain possessing the 120-128 kilodalton antigen in a subject, comprising the steps of contacting an antibody-containing sample from the subject with a detectable amount of the tagA antigen or antigenic fragment of the present invention and detecting the reaction of the antigen or fragment and the antibody. A mutant H. pylori not expressing a functional tagA antigen is also provided.
Type:
Grant
Filed:
April 26, 1993
Date of Patent:
April 4, 1995
Assignee:
Vanderbilt University
Inventors:
Timothy L. Cover, Murali K. R. Tummuru, Martin J. Blaser