Abstract: Methods and kits for the use of a non-ionic detergent to suppress enzyme inhibition in a reaction solution due to the presence of inhibiting ionic detergent. Prior to reaction, the reaction mixture is given an effective amount of a non-ionic detergent, and agitated. The enzyme is then added, and the enzymatic reaction is the allowed to proceed. Also disclosed are preferred embodiments of the present invention, and kits for nucleic acid amplification of a biological sample in the presence of an ionic detergent.

Abstract: Improved methods for determining interactions between peptides or proteins and small molecules are disclosed. The invention methods can be used to screen libraries of either the small molecules or the proteins. In general, the methods comprise contacting an agent/ligand complex consisting essentially of an agent to be tested for binding to a target protein coupled to a ligand capable of binding a proteinaceous ligand-binding domain with a first fusion protein comprising said target protein and a first complementary portion of a segregable protein; and a second fusion protein comprising a proteinaceous ligand-binding domain and a second complementary portion of said segregable protein; and detecting whether the first complementary portion and second complementary portion are brought into proximity.

Abstract: The present invention provides methods and apparatus for detecting and discriminating multiple analytes within a test sample which are simple, user-friendly, cost-effective and fast. In particular, it is preferred that the overall time for sample preparation, nucleic acid sequence amplification, and nucleic acid sequence differentiation be about 5 hours or less.

Abstract: The present invention relates to means for the detection and characterization of nucleic acid sequences, as well as variations in nucleic acid sequences. The present invention also relates to methods for forming a nucleic acid cleavage structure on a target sequence and cleaving the nucleic acid cleavage structure in a site-specific manner. The 5' nuclease activity of a variety of enzymes is used to cleave the target-dependent cleavage structure, thereby indicating the presence of specific nucleic acid sequences or specific variations thereof.

Abstract: This invention provides a method to normalize a cDNA library comprising: (a) constructing a directionally cloned library containing cDNA inserts wherein the insert is capable of being amplified by polymerase chain reaction; (b) converting a double-stranded cDNA library into single-stranded DNA circles; (c) generating single-stranded nucleic acid molecules complementary to the single-stranded DNA circles converted in step (b) by polymerase chain reaction with appropriate primers; (d) hybridizing the single-stranded DNA circles converted in step (b) with the complementary single-stranded nucleic acid molecules generated in step (c) to produce partial duplexes to an appropriate Cot; and (e) separating the unhybridized single-stranded DNA circles from the hybridized DNA circles, thereby generating a normalized cDNA library.

Abstract: The present invention concerns a novel DNA sequence coding for an Aspergillus serine protease of the subtilisin-type, an Aspergillus serine protease of the subtilisin-type per se and a method for the preparation thereof. The invention further concerns a novel Aspergillus mutant strain defective in a serine protease of the subtilisin-type, which is useful for the expression of heterologous protein, and a method for the preparation of such a mutant strain.

Abstract: Antisense constructs ditrected against viral cis-acting post-transcriptional regulatory sequences ("PREs") are disclosed. In one embodiment, the antisense construct is an expression plasmid encoding one or more antisense transcripts which hybridize under intracellular conditions to all or a portion of a viral PRE within a viral transcript, the PRE having the function of directing export of the viral transcript from the nucleus to the cytoplasm of a cell. The antisense constructs can be used to inhibit viral production, such as HBV production.

Abstract: The present invention relates to a disease-controlling agent for useful gramineous plants such as rices, corns, wheats, barleys, ryes and oats, comprising as an active ingredient a microorganism belonging to the genus Exserohilum and having an ability to control diseases of useful gramineous plants, as well as a disease control method for gramineous plants comprising applying said agent to useful gramineous plants including rices, corns, wheats, barleys, ryes and oats.

Abstract: The present invention relates to isolated nucleic acid constructs containing a sequence encoding a Scytalidium laccase, and the laccase proteins encoded thereby.

Abstract: A method for the detection of poor metabolizers of drugs using PCR technology is described comprising a) optionally amplifying a gene coding for an enzyme known to be responsible for the metabolization of drugs, thereby separating it from possible closely related pseudogens, b) amplifying different allelic forms of the gene of step a) and c) detecting the product of step b). Primers for amplification of the genes responsible for the debrisoquine or acetylation phenotype are also disclosed.

Abstract: A method is disclosed for isolating plasmid DNA, having the steps of: suspending hydroxyapatite particles in a buffer solution having a pH of about 6 to about 9, about 1 to about 100 mM of a water-soluble calcium salt and about 1 to about 100 mM of tris(hydroxymethyl) aminomethane; adding a solution containing RNA and plasmid DNA, obtained upon bacteriolysis, to the hydroxyapatite particles-containing suspension, thereby adsorbing the RNA onto the hydroxyapatite particles in the suspension; and recovering the plasmid DNA from a supernatant of the suspension. The isolation method enables effective separation of the purified plasmid DNA from a solution containing RNA and plasmid DNA in a simplified process, without the use for any special separation device.

Abstract: The present invention relates to means for cleaving a nucleic acid cleavage structure in a site-specific manner. Enzymes, including 5' nucleases and 3' exonucleases, are used to screen for known and unknown mutations, including single base changes, in the human p53 gene. Methods are provided which allow for the identification of genetic mutations in the human p53 gene in a sample.

Abstract: The present invention and kits are directed to a method of amplifying and detecting single or double-stranded target nucleic acid molecules in a test sample. Amplification is accomplished through the use of a minimum of two oligonucleotide probe complement pairs, wherein members oligonucleotide probes from both pair of oligonucleotide probe complement pairs form a minimum of two oligonucleotide probe pairs, at least one of which is complementary to a given portion of a target nucleic acid sequence which act as template. One of the oligonucleotide probes of each oligonucleotide probe pair have an additional protecting sequence which is not complementary to the target sequence. These additional protecting sequences are preferably complementary to each other. Chemical functionality groups attached to the oligonucleotide probes covalently combine the probes to form a joined oligonucleotide product. The joined oligonucleotide product is formed without the use of enzymes.

Abstract: The present invention features a method for introducing hypermutations into a target DNA or RNA sequence of interest, characterized in that said method comprises the steps of:(a) transcribing a RNA into DNA in a reaction mixture comprising a reverse transcriptase, varying biased concentrations of deoxynucleoside triphosphates to produce hypermutations and an oligonucleotide primer that is partially complementary to the 3' end of said RNA; and(b) recovering said DNA sequences.

Abstract: A nucleic acid comprising a base sequence which codes for a CEA peptide sequence or nucleic acids having a base sequence hybridizable therewith, replicable recombinant cloning vehicles having an insert comprising such nucleic acid, cells transfected, infected or infected with such cloning vehicles, polypeptides expressed by such cells, antibody preparations specific for such polypeptides, immunoassays for detecting CEA using such antibody preparations and nucleic acid hybridization methods for detecting a CEA nucleic acid sequence using a nucleic acid probe comprising the above described nucleic acid.

Abstract: A method and composition are disclosed for transfecting eukaryotic cells using a DNA segment coupled to a site-specific chromosome-binding polypeptide. The polypeptide-DNA conjugate is referred to herein as a polypeptide-linked-rDNA (PLR) molecule. One example of a PLR molecule comprises a DNA segment containing a nucleotide sequence from a normally functioning human gene, coupled by means of a covalent crosslinking reagent to a site-specific chromosome-binding polypeptide (such as a transcription regulating polypeptide that binds to a specific nucleotide sequence in chromosomal DNA). After the PLR molecule enters the cytoplasm of a cell, such as by electroporation, the chromosome-binding polypeptide enables transport of the PLR molecule through the cytoplasm and into the nucleus, using a nuclear localization sequence (NLS) domain of the polypeptide. Inside the nucleus, the polypeptide scans the chromosomes until it binds to a specific chromosomal binding site.

Abstract: A method for conferring resistance to a parasite to a host of the parasite, which comprises isolating a gene fragment from the parasite and inserting the gene fragment or a DNA or RNA segment substantially homologous to the gene fragment or to a DNA or RNA sequence functionally equivalent to the gene fragment into the host, wherein (1) transcription of the gene fragment or the DNA or RNA segment in the host occurs in an anti-sense direction, (2) the gene fragment or the DNA or RNA segment is expressed as a gene product in the host, wherein the gene product is capable of disrupting an essential activity of the parasite, or (3) the gene fragment or the DNA or RNA segment is a binding site capable of competing with a native binding site in the parasite, is disclosed along with hosts produced by this process. Particularly preferred is conferring resistance using a gene fragment from a replicase gene of an RNA virus.

Abstract: Methods for detecting hematologic and colorectal malignancies are provided comprising: (a) collecting a sample suspected of containing cancer cells; (b) analyzing the sample for telomerase activity; (c) correlating the presence of telomerase activity with the presence of cancer cells. A method for staging leukemia is also provided comprising analyzing a blood or bone marrow sample for telomerase activity, correlating the activity with a standard level of telomerase activity, and correlating a low telomerase activity with early stage leukemia.

Abstract: The invention relates to the identification of the molecular basis of supravalvular aortic stenosis (SVAS) and Williams syndrome. More specifically, the invention has identified that elastin causes or is involved in the pathogenesis of SVAS and Williams syndrome. Molecular variants of the elastin gene contribute to SVAS and Williams syndrome. The analysis of the elastin gene will provide an early diagnosis of subjects with SVAS and Williams syndrome. The diagnostic method comprises analyzing the DNA sequence of the elastin gene of an individual to be tested and comparing it with the DNA sequence of the native, non-variant elastin gene. In a second embodiment, the elastin gene of an individual to be tested is screened for mutations associated with SVAS or Williams syndrome. Presymptomatic diagnosis of SVAS and Williams syndrome will enable practitioners to prevent vascular obstruction using existing medical therapies like beta adrenergic blocking agents.

Abstract: Antisense oligonucleotides to nerve growth factor receptor, p75.sup.NGFR gene downregulate expression, thereby facilitating neurone survival.

Abstract: Modified Protein C molecules have been made which substitute the gamma carboxylglutamic acid (Gla) region of another Vitamin K dependent protein, most preferably prothrombin, for the native region of the Protein C. The modified or chimeric protein C has advantages over the wild-type protein C since it is less sensitive to inhibition by natural inhibitors of protein C (which would otherwise decrease the ability of the protein C to act as an anticoagulant) and which does not need the same cofactors or same amounts of cofactors, and can therefore be effective in patients with lowered levels of the cofactors such as protein S or the lipids present in activated platelets such as phosphatidyl ethanolamine (PE).

Abstract: The subject invention concerns the novel use of formyl-CoA transferase enzyme together with oxalyl-CoA decarboxylase enzyme for the detection and measurement of oxalate in biological samples. The use of the enzyme system according to the subject invention results in the conversion of oxalate into carbon dioxide and formate. Because the production of formate is directly correlated to the concentration of oxalate present in a sample, the determination of the resulting formate concentration provides an accurate, sensitive and rapid means for detecting even low levels of oxalate. The subject invention further concerns the cloning, sequencing and expression of the genes that encode the formyl-CoA transferase enzyme and the oxalyl-CoA decarboxylase enzyme of Oxalobacter formigenes. The subject invention also concerns a method for detecting the presence of Oxalobacter formigenes organisms in a sample, and the polynucleotide probes and primers used in the detection method.

Abstract: RNA is transcribed from a double stranded DNA template by forming a complex by hybridizing to the template at a desired transcription initiation site one or more oligonucleic acid analogues of the PNA type capable of forming a transcription initiation site with the DNA and exposing the complex to the action of a DNA dependant RNA polymerase in the presence of nucleoside triphosphates. Equal length transcripts may be obtained by placing a block to transcription downstream from the initiation site or by cutting the template at such a selected location. The initiation site is formed by displacement of one strand of the DNA locally by the PNA hybridization.

Abstract: DNA chips containing arrays of oligonucleotide probes can be used to determine whether a target nucleic acid has a nucleotide sequence identical to or different from a specific reference sequence. The array of probes comprises probes exactly complementary to the reference sequence, as well as probes that differ by one or more bases from the exactly complementary probes.

Abstract: The present invention is related to an improved method for the quantification of nucleic acid, which can be performed with a minimal amount of nucleic acid amplification reactions. The method according to the invention for the quantification of analyte nucleic acid in a sample comprises the steps of: adding to the sample different respective amounts of different nucleic acid constructs, each construct being distinguishable from the analyte nucleic acid and capable of being co-amplified with the analyte nucleic acid; subjecting the sample to a nucleic acid amplification procedure, using amplification reagents capable of reacting with both the analyte nucleic acid and the nucleic acid constructs; detecting the relative amounts of amplificates derived from analyte nucleic acid and each nucleic acid construct; calculating the amount of analyte nucleic acid from said relative amounts.

Abstract: A preventive or therapeutic agent for Alzheimer's disease which comprises a substance exhibiting an inhibitory action to tau-protein kinase I as an effective component is provided. A pharmaceutical composition comprising said agent and a method of inhibiting neuronal cell death in the brain are also provided.

Abstract: The present invention provides methods for identifying genetic alterations in a target sequence present in a nucleic acid sample, comprising immobilizing samples on a support, contacting the samples simultaneously with different purine and pyrimidine containing polymers under conditions of hybridization, separating the hybridized polymers from the samples; and identifying the hybridized polymers to identify the genetic alteration(s). The present invention also provides methods for identifying target sequences present in a nucleic acid sample, comprising immobilizing nucleic acid samples on a support, contacting the samples simultaneously different purine and pyrimidine containing polymers under hybridization conditions, separating the polymers from the complementary target sequence(s), and identifying the hybridized polymers to identify the target sequence(s).

Abstract: Methods for amplifying a nucleic acid molecule which employs a single primer, and in which the amplification is performed under isothermal conditions. The invention also includes kits containing reagents for conducting the method.

Abstract: Regulatable RNA molecules such as regulatable ribozymes, nucleic acids encoding such regulatable ribozymes, and methods of making and using such regulatable ribozymes are disclosed. Regulatable ribozymes comprise a ligand-binding RNA sequence and a ribozyme sequence capable of cleaving a separate targeted RNA sequence, wherein upon binding of the ligand to the ligand-binding RNA sequence, the activity of the ribozyme sequence against the targeted RNA sequence is altered. The ligand may be either an inorganic or an organic molecule and may be a co-drug which can be administered to specifically regulate the ribozyme activity. Regulatable RNA molecules other than ribozymes are also disclosed, such as regulatable mRNA molecules which comprise a ligand-binding RNA sequence separate from the coding sequence, wherein upon binding of a ligand to the ligand-binding RNA sequence, translation of the regulatable mRNA is altered.

Abstract: The allelic type of a polymorphic genetic locus in a sample is identified by first combining the sample with a sequencing reaction mixture containing a polymerase, nucleotide feedstocks, one type of chain terminating nucleotide and a sequencing primer to form a plurality of oligonucleotide fragments of differing lengths, and then evaluating the length of the oligonucleotide fragments. As in a standard sequencing procedure, the lengths of the fragments indicate the positions of the type of base corresponding to the chain terminating nucleotide in the extended primer. Instead of performing and evaluating four concurrent reactions, one for each type of chain terminating nucleotide, however, the sample is concurrently combined with at most three, and preferably only one, sequencing reaction mixtures containing different types of chain terminating nucleotides. The information obtained from this test is evaluated prior to performing any additional tests on the sample.

Abstract: The present invention relates to the gene for stilbene synthase, isolated from plants, and its use for the transformation of vectors, host organisms and plants, as well as to the production of plants which have an increased resistance to pests.

Abstract: The invention relates to DNA polymerases which are capable of proofreading 3'-5' exonuclease activity during DNA sequencing of a DNA strand, such that the DNA polymerase functions to excise mismatched nucleotides from the 3' terminus of the DNA strand at a faster rate than the rate at which the DNA polymerase functions to remove nucleotides matched correctly with the nucleotides of the template, and which DNA polymerase does not exhibit 5'-3' exonuclease activity. The invention also relates to isolated and cloned DNA sequences derived from the Bacillus stearothermophilus thermostable DNA polymerase, as well as the expressed polymerase itself.

Abstract: The present invention is directed to a process for amplifying and detecting any target nucleic acid sequence contained in a nucleic acid or mixture thereof and for assembling large polynucleotides from component polynucleotides, each involving generating concatemers formed by PCR amplification of overlapping fragments.

Abstract: Methods for selecting transition metal complexes (TMCs) that can modulate the translation of an mRNA by selectively cleaving a regulatory element of the mRNA are described. Included are in vitro, in situ, and/or in vivo methods. In addition, methods of identifying and characterizing mRNAs that contain regulatory elements which can be targeted by the TMCs of the present invention are also provided. Pharmaceutical compositions containing the TMCs of the present invention are also included.

Abstract: Stabilized enzyme compositions for use in nucleic acid amplification. Compositions are provided for the stabilization of one or more enzymes in a single stabilized formulation. Additional compositions incorporate a dried, stabilized enzyme mixture together with necessary cofactors and enzyme substrates in a single container for use upon rehydration. Also disclosed are methods for making and using stabilized enzyme compositions and kits for nucleic acid amplification incorporating the disclosed compositions.

Abstract: The invention is directed to alkaline glucose oxidases comprising novel peptide sequences obtained from a strain of Cladosporium, particulary Cladosporium oxysporum, strain CBS 163.94. Furthermore, the invention relates to methods for producing and using said glucose oxidases.

Abstract: A process is provided whereby the constitution of starch produced in a plant is altered without there being a substantial change in the total amount of starch which is produced. In the process a plant cell is transformed using a chimaeric gene comprising an antisense coding sequence from the waxy locus of a plant genome or an antisense similar coding sequence from a non-plant genome.

Abstract: The invention provides methods and compositons for convergent synthesis of branched polymers useful as molecular probes. The invention also includes several novel branched polymeric structures particularly useful for detecting target polynucleotides. Branched polymers of the invention comprise at least two branches: at least one branch is a target binding moiety capable of specifically binding to a target molecule of interest and one or more branches are signal generation moities capable of directly or indirectly generating a detectable signal. In accordance with the method of the invention branched polymers are assembled from components having phosphorothioate groups and/or haloacyl- or haloalkylamino groups. The phosphorothioate and haloacyl- or haloalkylamino groups react rapidly and efficiently when brought into contact to form thiophosphorylacyl- or thiophosphorylalkylamino bridges which complete the assembly of a branched polymer.

Abstract: DNA fragments from Porphyromonas gingivalis which express proteins that elicit anti-P. gingivalis immunologic responses are described. Microorganisms, genetically modified to express P. gingivalis antigens, are provided. Also disclosed are probes, vaccines, and monoclonal antibodies for the detection and prevention of periodontal disease.

Abstract: Isolated mpl ligand, isolated DNA encoding mpl ligand, and recombinant methods of preparing mpl ligand are disclosed. These mpl ligands are shown to influence the replication, differentiation or maturation of blood cells, especially megakaryocyte progenitor cells. Accordingly, these compounds are used for treatment of thrombocytopenia.

Abstract: Disclosed is a method of treating a heterogeneous population of cells to link together copies of two or more nucleic acid sequences from at least some of the cells, the arrangement being such that copies of the DNA sequences from an individual cell are preferentially linked in the vicinity of the nucleic acid from which the copies are derived. Also disclosed are recombinant proteins expressed by the method of the invention and kits for performing said method.

Abstract: This invention provides a method to normalize a directional cDNA library constructed in a vector that allows propagation in single-stranded circle form comprising: (a) propagating the directional cDNA library in single-stranded circles; (b) generating fragments complementary to the 3' noncoding sequence of the single-stranded circles in the library to produce partial duplexes; (c) purifying the partial duplexes; (d) melting and reassociating the purified partial duplexes to appropriate Cot; and (e) purifying the unassociated single-stranded circles, thereby generating a normalized cDNA library. This invention also provides normalized cDNA libraries generated by the above-described method and uses of the generated libraries.

Abstract: The present invention relates to the cloning of the gene of a thermophilic DNA ligase, from Thermus aquaticus strain HB8, and the use of this ligase for the detection of specific sequences of nucleotides in a variety of nucleic acid samples, and more particularly in those samples containing a DNA sequence characterized by a difference in the nucleic acid sequence from a standard sequence including single nucleic acid base pair changes, deletions, insertions or translocations.

Abstract: The invention relates to the construction and use of novel expression systems that make use of DNA sequences encoding a promoter or promoters along with cis-regulatory elements, such as those for proU, that permit osmotically inducible initiation of transcription in bacteria; the expression systems of the present invention can be used to efficiently hyperexpress heterologous gene products including proteins or polypeptides; methods for construction of vectors and strains enabling NaCl-induced hyperexpression of heterologous gene products in several organisms such as Escherichia coli, and methods for NaCl-induced hyperexpression of heterologous gene products, including proteins or polypeptides, using the said novel combinations in a variety of organisms such as Escherichia coli, and further purification of the gene product of interest in either laboratory or industrial scale production systems.

Abstract: This invention herein describes pharmaceutical compositions and methods for targeted delivery of functional genes into cells and tissues in vivo. The invention discloses DNA:lipid complexes, methods of making such complexes and methods of using such complexes for facilitating the targeted delivery and entry of recombinant expression constructs into cells and tissues in vivo, and particularly delivery of such recombinant expression constructs by intravenous, intraperitoneal or direct injection.

Abstract: Methods are provided for the production of large polypeptides containing repeating sequences of amino acids utilizing biochemical techniques, specifically DNA sequences coding for the expression of the large polypeptides. Systems utilizing exogenous transcriptional and translational regions to control the production of the large polypeptides are also provided.

Abstract: A method and apparatus for selectively applying a print material onto a substrate for the synthesis of an array of oligonucleotides at selected regions of a substrate. The print material includes a barrier material, a monomer sequence, a nucleoside, a deprotection agent, a carrier material, among other materials. The method and apparatus also relies upon standard DMT based chemistry, and a vapor phase deprotection agent such as solid TCA and the like.

Abstract: A novel method for inactivating undesirable members in a nucleic acid sample which comprises undesirable and desirable members of a nucleic acid family. The method includes the step of adding a blocker to the sample which selectively associates with the undesirable members such that they are unable to participate in any further processing of the sample. The method advantageously provides a means whereby desirable members of the nucleic acid family can be processed, i.e. identified, amplified and/or isolated, without interference from undesirable sequence-related members.

Abstract: Method for the sensitive detection of a target nucleic acid by hybridization with a probe nucleic acid. The latter contains a part which can hybridize with the target nucleic acid and a nucleic acid-specific part which does not hybridize with the target nucleic acid. The method further comprises cleavage of the probe nucleic acid, hybridization of a cleavage product of the probe nucleic acid containing the part that does not hybridize with the target nucleic acid with a matrix nucleic acid containing a part that can be hybridized with the cleavage product and a part that cannot be hybridized with the probe nucleic acid. The method also comprises the determination of the hybrid consisting of the cleavage product and the matrix nucleic acid and a reagent kit suitable for this purpose.

Abstract: Apparatus and method for performing a nucleic acid amplification reaction and preferably a polymerase chain reaction (PCR) in a reaction mixture in at least one capillary tube. Several different embodiments are disclosed. One embodiment cycles a sample through a capillary tube loop passing through two thermostatted fluid baths. Another embodiment has the capillary tube routed alternatingly between two heat exchangers to that the sample makes only one pass through the tube. Other embodiments maintain the heat exchangers stationary and translate the samples between them. Still further embodiments maintain the samples stationary and either automatically translate or rotate the heat exchangers past the samples contained within the capillary tubes to perform the thermal cycles necessary for the amplification reaction.