Abstract: Compositions and methods are provided for modulating the expression of protein kinase C. Oligonucleotides are provided which are targeted to nucleic acids encoding PKC. The oligonucleotides contain a methoxyethoxy (--O--CH.sub.2 CH.sub.2 OCH.sub.3) modification at the 2' position of at least one nucleotide. Methods of inhibiting PKC expression and methods of treating conditions associated with expression of PKC using oligonucleotides of the invention are disclosed.

Abstract: The present invention provides a lyophilized reagent for PCR which is prepared by adding a stabilizing and sedimenting agent to an aqueous reaction mixture and lyophilizing thereof. The lyophilized PCR reagent of the present invention leads to a simplification of multi-step PCR manipulation, an increase of heat stability of the reaction mixture, prevention of carry-over contamination, and improved credibility of experiments. The lyophilized PCR reagent can be applied as a kit for analysis of DNA sequence or for diagnosis of diseases, which guarantee the results of high credibility in a short period of time.

Abstract: The invention relates to a thermocycler apparatus for implementing chemical and/or biological reactions and a method for activating the thermocycler apparatus. The thermocycler apparatus in accordance with the invention comprises a basebody in which for accommodating one or more reaction vessels open at the top an accommodating portion is configured. For closing off the accommodating portion of the basebody a cover is provided. Spring elements are arranged on the thermocycler apparatus such that the cover and the reaction vessel(s) are urged together and the reaction vessels are closed off directly by the cover or by means of an interlayer.The invention is characterized by an electrochemical linear motor arranged such that the cover and the reaction vessel(s) are urged together by a pressure higher than that produced by the spring.

Abstract: The invention concerns novel compounds having defined structural formulae and methods of mutating a nucleic acid sequence, the method comprising replicating a template sequence in the presence of a nucleoside triphosphate analogue in accordance with the invention, so as to form non-identical copies of the template sequence comprising one or more nucleoside triphosphate analogue residues, and a kit for use in performing the method of the invention.

Abstract: The present invention is directed to a mechanism for marking biological samples (blood, semen, saliva, etc.) that are to be used for subsequent nucleic acid analysis. The method involves adding a nucleic acid (DNA) molecule of known sequence to the biological sample at the time of sample collection. The method further utilizes primers specific to the complementary strands of the added DNA, such that they will direct the synthesis of another DNA molecule of known length when used in a standard or multiplex polymerase chain reaction (PCR). This provides an unambiguous identifying label for the collected forensic or medical samples, including blood, semen, saliva, urine, tissue, and mixtures of bodily fluids. When used with the supplied primers or DNA probe(s), PCR or nucleic acid hybridization techniques will produce or recognize DNA fragments of predetermined size(s), preventing errant confusion of said samples with other forensic or medical samples that do not contain the aforementioned DNA additive.

Abstract: Screening assays for identifying ligase activity modulators are provided, in both solid phase and liquid phase formats. Solid phase formats detect ligase activity by ligating a labelled nucleic acid to a capture nucleic acid in the presence of the ligase modulator and detection of the labelled nucleic acid. Liquid phase assays detect ligation-dependent changes in interactive labels between nucleic acids such as proximity quenching of fluorescent labels. Compositions, apparatus and integrated systems for assays are also provided.

Abstract: The present invention relates to improved methods for introducing nucleic acid into cells by first complexing the nucleic acid with a selected polycationic oligomer which neutralizes the negative charge of the nucleic acid, and then contacting the cell with the complex facilitating uptake of the nucleic acid into the cells as a complex with the oligomer. The methods are preferably applied to introduction of nucleic acid into eukaryotic cells, and more preferably into human cells. The invention also relates to methods of introducing antisense and triplex-forming oligonucleotides into prostate cancer cells to inhibit expression of proteins associated with (or that promote) malignancy and to inhibit cell growth or proliferation. More particularly, the invention relates to a method for inhibiting the expression of the HER-2/NEU protein in prostate cancer cells and to a method for inhibiting prostate cancer cell growth or proliferation.

Abstract: Methods and compositions for screening for intracellular transdominant effector peptides and RNA molecules selected inside living cells from randomized pools are provided.

Abstract: This invention concerns compositions and methods for the treatment of CMV infections. Antisense oligonucleotides are provided which are effective antiviral agents. In preferred embodiments, the oligonucleotides contain at least one 2'-methoxyethoxy modification and may be chimeric oligonucleotides.

Abstract: Cyanine dyes are used as the donor fluorophore in energy transfer labels in which light energy is absorbed by a donor fluorophore and transferred to an acceptor fluorophore which responds to the transfer by emitting fluorescent light for detection. The cyanine dyes impart an unusually high sensitivity to the labels thereby improving their usefulness in a wide variety of biochemical procedures, particularly nucleic acid sequencing, nucleic acid fragment sizing, and related procedures.

Abstract: The present invention is directed to a method for providing oligonucleotides or oligonucleotide analogs having known subunit sequences in which the desired oligomers are released from selected storage sites in one, two, or three dimensions, on a substrate by locally denaturing double-stranded complexes at the storage sites containing the desired oligomers. The released oligomers are useful in schemes for determining solutions to mathematical problems, in methods wherein hybridizing oligomers are used to encrypt and transmit data, in diagnostic and screening assay methodologies, and as primers or building blocks for synthesizing larger polynucleotides.

Abstract: Antisense compounds, compositions and methods are provided for modulating the expression of CD44. The compositions comprise antisense compounds, particularly antisense oligonucleotides, targeted to nucleic acids encoding CD44. Methods of using these compounds for modulation of CD44 expression and for treatment of diseases associated with expression of CD44 are provided.

Abstract: The present invention relates to the use of an osmolyte for reducing or abolishing non-covalent interactions of biological molecules to inert surfaces. Furthermore, the present invention relates to kits that may be employed for uses in accordance with the present invention.

Abstract: Modified oligonucleotides which possess at least one substituted 7-deazapurine base form more stable hybridization complexes with nucleic acids than unsubstituted analogs. They are useful as inhibitors of gene expression, as probes for detecting nucleic acids, as aids in molecular biology and as pharmaceuticals or diagnostic agents. Processes for preparing them are provided.

Abstract: The present invention relates to an antisense nucleic acid compound which has a nucleotide sequence complementary to at least 8 contiguous nucleotides in the nucleotide sequence of a gene coding for a vascular endothelial growth factor and which inhibits the expression of the vascular endothelial growth factor, as well as to a therapeutic or diagnostic agent for cancers, rheumatoid arthritis, diabetes etc., comprising said antisense nucleic acid as active ingredient. Further, the present invention relates to a method of preventing the expression of the vascular endothelial growth factor, comprising use of an antisense nucleic acid compound which has a nucleotide sequence complementary to at least 8 contiguous nucleotides in the nucleotide sequence of a gene coding for a vascular endothelial growth factor and which inhibits the expression of the vascular endothelial growth factor.

Abstract: A method of analyzing a polynucleotide target involves incubating the target with an oligonucleotide probe, generally an array of immobilised oligonucleotide probes, to form a duplex, and using ligase or polymerase to extend one chain of the duplex. A point mutation or variable number tandem repeat section may be analysed. Arrays of immobilised oligonucleotides are provided for use in the method.

Abstract: The present inventors have discovered that humans have a gene that encodes a novel protein of the thymosin .beta. family. This novel protein, herein referred to as thymosin .beta.15, has the ability to bind and sequester G-actin, like other members of the thymosin .beta. family, but unlike what is known about other members also directly regulates cell motility in prostatic carcinoma cells. A cDNA of the human thymosin .beta.15 gene (SEQ ID NO: 1) and having the deduced the amino acid sequence (SEQ ID NO: 2) was isolated. The present inventors have shown that enhanced transcripts (mRNA) and expression of the thymosin .beta.15 gene in non-testicular cells has a high correlation to disease state in a number of cancers, such as prostate, lung, melanoma and breast cancer, particularly metastatic cancers. Accordingly, discovering enhanced levels of transcript or gene product in non-testicular tissues can be used in not only a diagnostic manner, but a prognostic manner for particular cancers.

Abstract: Nucleic acid transporter systems for delivery of nucleic acid to a cell. The nucleic acid transporter includes a binding complex. The binding complex contains a binding molecule which non-covalently binds to the nucleic acid and covalently links to a surface ligand, nuclear ligand and/or a lysis agent. These may be linked to the binding molecule by spacers.

Abstract: A general method for screening genomic or cDNA, or fragments and mixtures thereof, involves sample simplification by the generation of subsets and then subjecting the subsets to a modified mismatch scanning procedure that eliminates DNA having single stranded breaks after a MutSLH cleavage. The methods are particularly useful in human population isolates, including identification of identical-by-descent sequences, genomic comparisons of two or more individuals, and genomic comparisons of two populations of individuals, for the identification of sequences of low polymorphism.

Abstract: The present invention makes available methods and reagents for novel manipulation of nucleic acids. As described herein, the present invention makes use of the ability of intronic sequences, such as derived from group I, group II, or nuclear pre-mRNA introns, to mediate specific cleavage and ligation of discontinuous nucleic acid molecules. For example, novel genes and gene products can be generated by admixing nucleic acid constructs which comprise exon nucleic acid sequences flanked by intron sequences that can direct trans-splicing of the exon sequences to each other. The flanking intronic sequences can, by intermolecular complementation, form a reactive complex which promotes the transesterification reactions necessary to cause the ligation of discontinuous nucleic acid sequences to one another, and thereby generate a recombinant gene comprising the ligated exons.

Abstract: Processes, kits and preferred devices for rapidly isolating large numbers of plasmid DNAs from plasmid containing cells and for performing high throughput DNA sequencing are described.

Abstract: A process is provided for the production of glycogens or an extract rich in glycogens from yeast cells, and a cosmetic composition containing them. A given quantity of yeast cells, from a specific culture or recovered as residues of a fermentation process, is subjected to an operation of enrichment in intracellular glycogens by culturing in two phases in the presence of a carbon source. The metabolism of the yeast cells is then stopped. The membranes of the yeast cells are then at least partially disintegrated to free intracellular substances, and the freed intracellular substances are subjected to at least one precipation to precipitate glycogens. A cosmetic composition comprising the glycogens is formulated in admixture with a dermatologically acceptable excipient.

Abstract: We sequenced a 625 and 617 bp fragment of the inner spacer region of 16S-23S rDNA of a strain of Acidovorax avenae representing pathogens from several hosts, including foxtail, oats, corn, rice, millet, sugarcane, orchid, and watermelon and a strain of A. avenae subsp. citrulli pathogenic only to watermelon, respectively, for the purpose of designing PCR primers for their identification. These plant pathogens were previously considered as non-fluorescent pseudomonads and have been recently reclassified as Acidovorax avenae subsp. avenae, A. avenae subsp. cattleyae, and A. avenae subsp. citrulli. Several sets of primers were designed. Primers identified by SEQ ID NO:1 and SEQ ID NO:2 of subsp. avenae reacted with all strains of A. avenae subsp. avenae (previously named P. avenae or P. alboprecipitans) originating from foxtail, oats, corn, rice, sugarcane, and millet, A. avenae subsp. cattleyae from orchid, and A. avenae subsp. citrulli (previously named P. pseudoalcaligenes subsp. citrulli) from watermelon.

Abstract: Reagents for the immobilization of biopolymers, processes for their preparation and their subsequent use in the immobilization of biopolymers for analytical and diagnostic procedures are described. One type of reagent includes a solid support fabricated of a polymeric material having at least one surface with pendant acyl fluoride functionalities. Another reagent includes solid supports fabricated of polymeric materials including ethylene acrylic acid or ethylene methacrylic acid copolymers and activated polypropylene. Processes for preparing reagents include derivatizing polymeric materials to form acyl fluoride functionalities or derivatizing ethylene acrylic acid copolymers and ethylene methacrylic acid copolymers to form active acyl functionalities.

Abstract: 2'-Deoxyisoguanosine, isosteric analogues and isoguanosine derivatives of formulae I-V, processes for their production via compounds of the general formulae a or b and reaction with aroyl isocyanates or from compounds of the general formulae VI-IX by photochemical irradiation. A further production process is the conversion of deoxyguanosines or guanosines by means of persilylation, reaction with ammonia and deamination in the 2 position. The compounds are suitable as pharmaceutical agents with antiviral efficacy.

Abstract: SAF-2 polypeptides and polynucleotides and methods for producing such polypeptides by recombinant techniques are disclosed. Also disclosed are methods for utilizing SAF-2 polypeptides and polynucleotides in therapy, and diagnostic assays for such.

Abstract: The present invention is directed to a method for preserving mycorrhizal fungi for long-term storage using lyophilization. This enables use of the fungi for growing and acclimatizing micropropagated plants. The invention is especially useful for preserving mycorrhizal ericoid fungi for long-term storage and use in a soilless medium for growing micropropagated ericaceous plants.

Abstract: The invention relates to the fabrication and use of biosensors comprising a plurality of optical fibers each fiber having attached to its "sensor end" biological "binding partners" (molecules that specifically bind other molecules to form a binding complex such as antibody-antigen, lectin-carbohydrate, nucleic acid-nucleic acid, biotin-avidin, etc.). The biosensor preferably bears two or more different species of biological binding partner. The sensor is fabricated by providing a plurality of groups of optical fibers. Each group is treated as a batch to attach a different species of biological binding partner to the sensor ends of the fibers comprising that bundle. Each fiber, or group of fibers within a bundle, may be uniquely identified so that the fibers, or group of fibers, when later combined in an array of different fibers, can be discretely addressed. Fibers or groups of fibers are then selected and discretely separated from different bundles.

Abstract: Methods of the invention comprise assays for markers indicative of cancer or precancer. Assays of the invention are performed on samples obtained from a patient by non-invasive or minimally-invasive methods. The invention provides nucleic acid indicia of cancer or precancer with high sensitivities and high specificities for detection.

Abstract: Double-stranded DNA molecules characterised in that they are circular and in that they essentially include one or more genes of interest.

Abstract: Chain reaction cloning methods and reagents and kits for performing such methods are provided. Chain reaction cloning allows ligation of double-stranded DNA molecules by DNA ligases and bridging oligonucleotides. Double-stranded nucleic acid molecules are denatured into single-stranded molecules. The ends of the molecules are brought together by hybridization to a template. The template ensures that the two single-stranded nucleic acid molecules are aligned correctly. DNA ligase joins the two nucleic acid molecules into a single, larger, composite nucleic acid molecule. The nucleic acid molecules are subsequently denatured so that the composite molecule formed by the ligated nucleic acid molecules and the template cease to hybridize to each. Each composite molecule then serves as a template for orienting unligated, single-stranded nucleic acid molecules. After several cycles, composite nucleic acid molecules are generated from smaller nucleic acid molecules.

Abstract: Novel HKID-1 polypeptides, proteins, and nucleic acid molecules are disclosed. In addition to isolated, full-length HKID-1 proteins, the invention further provides isolated HKID-1 fusion proteins, antigenic peptides and anti-HKID-1 antibodies. The invention also provides HKID-1 nucleic acid molecules, recombinant expression vectors containing a nucleic acid molecule of the invention, host cells into which the expression vectors have been introduced and non-human transgenic animals in which an HKID-1 gene has been introduced or disrupted. Diagnostic, screening and therapeutic methods utilizing compositions of the invention are also provided.

Abstract: A technique is described for identifying mutations, if any, present in a biological sample, from a pre-selected set of known mutations. The method can be applied to DNA, RNA and peptide nucleic acid (PNA) microarrays. The method analyzes a dot spectrogram representative of quantized hybridization activity of oligonucleotides in the sample to identify the mutations. In accordance with the method, a resonance pattern is generated which is representative of nonlinear resonances between a stimulus pattern associated with the set of known mutations and the dot spectrogram. The resonance pattern is interpreted to a yield a set of confirmed mutations by comparing resonances found therein with predetermined resonances expected for the selected set of mutations.

Abstract: Disclose is a method for making full-length cDNA libraries, which is for making libraries of cDNAs having lengths corresponding to full lengths of mRNAs and comprises the following steps of; forming RNA-DNA hybrids by reverse transcription starting from primers using mRNAs as templates, chemically binding a tag molecule to a diol structure present in the 5' Cap (.sup.7Me G.sub.ppp N) site of a mRNA which is forming a RNA-DNA hybrid, and separating RNA-DNA hybrids carrying a DNA corresponding to a full-length mRNA from the RNA-DNA hybrids formed above by using a function of the tag molecule. The present method is a method for preparing full-length cDNA libraries utilizing a method for labeling the 5' Cap site more efficiently than protein enzyme reactions, which is avoidable a decrease of a full-length cDNA synthesis efficiency caused by cleavage of mRNA, and can synthesize a full-length cDNA more efficiently.

Abstract: A general stochastic method for synthesizing random oligomers on particles is disclosed. A further aspect of the invention relates to the use of identification tags on the particles to facilitate identification of the sequence of the monomers in the oligomer.

Abstract: The present invention provides an improved method for the synthesis of double stranded DNA, particularly complementary DNA for the construction of directional complementary DNA libraries. The method comprises synthesizing a first strand of DNA complementary to a selected RNA or DNA template by contacting with the template a linker/primer comprising a selected restriction site, a suitable RNA or DNA dependent DNA polymerase, and substrates comprising a deoxyribonucleotide triphosphate analog. The linker/primer and deoxyribonucleotide triphosphate analog are selected such that incorporation of the nucleotide analog in the first strand substantially protects the double stranded DNA from cleavage, under conditions sufficient to cleave or substantially cleave the linker/primer, at the selected restriction site.

Abstract: Novel screening methods for identifying antimicrobial agents involving use of membrane potential indicator dyes are provided.

Abstract: Nucleic acid probes for detecting different strains of Citrus Tristeza Virus (CTV) were made and shown to be highly sensitive, specific, and selective. The invention also concerns a method of detection, a method of identifying novel strains of CTV, and a detection kit which employs the subject probes.

Abstract: The invention provides method and compositions for yeast cultures with a predetermined desirably improved trait, such an enhanced or reduced ability to produce particular fermentation products. In one embodiment, the breeding scheme involves assembling and surveying a large population of natural wine yeast for a chosen trait; select appropriate individuals from this population and repeatedly back-crossing to well-established commercial strain by selecting HO/HO spores from the selected and commercial parental strains, treating asci with zymolase and carrying out spore-spore pairings, observing the spore pairs at intervals to detect formation of primary zygotes, and selecting and sporulating primary zygote clones; analyzing segregants of primary zygote clones and scoring for the trait; selecting the best segregants; and repeating the back-crossing procedure using a different HO/HO spore clone from the commercial parent in each cross.

Abstract: The present invention is directed generally to methods facilitating the cloning of nucleic acid molecules. In particular, the invention relates to the use of polymerase inhibitors, including but not limited to anti-polymerase antibodies (such as anti-Taq antibodies) and fragments thereof, to inactivate residual polymerase activity remaining after the amplification (particularly via PCR) of a target nucleic acid molecule. The invention further provides compositions, particularly storage-stable compositions, comprising one or more components, such as one or more restriction endonucleases and one or more polymerase inhibitors, that are useful in cloning amplified or synthesized nucleic acid molecules by the above-described methods. The invention also relates to nucleic acid molecules produced by these methods, and to genetic constructs (such as vectors) and host cells comprising these nucleic acid molecules.

Abstract: Compositions and methods for the treatment and diagnosis of diseases or conditions amenable to treatment through modulation of expression of a gene encoding a p38 mitogen-activated protein kinase (p38 MAPK) are provided. Methods for the treatment and diagnosis of diseases or conditions associated with aberrant expression of one or more p38 MAPKs are also provided.

Abstract: DNA clones encoding a receptor in the Ig superfamily and a related soluble variant have been isolated from a human monocyte library. The invention provides receptor polypeptides, nucleic acids encoding them, expression vectors, and transformed cells for recombinant production of the polypeptides.

Abstract: Antisense compounds, compositions and methods are provided for modulating the expression of bc1-6. The compositions comprise antisense compounds, particularly antisense oligonucleotides, targeted to nucleic acids encoding bc1-6. Methods of using these compounds for modulation of bc1-6 expression and for treatment of diseases associated with expression of bc1-6 are provided.

Abstract: A process of amplifying a nucleic acid sequence by a procedure involving a polymerase chain reaction or a ligase chain reaction. The process involves repeated cycles of steps including a nucleic acid denaturing step and a nucleic acid synthesis step, the synthesis step being carried out under the action of an enzyme (a nucleic acid polymerase or ligase). The denaturing step and the synthesis step are carried out in different denaturing and synthesis reaction zones, respectively, and, during the repeated cycles, the enzyme is maintained in isolation from the denaturing reaction zone, and conditions or reagents required for the denaturing step are maintained in isolation from the synthesis reaction zone to the extent that the reagents and conditions required for denaturing do not impede the synthesis reaction to a substantial extent. The use of separate zones for the steps of the reactions means that an enzyme that is destroyed or degraded by the reagents and conditions required for denaturing (e.g.

Abstract: A method for detecting a target nucleic acid molecule is provided, comprising the steps of (a) reacting a mixture comprising (i) a target nucleic acid molecule; (ii) a single-stranded nucleic acid probe containing a scissile linkage; (iii) an enzyme capable of cleaving the probe portion of a double-stranded target-probe complex at the scissile linkage; and (iv) ribosomal protein and/or spermine, under conditions and for a time sufficient to allow the target nucleic acid and probe to hybridize to each other and form a double-stranded target-probe complex, followed by cleavage of the probe and cycling of the target to a new uncleaved probe, such that one or more portions of the cleaved nucleic acid probe are released from the target-probe complex; and (b) determining whether cleaved portions of the nucleic acid probe are produced, and thereby detecting the presence of the target nucleic acid.

Abstract: Antisense compounds, compositions and methods are provided for modulating the expression of methionine aminopeptidase 2. The compositions comprise antisense compounds, particularly antisense oligonucleotides, targeted to nucleic acids encoding methionine aminopeptidase 2. Methods of using these compounds for modulation of methionine aminopeptidase 2 expression and for treatment of diseases associated with expression of methionine aminopeptidase 2 are provided.

Abstract: A single molecule of single-stranded sample DNA (7) having a bead (5) at one end and a magnetic bead (6) at the other end is extended and fixed in the field of view of a fluorescent microscope by using a magnetic force (11) and a laser trap (3), and a primer (8) is bonded thereto, followed by elongation reaction (10) using polymerase. Only a single chemically modified nucleotide (9) labeled with at least one fluorophore which varies depending on the kind of the base is incorporated. Only the single fluorophore incorporated is measured as a fluorescence-microscopic image by evanescent irradiation (13) with exciting laser beams, and the kind of the base is determined from the kind of the fluorophore. The fluorophore labeling the nucleotide incorporated is released by evanescent irradiation (13) with ultraviolet laser beams (2), and the next nucleotide is incorporated. DNA sequencing is carried out by repeating the above procedure.

Abstract: Disclosed are methods, DNA sequences, vectors and cell lines useful for the rapid generation of stable mammalian cell lines expressing high levels of recombinant proteins.

Abstract: Disclosed is a method for synthesizing polynucleotide molecules such as genes or gene segments. A primer having 5' and 3' ends is incubated with a relatively shorter template having a 5' region non-complementary to the primer, a 3' region complementary to the 3' end of the primer, and a non-reactive 3' terminus to allow the 3' region of the template to anneal to the primer. The annealed product is reacted with at least one nucleotide in the presence of a template-dependent polynucleotide polymerase to produce a primer extended at its 3' end by at least one nucleotide complementary to the 5' region of the template. The extended primer is then dissociated from the template. The extended primer is further extended by repeating this cycle for sufficient cycles, wherein the templates and enzymes may differ from cycle to cycle, to obtain the object polynucleotide. Also disclosed are template libraries and kits containing said libraries for use in conjunction with the polynucleotide synthesis method.

Abstract: The invention describes a method of quantitatively detecting specific nucleotide sequences. Said method is essentially characterized in that a single-stranded nucleic acid, particularly mRNA, which has been isolated from a mixture, e.g. a biological sample, is hybridized in solution, with a polynucleotide sequence which is essentially complementary to the sequence to be determined; the nucleic acid is then immobilized on a solid phase, and the amount of bound hybrid is determined. It has proven to be particularly advantageous if the binding to the coated solid phase is accomplished with the aid of the specifically bindable chemical group which is coupled to the sequence to be determined or to the polynucleotide probe sequence via a linker.