Abstract: Methods for preparing liquid supports utilized in bioaffinity and ion-exchange separation methods and provided. The support is based on an inert carrier with ligands or binders for ligands attached through its surface through a highly fluorinated isocyanate anchor group. The use of such supports in capturing neutral and charged target molecules from samples and in analytical applications are also provided.

Abstract: A nucleic acid construct useful in preparing reagents for determining target nucleotide sequences in the nucleic acid of a biological sample, the construct having in its single-stranded form:(a) a target binding region substantially complementary to the target nucleotide sequence, and(b) a signal strand pairing segment bound in the construct by complementary base pairing to a portion of the target binding region;a second portion of the target binding region being single-stranded; andthe target binding region and signal strand pairing segment being covalently linked by a phosphate/sugar backbone.A replicable nucleic acid having an origin of replication and two half-restriction sites capable of forming a restriction site can be treated with a restriction enzyme to form a length of nucleic acid containing the target binding region and the signal strand pairing segment. Subsequent labeling of the construct and various optional cleavage and derivation steps can convert the construct to a reagent complex.

Abstract: Bioaffinity and ion-exchange separation methods are provided along with liquid supports utilized in these methods. The support is based on an inert carrier with ligands or binders attached to its surface through a highly fluorinated isocyanate anchor group. Methods for preparing such supports and their use in capturing neutral and charged target molecules from samples and in analytical applications are also provided.

Abstract: Unique species-specific Eimeria maxima DNA probes comprising divergent DNA sequences are disclosed. The probes are complementary to a small subunit ribosomal RNA gene of Eimeria maxima.

Abstract: Single-chain analogs of the naturally occurring two-chain peptide monellin retain the sweetening properties of the natural protein and are stable under conditions which would otherwise destabilize the native peptide. A covalent linkage joins peptides corresponding to portions of the A and B chains of the naturally occurring protein.

Abstract: The present invention relates to a novel cell line isolated from a rat osteosarcoma wherein the cell line has the following characteristics: a) a mutated p53 gene incapable of expressing p53 protein, b) a normal RB-1 gene, c) a 10-fold amplified c-myc gene, d) a normal c-fos gene, e) a deregulated immediate early gene response, f) a canalicular network MATRIGEL.TM. growth pattern, g) tumorigenic in congenitally athymic mice, h) no alkaline phosphatase activity, i) an ability to produce one or more growth factors selected from the group consisting of: 1) an osteoblastic differentiation growth factor, 2) a non-heparin binding mitogenic growth factor, 3) a first heparin binding mitogenic growth factor, 4) a second heparin binding mitogenic growth factor, and 5) a third heparin binding mitogenic growth factor, and j) an ability to be serially propagated greater than sixty population doublings.

Abstract: Methods for the isolation and purification of the phytotoxin cercosporin are disclosed as well as methods for identifying microorganisms capable of degrading cercosporin. Cercosporin can be purified from members of the fungal genus Cercospora and incorporated into culture medium for selection of those organisms resistant to cercosporin. Once identified, these organisms can be used to isolate the protein and the gene responsible for conferring cercosporin-resistance. The gene can be cloned and inserted into a suitable expression vector so that the protein can be further characterized. Additionally, the DNA encoding for cercosporin-resistance can be inserted into a vector suitable for transforming an Agrobacterium and the Agrobacterium in turn used to transform plant cells normally susceptible to Cercospora infection. Plants can be regenerated from the transformed plant cells. In this way, a transgenic plant can be produced with the capability of degrading cercosporin.

Abstract: A probe/primer which includes a substantially purified single-stranded oligonucleotide containing a region the sequence of which is identical to the sequence of a six-nucleotide, single-stranded segment of a gene encoding a mutant form of a human photoreceptor protein, which segment includes the mutation; and methods of making and using such probe/primer.

Abstract: Differences in the rate of kynurenine formation in the lysates of red blood cells of a patient suspected of having Tourette syndrome, Tourette syndrome associated disorders or Tourette spectrum disorders and of a control are used for the diagnosis of such disorders.

Abstract: The present invention relates to a gene encoding a new member of the metalloproteinase family which has been found to be specifically associated with invasive breast cancer, and to methods of diagnosis for such cancer comprising detection of the marker or its nucleotide sequence, and to treatment or prophylaxis by inhibiting, altering the activity of or binding the marker, or by interfering with its synthesis.

Abstract: A point mutation in the CD18 gene responsible for causing bovine leukocyte adhesion deficiency (BLAD) and a silent mutation linked thereto have been identified. Nucleic acid sequences encompassing these mutations serve as bases for testing and identifying cattle alleles attributable to BLAD using any of a variety of diagnostic assays. Suitable nucleic acid probes and primers have also been designed for use in such assays.

Abstract: Probes for the detection of Streptococcus pyogenes, which are capable of distinguishing it from related species, are provided. Methods of using these probes in hybridization assays, and hybrids formed between the probes and complementary nucleic acids, are disclosed.

Abstract: This invention provides a rapid and highly effective method for extracting nucleic acids from cells or virions without the use of proteolytic enzymes. Extraction is accomplished within a few minutes using a lysing composition comprising a buffer, a source of a DNA polymerase cofactor, a stabilizer and at least one nonionic surfactant which will release nucleic acids from cytoplasmic and nuclear membranes of cells or virions. The resulting mixture is heated to boiling for up to fifteen minutes, and the nucleic acids are recovered for amplification using polymerase chain reaction. No proteolytic enzyme is used in the extraction process.

Abstract: A nucleic acid comprising a base sequence which codes for a CEA family member 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 injected with such cloning vehicles, polypeptides expressed by such cells, synthetic peptides derived from the coding sequence of CEA family member nucleic acids, antibody preparations specific for such polypeptides, immunoassays for detecting CEA family members using such antibody preparations and nucleic acid hybridization methods for detecting CEA family member nucleic acid sequences using a nucleic acid probe comprising the above described nucleic acid.

Abstract: Genotoxic chemicals are an existing wide-spread health hazard to the human population. Advances in genetic toxicology testing have made it possible to assay potential mutagens, carcinogens, teratogens and clastogens in the environment. The mouse micronucleus assay provides an example of an excellent test for genetic damage to cells. When chromosome breaks occur in the blood stem cell population, the damaged piece of chromosome remains behind as a micronucleus in the normally DNA deficient red blood cells. However, currently available manual micronucleus assays are costly, time consuming, and labor intensive. In addition, the statistics are often marginal since the number of micronucleii (MNs) in 1000 polychromatic cells are scored manually, yielding limited amounts of data. This invention discloses the means for assaying the change in micronucleated cells by high speed flow cytometry.

Abstract: A probe/primer which includes a substantially purified single stranded oligonucleotide containing a region the sequence of which is identical to the sequence of a six-nucleotide, single-stranded segment of a gene encoding a mutant form of a human photoreceptor protein, which segment includes the mutation; and methods of making and using such probe/primer.

Abstract: The invention relates to a family of DNA insertion sequences (ISMY) of mycobacterial origin and other DNA probes which may be used a probes in assay methods for the identification of mycobacteria and the differentiation between closely related mycobacterial strains and species. In one method the probes are used to distinguish pathogenic M. paratuberculosis from M. avium, which finds an application in the diagnosis of Crohn's disease in humans and Johne's disease in animals. The use of ISMY, and of proteins and peptides encoded by ISMY, in vaccines, pharmaceutical preparations and diagnostic test kits is also disclosed.

Abstract: This invention concerns a coordination complex and salts and optically resolved enantiomers thereof, of the formula (R).sub.3 --M, wherein R comprises 1,10-phenanthroline or a substituted derivative thereof, M comprises a suitable transition metal, e.g. ruthenium(II), RHODIUM(III) or cobalt(III), and R is bonded to M by a coordination bond.The complexes of this invention are useful in methods for labeling, nicking and cleaving DNA. The lambda enantiomer of complexes of this invention is useful in methods for specifically labeling, detecting, nicking and cleaving Z-DNA or A-DNA.The complexes may also be used in a method for killing tumor cells and may be combined with a pharmaceutically acceptable carrier to form a pharmaceutical composition for the treatment of tumor cells in a subject. The invention further concerns methods for treating a subject affilicted with tumor cells.

Abstract: Hepatitis delta is used as a vector for inhibition of viral infection and to express proteins in vivo in a cell-specific manner. The scope of delta's use as a vector is broadened in the present invention in several important ways. For example, a delta RNA genome capable of self-replication is enlarged to carry additional information, either coding for messenger RNA for a protein, or for a targeted ribozyme, which can be delivered to liver cells using delta's normally infectious properties, or to other cell types using chimeric delta viral agents carrying altered surface proteins. In another embodiment, the delta vector is made self-limiting, so that its role in delivering targeted information is separated from its viral property of unlimited infectious replication. Targeting is achieved through the use of sequences flanking the delta sequences that have affinity for sites on RNA to be cleaved.

Abstract: Ribozymes, sequences cleaving RNA, derived from sequences present in the hepatitis delta virus, have been engineered for greater specificity without increasing size. The specific ribozyme sequences are useful as reagents for cleaving RNA for experimental studies as well as antiviral therapies. Examples demonstrating the targeting of these sequences against HIV and Crohn's disease are described in detail. The sequences are also useful as diagnostics for the detection of hepatitis delta virus in tissue and fluid samples, as in blood banking, as well as in isolation and characterization of new viroids having ribozyme activity, using an RNA-specific hybridization method. Based on analysis of the two domain structure of the hepatitis delta virus, it is possible to construct a vector for expression of non-hepatitis delta virus proteins in mammalian cells.