Abstract: A stable xenogeneic fusion partner that is a product of the fusion of a mouse myeloma cell and a non-transformed rabbit partner cell. The fusion partner produces undetectable levels of antibody, as determined by an enzyme linked immunosorbent assay. A method for generating rabbit monoclonal antibodies is disclosed that comprises fusing a nontransformed rabbit partner cell with a rodent myeloma cell to produce a xenogeneic fusion partner, selecting a stable fusion partner producing undetectable levels of antibodies, fusing the stable fusion partner with a rabbit antibody producing cell and isolating an antibody producing cell line that produces antibodies directed to a predetermined antigen.

Abstract: A hydrogen bond labeling and base sequence determination method for DNA or RNA takes advantage of the complementary nature of the fundamental structure of DNA or RNA bases to bind corresponding chemical species with the base species of a denatured DNA or RNA chain. After providing an aqueous solution containing a nucleic acid, heating the aqueous solution to cleave the nucleic acid; cooling the aqueous solution and adding a base-specific labeling molecule having available hydrogen bonds to the aqueous solution, the base-specific labeling molecules bond to the available bases in a one-to-one fashion via the available hydrogen bonds. The resulting labeled molecule can be microscopically observed to determine the base sequence of the nucleic acid.

Abstract: Methods for amplifying DNA sequences of interest are disclosed. The methods can be performed using only one primer and are also useful in cloning protocols and for sequencing large DNAs. The methods comprise cleaving a sample DNA using an agent, such as a restriction endonuclease, that produces discrete DNA fragments; ligating the fragments to "adapter" polynucleotides having a ligatable end and first and second self-complementary sequences separated by a spacer sequence, thereby forming ligated duplexes; denaturing the ligated duplexes to form templates; annealing molecules of an oligonucleotide primer to the templates, the primers being homologous to a primer target site associated with the sequence of interest; extending the primers using a DNA polymerizing agent to form duplex products; and denaturing the duplex products. Subsequent multiple cycles of annealing primers, extending the primers, and denaturing duplex products are usually performed so as to achieve the desired degree of amplification.

Abstract: A method is provided for detecting, ordering and mapping genes or DNA sequences in the genome of eukaryotic cells. The method comprises the steps of releasing free chromatin from the nuclei of the cells and contacting the released free chromatin with at least one detectable probe capable of hybridizing to the genes or DNA sequences to be detected, thereby rendering the genes or DNA sequences detectable.

Abstract: Appendicitis can be detected in human beings suspected of having appendicitis by determining a threshold level of 10 mg/liter of .sigma.-hydroxyhippuric acid in the urine of such humans. This threshold level can be determined by qualitative, semiquantitative, or quantitative methods including HPLC (high pressure liquid chromatography), TLC (thin layer chromatography), radioimmunoassay, colorimetric tests, NMR (nuclear magnetic resonance), mass spectrometry, electrophoresis, monoclonal antibody tests, and enzymatic tests. The absence of appendicitis can be established by the presence of .sigma.-hydroxyhippuric acid in concentrations less than about 10 mg/liter in a urine sample.

Abstract: The present invention is directed to a gene encoding an envelope glycoprotein of equine herpesvirus type 1 (EHV-1), the glycoprotein D (gD) gene, its gene product and antibodies directed against gD polypeptides. The envelope glycoproteins of herpesvirus are major targets of the immune response to herpesviral infection. Hence, an important aspect of this invention is directed towards a vaccine against EHV-1 and treatment of EHV-1 infection by anti-EHV-gD antibodies or antisera.

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: Method and composition for detecting one or more selected polynucleotide regions in a target polynucleotide. In the method, a mixture of sequence-specific probes are reacted with the target polynucleotide under hybridization conditions, and the hybridized probes are treated to selectively modify those probes which are bound to the target polynucleotide in a base-specific manner. The resulting labeled probes include a polymer chain which imparts to each different-sequence probe, a distinctive ratio of charge/translational frictional drag, and a detectable label. The labeled probes are fractionated by electrophoresis in a non-sieving matrix, and the presence of one or more selected sequences in the target polynucleotide are detected according to the observed electrophoretic migration rates of the labeled probes in a non-sieving medium.

Abstract: A method and kit are provided for detecting the interaction between a first test protein and a second test protein, in vivo, using reconstitution of the activity of a transcriptional activator. This reconstitution makes use of chimeric genes which express hybrid proteins. Two types of hybrid proteins are prepared. The first hybrid contains the DNA-binding domain of a transcriptional activator fused to the first test protein. The second hybrid protein contains a transcriptional activation domain fused to the second test protein. If the two test proteins are able to interact, they bring into close proximity the two domains of the transcriptional activator. This proximity is sufficient to cause transcription, which can be detected by the activity of a marker gene which contains a binding site for the DNA-binding domain.

Abstract: Oligonucleotides (SEQ ID NOs 1-8) selectively hybridizable with a specific gene of Vibro parahaemolyticus, oligonucleotides (SEQ ID NOs 9-13) selectively hybridizable with the LT gene of toxigenic Escherichia coil, oligonucleotides (SEQ ID NOs 14-21) selectively hybrizable with the STh or STp gene of toxigenic Escherichia coil, oligonucleotides (SEQ ID NOs 22-47) selectively hybridizable with the entA, B, C, or D gene of Staphylococcus aureus, or oligonucleotides (SEQ ID NOs 48-53) selectively hybridizable with the entE gene of Staplyloccus aureus are prepared and used as primers for gene amplification to thereby selectively detect only respective microorganisms causing food poisoning.

Abstract: Hematopoietic cell populations are separated to provide cell sets and subsets as viable cells with high purity and high yields, based on the number of original cells present in the mixture. High-speed flow cytometry is employed using light characteristics of the cells to separate the cells, where high flow speeds are used to reduce the sorting time.

Abstract: Nucleic acid probes are described for detecting yeasts capable of causing cryptococcosis, specifically Cryptococcus neoformans. The preferred probes are complementary to the ribosomal ribonucleic acid sequences unique to Cryptococcus neoformans, and as such can detect the rRNA, rDNA, or polymerase chain reaction amplification products of these genes. The detection of the etiological agent of human cryptococcosis, and tests for making a clinical diagnosis of this disease utilizing specific rRNA or rDNA probes are now possible.

Abstract: The invention is directed to a method of purifying sequentially synthesized peptides and oligonucleotides by affinity techniques. Selected products are capped with and N-terminus capping agent for peptides or a 5'-terminus capping agents for oligonucleotides, and then bound with affinity agents that are selective for the corresponding capping agents.

Abstract: Disclosed herein is a composition comprising ribonuclease treated RNA or DNA dependent RNA polymerase, and the use of same in amplification methods. The treatment of the RNA or DNA dependent RNA polymerase with ribonuclease reduces or eliminates false positives which result from the presence of an endogenous or contaminating replicatable template species in the Q.beta. replicase enzyme preparation.

Abstract: A method for determining the nucleic acid sequence that encodes ligand binding domain on any protein encoded by a known gene includes the steps of non-specifically cleaving the coding region of the known gene, size-selecting the cleavage products, cloning the size-selected gene products into an expression vector active in a heterologous host, using a novel cloning strategy expressing the peptides produced by the clones and screening for clones that produce peptides that interact with an antibody or ligand of interest.

Abstract: For the detection and quantitative determination of nitrosomonas strains in wastewaters and soils, a gene probe is used which, by virtue of its complementary sequences, only hybridizes with parts of the genome of nitrosomonas strains from wastewater or soil samples and does not produce a positive hybridization signal with parts of the genome of other bacteria and the hybridized nucleic acid is quantitatively determined by means of a known label of the gene probe and thus provides a direct measure of the content of nitrosomonas strains in the wastewater or soil sample.

Abstract: The present invention provides PCR primers with which sexing of bovine embryos can be easily attained and provides a practical, rapid and reliable method for determining the sex of bovine embryos using these primers. The methods for determining the sex of the bovine embryos are characterized by discriminating PCR products which are obtained by amplifying specific DNA sequences by PCR with pairs of male-specific and gender-neutral primers. These primers are derived from DNAs which specifically hybridize to the bovine male genome and from DNAs which gender-neutrally hybridize to both bovine male and female genomes.

Abstract: The present invention relates to a method for detecting base sequence differences within homologous regions of two DNA molecules comprising the steps of contacting at least one strand of the first DNA molecule with the complementary strand of the second DNA molecule under conditions such that base pairing occurs, contacting the resulting DNA duplexes with a protein that recognizes substantially all base pair mismatches under conditions such that the protein forms specific complexes with its cognate mispairs, and detecting the resulting DNA:protein complexes by a suitable analytical method. Also disclosed are protein components of DNA mismatch correction systems and the use of these components in methods for genetic mapping.

Abstract: Nucleic acid isolates capable of hybridizing only to the Y-chromosome specific DNA sequences of cattle, sheep and goats are described, as are methods for the determination of the sex chromosome constitution of a tissue or cell sample.

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.