Abstract: Cloning systems useful for the isolation of recombinant nucleic acid are disclosed in which the recombination of cloning-system nucleic acid and foreign nucleic acid is linked to the expression of a moiety on the surface of a host organism, the moiety being a first member of a binding pair. When recombination occurs between the nucleic acid and the foreign nucleic acid, the moiety is expressed on the surface of the host organism. The isolation of recombinant nucleic acid is then performed by attaching a second member of the binding pair to a solid support and contacting the host organism with the support. When the first member of the binding pair is expressed on the surface of the host organism, the host organism binds to the second member of the binding pair attached to the solid support, thereby selectively isolating those organisms.

Abstract: The invention relates the discovery that the polymorphisms in 16S rRNA genes may be used to identify unknown bacterial isolates at the level of species, subspecies and strain identification. The invention also relates to the discovery that the sequences of individual are required. The invention involves the generation of composite polynucleotide sequence of the 16S rRNA genes of a microorganism of interest. The invention is to provide methods for identifying microorganisms. The methods comprise the steps of generating a composite 16S rRNA region sequence from a microorganism of interest. The composite 16S rRNA region sequence reveals polymorphisms within the 16S ribosomal RNA gene of the microorganism. The composite 16S rRNA region sequence may then be compared with a list of previously obtained composite 16S rRNA region sequences from reference microorganisms so as to determine the species, subspecies, or strain of the microorganism of interest.

Abstract: The invention relates to novel chimeric phosphoramidate oligonucleotides and their use in primer-extension methods such as DNA sequencing and nucleic acid amplification. The subject chimeric phosphoramidate oligonucleotides have both N3'-phosphoramidate linkages and phosphodiester linkages. The invention includes methods of primer extension using the subject chimeric oligonucleotides as primers. Primer extension methods of interest include nucleic acid amplification reactions, e.g. PCR, and polynucleotide sequencing reactions. In the primer extension methods of the invention, a chimeric phosphoramidate oligonucleotide primer is annealed to a polynucleotide template. After annealing, the chimeric oligonucleotide primer is extended by joining a nucleotide to the 3' end of the primer by a DNA polymerase catalyzed reaction. Other embodiments of the invention include methods of primer extension using phosphoramidate linkage containing polynucleotide templates.

Abstract: A method is provided for determining aneuploidy of selected chromosomes. An important feature of the invention is the quantitative amplification of STR markers with repeat units of at least 3 nucleotides. The amplified STR DNA is separated by size and the respective quantities of the amplified components are determined and related to chromosome copy number. The highly polymorphic nature of the STR markers permits a more sensitive and reliable quantitative analysis of the amplified DNA.

Abstract: The invention relates to new methods of RNA isolation that exploit the surprising discovery that RNA may be differentially precipitated from DNA. The subject methods result in the formation of an RNA-containing precipitate that has an RNA content at least two fold enriched with respect to DNA, as compared with the RNA to DNA ratio of the solution from which the RNA-containing precipitate is derived. The invention also includes various methods of DNA isolation that employ the selective precipitation of RNA. The degree of RNA enrichment achieved is often much greater than two fold; enrichment by a factor of ten or greater is frequently obtained. By precipitating RNA from a solution, the RNA may be collected by simple procedures such as centrifugation or filtration, thereby avoiding the need to bind the RNA to solid phase. The collected RNA precipitate may then be solubilized.

Abstract: Reporter-quencher probe assays of nucleic acid amplification, such as PCR, are rendered more meaningful by the addition of internal control reagents. An internal control polynucleotide is amplified with internal control primers and the product is measured by correlation with increased fluorescence by polymerase mediated-exonuclease cleavage or hybridization of the internal control probe. Probes specific for target and internal control polynucleotides are labelled with spectrally resolvable reporters, allowing for concurrent detection and measurement of target and control amplification. A kit of all PCR reagents can be dispensed into reaction chambers in a high-throughput system for rapid and accurate nucleic acid amplification assay, with real-time or end-point measurements. Fluorescent signals correlated to target and internal control levels are spectrally resolvable and measured concurrently.

Abstract: Methods of genotyping canines by analysis of polymorphisms in the number of microsatellite DNA repeats are provided. The internal repeat sequence is amplified by the use of specific primers. The number of repeats, and therefore the distance between the primers, is highly variable in a population, thereby providing an allelic marker for the locus. The combined information from multiple loci provides a means of distinguishing individuals, even among inbred dog breeds, for parentage testing, forensic testing and analysis of individuals relatedness.

Abstract: Cloning systems useful for the isolation of recombinant nucleic acid are disclosed in which the recombination of cloning-system nucleic acid and foreign nucleic acid is linked to the expression of a moiety on the surface of a host organism, the moiety being a first member of a binding pair. When recombination occurs between the nucleic acid and the foreign nucleic acid, the moiety is expressed on the surface of the host organism. The isolation of recombinant nucleic acid is then performed by attaching a second member of the binding pair to a solid support and contacting the host organism with the support. When the first member of the binding pair is expressed on the surface of the host organism, the host organism binds to the second member of the binding pair attached to the solid support, thereby selectively isolating those organisms.

Abstract: Cloning systems useful for the isolation of recombinant nucleic acid are disclosed in which the recombination of cloning-system nucleic acid and foreign nucleic acid is linked to the expression of a moiety on the surface of a host organism, the moiety being a first member of a binding pair. When recombination occurs between the nucleic acid and the foreign nucleic acid, the moiety is expressed on the surface of the host organism. The isolation of recombinant nucleic acid is then performed by attaching a second member of the binding pair to a solid support and contacting the host organism with the support. When the first member of the binding pair is expressed on the surface of the host organism, the host organism binds to the second member of the binding pair attached to the solid support, thereby selectively isolating those organisms. Other aspects of the invention include devices for the isolation of individual cells that differentially express binding moieties on their surface.

Abstract: Avian nucleotide sequences for use as sex specific markers are provided. Probes or primers derived from the sequences find use in defining the sex of a bird, by producing hybridization patterns or amplification products that are sex-specific. For a given avian species the probes may hybridize to both Z and W chromosomes so as to differentiate between the two chromosomes on the basis of restriction fragment length polymorphisms. Alternatively, the probes may hybridize exclusively to one of the two sex chromosomes in some species. The sequences taught are also useful for the isolation of other nucleotide sequences that are sex specific markers.

Abstract: This invention provides restriction endonuclease, Srf I, capable of recognizing the eight nucleotide base palindromic sequence shown below on a double-stranded DNA molecule and cleaving the DNA chain at the asterisk-marked position resulting in blunt ends5' GCCC*GGGC 3'3' CGGG*CCCG 5'(wherein C and G respectively represent cytosine and guanine). The restriction endonuclease is produced by culturing Streptomyces rf in a culture medium and recovering it from the culture.

Abstract: Murine monoclonal antibodies, or fragments thereof, that bind selectively to human breast cancer cells, are IgGs or IgMs, and when conjugated to ricin A chain, exhibit a TCID 50% against at least one of MCF-7, CAMA-1, SKBR-3, or BT-20 cells of less than about 10 nM. Methods for diagnosing, monitoring, and treating human breast cancer with the antibodies or immunotoxins made therefrom are described.

Abstract: This invention is in the area of immunology, and specifically relates to immunopharmacology as applied to the development of immunosuppressive compositions and methods of use thereof for treating a wide variety of diseases arising from abnormal or undesirable normal immune responses. Compositions and methods of using the same that are particularly useful in treating autoimmune diseases are shown.