Abstract: A method of preparing biologically active human myeloperoxidase including the steps of preparing a vector for the expression of human myeloperoxidase. The vector includes plasmid pNIV2703. The plasmid includes a HindIII-SnaBI, HindIII-EcoRV or HindIII-HpaI cassette, the cassette including a complete DNA sequence for hMPO as shown in FIG. 1. CHO cells are transformed with the vector. The cells are cultured. The biologically active human myeloperoxidase is recovered.

Abstract: This invention relates to the cloning and expression of autogenes encoding RNA polymerases of T7 and T7-like bacteriophages, in which the RNA polymerase gene is transcribed from a promoter which is recognized by the encoded RNA polymerase. Cloning of T7 autogenes was achieved by reducing the activity of the RNA polymerase sufficiently to permit host cell growth. T7 RNA polymerase activity was controlled by combining two independent methods: lac-repression of the recombinant lac operator-T7 promoter in the autogene and inhibition of the polymerase by T7 lysozyme. Expression systems for producing the RNA polymerases of T7 and other T7-like bacteriophages, and expression systems for producing selected gene products are described, as well as other related materials and methods.

Abstract: This invention provides a method of inhibiting the transcription of a gene, which is activated by AP-1 or an AP-1 component, comprising binding AP-1 or the component with a nuclear receptor so as to prevent the binding of AP-1 to the gene. The nuclear receptor can be the retinoic acid receptor, glucocorticoid receptor, vitamin D3 receptor, thyroid receptor, or estrogen receptor. Also provided is a composition of matter comprising AP-1 or an AP-1 component bound to a nuclear receptor. These methods and compositions can be used to treat arthritis and cancer.

Abstract: A method is provided for synthesizing catechol from a biomass-derived carbon source capable of being used as a host cell having a common pathway of aromatic amino acid biosynthesis. The method comprises the steps of biocatalytically converting the carbon source to 3-dehydroshikimate in said host cell, biocatalytically converting the DHS to protocatechuate, and decarboxylating the protocatechuate to form catechol. Also provided is a heterologous E. coli transformant characterized by the expression of genes encoding transhetolase, DAHP synthase, and DHQ synthase, further characterized by the constitutive expression of structural genes encoding 3-dehydroshikimate dehydratase and protocatechuate decarboxylase.

Abstract: A DNA preparation is provided which comprises a nucleotide sequence coding for at least a portion of the precursor polypeptide of ricin. Also provided are recombinant DNA molecules containing such a nucleotide sequence, as well as microorganisms transformed with such recombinant DNA molecules.

Abstract: The present invention pertains to a process for efficiently and accurately constructing indirectly high-resolution maps of genomes and other entities. Specifically, the process comprises the steps of generating three sets of reagents, namely, (1) sample points, (2) covering regions, and (3) probes; performing parallel experiments to efficiently compare these reagents and acquire two data tables, one of (a) probes with covering regions, and another of (b) covering regions with sample points; combining these two independent tables by means of an inner product operation to produce a third table that indirectly compares probes with sample points; analyzing this computed table to construct a high-resolution map of the probes, the sample points, and the covering regions. The resulting integrated map is then used to efficiently search for probes based on their proximity to selected sample points. With genome maps, this search can enable the rapid discovery of genes, their products, and their useful applications.

Abstract: The present invention relates to a method for enhancing colonization of desired microorganisms in a localized environment comprising introducing into said localized environment a genetically engineered microorganism comprising a gene comprising a DNA sequence coding for one or more substrate utilizing protein, wherein said substrate utilizing protein confers a competitive effective ability to utilize a substrate by a microorganism in which it is genetically engineered, as well as the isolated genes and the genetically engineered microorganisms of such a method.

Abstract: This invention concerns recombinant RNA molecules comprising a recognition sequence for the binding of an RNA-directed RNA polymerase, a sequence for the initiation of product strand synthesis-and a heterologous sequence of interest inserted at a specific site in the internal region of the recombinant molecule. Such recombinant RNA molecules are capable of serving as a template for the synthesis of complementary single-stranded molecules by RNA-directed RNA polymerase. The product molecules so formed are also capable of serving as a template for the synthesis of additional copies of the original recombinant RNA molecule. In a preferred embodiment of the invention Q.beta. replicase is used as the RNA-directed RNA polymerase. The invention also concerns methods of selectively cleaving RNA molecules at specific sites, methods of constructing recombinant RNA molecules, and methods for the autocatalytic in vitro replication of recombinant RNA molecules.

Abstract: A vector capable of integrating into the chromosome of Salmonella including a first DNA sequence encoding a heterologous protein, a second DNA sequence encoding a marker, and a third DNA sequence encoding a phoP regulatory region regulated gene product necessary for virulence, the third DNA sequence being mutationally inactivated.

Abstract: A process for providing cDNAs containing full primary structure information of proteins by selectively synthesizing full-length cDNAs containing sequences starting from the capped region of mRNAs is disclosed. The invention provides a process for producing an intermediate for the synthesis of a full-length cDNA, which comprises the steps of treating mRNA extracted from cells to eliminate the phosphate group from the 5' end of an uncapped degraded mRNA; decapping from the 5' end of a capped intact mRNA; and ligating either a DNA oligonucleotide or a DNA-RNA chimeric oligonucleotide represented by the following general formula [I] to the 5' end phosphate group formed in the above step by the action of T4 RNA ligase, thereby selectively adding either the DNA oligonucleotide or the DNA-RNA chimeric oligonucleotide having an arbitrary sequence to the 5' end of the intact mRNA.5'-dN1-dN2- . . . -dNm-N1-N2- . . .

Abstract: The isolated DNA sequence for the VP2 antigenic region of IBDV, GLS strain, is expressed by incorporation in a virus carrier. Four variant DNA sequences encoding the VP2 region are provided, each of which may be used in the preparation of a vaccine to prevent IBDV in poultry.

Abstract: Novel Bacillus thuringiensis toxins with hymenopteran activity are described.

Abstract: The present invention describes a purified and isolated nucleic acid molecule which encodes for the biosynthetic pathway of tetracycline, chlortetracycline or an analogue thereof. The invention relates to the isolation and cloning of the nucleic acid molecule in an isolated fragment from Streptomyces aureofaciens and the expression of the biosynthetic gene in a heterologous host such as Streptomyces lividans.

Abstract: Processes are provided for removal of cells as cell pellets from liquid milk samples, or from cultures or extracts of other food materials or other materials of biological origin. The concentrated cells in the pellet can be analyzed by various techniques to determine the relative cell count, such as by lysing of the cells followed by measurement of ATP. Nucleic amplification, as by the polymerase chain reaction method, can be carried out using the cellular pellet directly, without need for isolation of nucleic acid from the cells. After the amplification, an assay can be carried out for amplified nucleic acid segment indicative of the presence of cells of interest in the sample. The invention thus provides methods for obtaining cellular components from samples of milk, and cultures or extracts of other materials, including food materials, and for determining relative contamination of milk and such other materials by microorganisms. The invention also provides kits for carrying out its various methods.

Abstract: An expression vector for expressing a protein or polypeptide in a bacterium, which comprises a first DNA sequence encoding at least a secretion signal of a lipoprotein, and a second DNA sequence encoding a protein or fragment thereof, or polypeptide or peptide heterologous to the bacterium which expresses the protein or fragment thereof, or polypeptide or peptide. The bacterium expresses a fusion protein a lipoprotein or lipoprotein segment and the protein or fragment thereof, or polypeptide or peptide heterologous to the bacterium which expresses the protein or fragment thereof, or polypeptide or peptide. Such expression vectors increase the immunogenicity of the protein or fragment thereof, or polypeptide or peptide by enabling the protein or fragment thereof, or polypeptide or peptide to be expressed on the surface of the bacterium. Bacteria which may be transformed with the expression vector include mycobacteria such as BCG.

Abstract: Mutant plasmids of Rts1 and P1 are provided which permit for high copy numbers of the plasmid in host bacteria. The mutant plasmids also possess other improved characteristics, including the ability to coexist with other vectors in host bacteria, such as E. coli, the ability to control copy numbers, within the range of about 20 to 120, by the use of different host bacteria growth medium, and the ability to remove the plasmids from host bacteria by a simple shift in the bacteria growth temperature. These plasmids may also contain an additional nucleotide sequence, GH.sub.3, which confers increased temperature stability.

Abstract: A method for the identification of compounds as inhibitors of oncoprotein action is described. A compound is identified as an inhibitor of oncoprotein action based on its ability to modulate the transcription of a reporter gene. Transcription of the reporter gene is dependent upon the binding of a fusion oncoprotein to a specific DNA binding element positioned upstream of the reporter gene promoter. The fusion oncoprotein contains a transcriptional regulatory domain from an oncoprotein and a DNA binding domain from a different protein.

Abstract: A covalently linked conjugate of an oligonucleotide (ODN) with a peptide and a carrier or targeting ligand (ODN-peptide-carrier) includes a therapeutic oligonucleotide which is capable of selectively binding to a target sequence of DNA, RNA or protein inside a target cell. The ODN is covalently linked to a peptide which is capable of being cleaved by proteolytic enzymes inside the target cell. The peptide, in turn is covalently linked to a carrier or targeting ligand moiety which facilitates delivery of the entire ODN-peptide-carrier conjugate into the cell, and preferably into a specific target tissue type. Inside the cell, the peptide is cleaved, releasing the ODN which, by binding to the target DNA, RNA or protein sequence, brings about a beneficial result.

Abstract: A DNA element which was isolated from the upstream regulatory region of the CT/CGRP gene and which exhibits cell-specific enhancement of transcription is described. The enhancer activity of the element is further regulated by members of the superfamily of steroids and retinoids which increase or decrease transcription in a cell-specific manner. Methods of gene regulation and therapeutic uses involving the DNA element are also described. Furthermore, proteins which bind to the element and regulate transcription of genes under the control of the DNA element are also described. These proteins can be used for purposes of regulation of gene expression.

Abstract: A method is provided for sizing DNA fragments using high speed detection systems, such as flow cytometry to determine unique characteristics of DNA pieces from a sample. In one characterization the DNA piece is fragmented at preselected sites to produce a plurality of DNA fragments. The DNA piece or the resulting DNA fragments are treated with a dye effective to stain stoichiometrically the DNA piece or the DNA fragments. The fluorescence from the dye in the stained fragments is then examined to generate an output functionally related to the number of nucleotides in each one of the DNA fragments. In one embodiment, the intensity of the fluorescence emissions from each fragment is linearly related to the fragment length. The distribution of DNA fragment sizes forms a characterization of the DNA piece for use in forensic and research applications.