Abstract: Oligonucleotides are provided which are targeted to nucleic acids encoding human c-raf and capable of inhibiting raf expression. The oligonucleotides may have chemical modifications at one or more positions and may be chimeric oligonucleotides. Methods of inhibiting the expression of human raf using oligonucleotides of the invention are also provided. The present invention further comprises methods of inhibiting hyperproliferation of cells and methods of treating abnormal proliferative conditions which employ oligonucleotides of the invention.

Abstract: Recombinant mycobacterial vaccine vehicles capable of expressing DNA of interest which encodes at least one protein antigen for at least one pathogen against which an immune response is desired and which can be incorporated into the mycobacteria or stably integrated into the mycobacterial genome. The vaccine vehicles are useful for administration to mammalian hosts for purposes of immunization. A recombinant vector which replicates in E. coli but not in mycobacteria is also disclosed. The recombinant vector includes 1) a mycobacterial gene or portions thereof, necessary for recombination with homologous sequences in the genome of mycobacteria transformed with the recombinant plasmid; 2) all or a portion of a gene which encodes a polypeptide or protein whose expression is desired in mycobacteria transformed with the recombinant plasmid; 3) DNA sequences necessary for replication and selection in E. coli; and 4) DNA sequences necessary for selection in mycobacteria (e.g., drug resistance).

Abstract: The invention provides a recombinant plasmid containing a sequence encoding any genes inserted between 5' and 3' self-cleavage ribozymes. The recombinant plasmid can be amplified in vivo as well as in vitro while growing the host cell. When obtaining RNA transcripts of the inserted sequence, the recombinant plasmid does not require a restriction enzyme digestion step (run-off transcription) since cis-acting ribozymes perform self-catalyzed cleavage at 5' and 3' sides of the inserted sequence once it is transcribed. In this specific example, the trans-acting RNA enzyme sequence is inserted between 5' and 3' cleavage ribozymes. However, the trans-acting ribozyme sequence in the recombinant plasmid can be replaceable with any other sequence (e.g., antisense RNA, RNAs of HIV-1, HDV and other RNA viruses etc.). This construct is especially useful since each unit, consisting of 5' processing ribozyme, inserted sequence, and 3' processing ribozyme, can be connected in tandem.

Abstract: Compounds having highly specific endoribonuclease activity are described. The compounds of this invention, also known as ribozymes, comprise ribonucleotides having two hybridizing regions with predetermined sequences capable of hybridizing with a target RNA, a region of defined sequence and a base paired stem region.

Abstract: Genes (and portions thereof) which are stress-inducible, e.g., by cold-shock which encode useful proteins. The proteins contribute to confer thermo-tolerance and/or contribute to confer low temperature tolerance to organisms, like eucaryotes or procaryotes. Typical genes encoding such proteins and homologous genes encoding proteins with equivalent properties are discussed. Nucleotide sequences encoding such proteins, recombinant replicable expression vehicles which comprise DNA constructs which encode such proteins and competent transformed organisms like eucaryotes are discussed. The production of valuable fermentation products and of biologically active proteins under conditions outside the normal or optimum physiological growth conditions are described.

Abstract: Discoveries are disclosed that show that certain mutations in different parts of the mechanism for regulation of independently replicating element replication can be combined in one expression independently replicating element to produce a runaway-replication phenotype that is suppressible by a diffusible factor from another independently replicating element co-resident in the host cell. According to the present invention, an expression independently replicating element combining known inducible promoters with this runaway-replication phenotype is used in combination with a independently replicating element that suppresses this runaway phenotype to establish a gene expression system that provides both controllable gene amplification and controllable induction of gene expression without the use of chemical inducers or temperature shifts. This expression system produces high yields of proteins in readily isolatable forms.

Abstract: A DNA fragment is disclosed which has connected to the downstream side of a first DNA sequence containing a region coding for the first ribozyme RNA a second DNA sequence containing a region coding for second ribozyme RNA capable of cleaving by self-processing the 3'-terminus site of the first ribozyme RNA subjected to transcription. A ribozyme having the 3'-terminus site thereof self-processed is obtained by effecting transcription of a RNA using as a template a recombinant vector obtained by recombination by the use of the DNA fragment.

Abstract: The present invention relates to the .beta.-glucuronidase (GUS) gene fusion system, and to the cloning and characterization of the .beta.-glucuronidase and glucuronide permease genes of Escherichia coli. It is based on the surprising discovery that gene fusions comprising the .beta.-glucuronidase gene may be effectively expressed in a wide variety of organisms to produce active .beta.-glucuronidase enzyme. Because of the abundance and availability of useful substrates for .beta.-glucuronidase enzyme, GUS gene fusions may serve as a superior reporter gene system as well as an effective means of altering cellular phenotype. In conjunction with recombinant glucuronide permease, which may be used to render host cells permeable to .beta.-glucuronidase substrates, the GUS gene fusion system offers almost unlimited applications in the fields of plant and animal genetic engineering.

Abstract: The invention relates to DNA sequences that code for the amino acid sequence of the thrombin inhibitor hirudin, hybrid vectors containing such DNA sequences, host cells transformed with such hybrid vectors, novel polypeptides with thrombin-inhibiting activity produced from such transformed host cells, processes for the manufacture of these DNA sequences, hybrid vectors and transformed host cells, and processes for the manufacture of these thrombin-inhibitors using the transformed host cells. The hirudin compounds that can be produced according to the invention have valuable pharmacological properties.

Abstract: L-tryptophan is produced by constructing a recombinant DNA composed of a vector DNA and DNA fragments bearing all of genetic information relating to the synthesis of DS, AS, PRT, PRAI, InGPS, TS and PGDH, introducing the recombinant DNA into a microorganism belonging to the genus Corynebacterium or Brevibacterium, culturing the microorganism in a medium, and recovering L-tryptophan accumulated in the culture.

Abstract: Methods and means for the construction of strains of yeast capable of producing cellulolytic enzymes, achieved by the transfer of chromosomal genes or cDNA copies of mRNAs coding for cellulolytic enzymes isolated from the fungus Trichoderma reesei to yeast cells using recombinant DNA vectors capable of replicating in yeast. The correct expression of these cellulolytic genes in yeast leads to the production of cellulolytic enzymes which are secreted from the cell. This allows the yeast to hydrolyze 3-1,4-glucan substrates such as cellulose.

Abstract: The invention relates to recombinant DNA molecules and to methods for producing proteins by means of said molecules. Particularly, the present invention relates to recombinant DNA molecules which are capable of being synthesized in Bacillus strain bacteria comprising the regulation and deleted non-functional signal sequence of the a-amylase gene of B. amyloliquefaciens, or a substantial part thereof, to which sequence a structural gene of any desired homologous or heterologous protein or peptide may be joined. These recombinant DNA molecules can be used, for example, to achieve intracellular expression of any desired protein or peptide in Bacillus strain bacteria.

Abstract: A plasmid which contains a mammalian cell-derived autonomously replicating sequence DNA, a promoter and a gene for peptide production inclusive of the translation initiation codon.

Abstract: The claimed invention provides a eukaryotic plasmid vector which upon expression provides the enzymes required for cysteine biosynthesis, the vector preferably allowing for expression of these enzymes and concomitant production of cysteine in ruminal mucosa cells. The vector of the instant invention is engineered using recombinant techniques to insert microbial genes encoding serine acetyltransferase (SAT) and O-acetylserine sulphydrolase (OASS), preferably the isolated Salmonella typhimurium genes cysE encoding for SAT and cysK or cysM encoding for OASS, driven by a promoter efficient in a eukaryotic host cell, such as the SV40 late promoter, into a eukaryotic cloning vector, such as Promega's pGEM-2. The construction of the instant vector also allows for an additional gene which upon expression produces human growth hormone.

Abstract: A process for cloning the chitinase gene of Vibrio parahemolyticus is provided, comprising the steps of cleaving the Vibrio parahemolyticus DNA with Sau3A, PSTI or other restriction enzyme, mixing the cleaved DNA fragments in the presence of pUC18 and T4 ligase to produce a composite plasmid, and inserting the composite plasmid in a DH5a strain of E. Coli.

Abstract: The present disclosure provides the complete primary amino acid, and underlying DNA, sequence for the 47-kilodalton surface immunogen of Treponema pallidum, subsp. pallidum. The sequence was obtained by using a combined strategy of DNA sequencing of the cloned gene as well as confirmatory N-terminal amino acid sequencing of the native antigen. An open reading frame corresponding to the 47-kDa antigen was comprised of 367 amino acid codohs, which gave rise to a calculated molecular weight for the corresponding antigen of about 40,701. Also disclosed are methods for preparing variant and mutant molecules having biologically similar attributes, as well as methods for preparing particular antigenic/immunogenic subportions of the 47-kDa protein. In particular aspects, antigenic/immunogenic subportions are identified by hydrophilicity analysis of the protein sequence.

Abstract: The present invention comprises novel recombinant DNA compounds which encode the .about.40,000 dalton adenocarcinoma antigen recognized by monoclonal antibody KS 1/4. Eukaryotic and prokaryotic expression vectors have been constructed that comprise novel KSA-encoding DNA and drive expression of KSA when transformed into an appropriate host cell. The novel expression vectors can be used to produce KSA derivatives, such as non-glycosylated KSA, and to produce KSA precursors, such as nascent KSA, and to produce subfragments of KSA. The recombinant-produced KSA is useful for the diagnosis, prognosis and treatment of disease states including adenocarcinomas of the lung, prostate, breast, ovary and colon/rectum; and for the creation of novel antibodies for treatment or diagnosis of the above.

Abstract: In a process for the production of a soluble native protein, such as immunoglobulin or methionine-prochymosin, in which an insoluble form of the protein is produced by a host organism transformed with a vector including a gene coding for the protein, the insoluble form of the protein is reversibly denatured in an alkaline aqueous solution at a pH selected to promote dissociation of a group or groups of the protein involved in maintaining the conformation of the protein, and the protein is subsequently allowed to renature by reducing the pH of the solution below a pH effective to denature the protein to produce the soluble native form of the protein. The pH of the alkaline aqueous is suitably in the range 9.0 to 11.5.

Abstract: This invention provides catalytic molecules capable of cleaving target nucleotide sequences. More specifically, the invention provides an endonuclease having nucleotide sequences which are of sufficient length to allow hybridisation to a target nucleotide sequence desired to be cleaved. The endonuclease contains a catalytic region comprising ribonucleotides and/or deoxyribonucleotides, or derivatives thereof which act to cleave a phosphodiester bond of the substrate nucleotide sequence. The catalytic region comprises nucleotides or derivatives thereof which are linked by linking groups which may comprise ribonucleotides, deoxyribonucleotides or combinations thereof.The endonucleases of the invention are useful in the cleavage of target RNAs associated with disease in humans and animals and in the inactivation of RNA transcripts in eukaryotic and prokaryotic cells, as well as the cleavage of RNA transcripts in-vitro.

Abstract: The invention relates to a signal peptide of the formula:MXKSTLLLLFLLLCLPSWNAGAand X represents a direct bond between M and K, an amino acid selected from the group comprising the 20 amino acids of the genetic code, or a peptide containing 2, 3 or 4 amino acids selected, each independently of the other, from the group comprising the 20 amino acids of the genetic code.