Abstract: Methods for the recombinant-DNA mediated production of xanthan gum and gum variants structurally related to xanthan are disclosed. The methods in part involve the synthesis of these polysaccharides in anaerobic and/or denitrifying hosts.In particular, plasmids pX209 and pRK290-H366 are disclosed which contain the genes, isolated from X. campestris, encoding Transferase I, Transferases II, Transferase III, Transferase IV, Transferase V, Ketalase, Acetylase and Polymerase. These plasmids have been deposited in the American Type Culture Collection under Accession Nos. 67051 and 67049, respectively.

Abstract: DNA regulatory elements that control cholesterol 7.alpha.-hydroxylase expression are disclosed. A gene construct is provided comprising at least one regulatory element and a reporter gene is used in an assay to detect a compound that modulates cholesterol 7.alpha.-hydroxylase enzyme regulation. Thus, a method for screening compounds that inhibit or stimulate expression of the enzyme is provided, as well as a method for detecting and isolating the transcription factors of the cholesterol 7.alpha.-hydroxylase gene.

Abstract: A transient expression system is disclosed that utilizes bacteriophage RNA polymerase in the presence of a DNA-based cytoplasmic virus to facilitate expression of a foreign gene in the cytoplasm of a eukaryotic cell.A method of expressing a foreign gene in the cytoplasm of a eukaryotic cell is also disclosed which comprises incorporating into the cytoplasm a DNA-based cytoplasmic virus, a suitable carrier comprising a gene for an RNA polymerase which gene is foreign to the carrier and to the cells, and a suitable carrier comprising a functional, cistron including a foreign gene flanked by a promotor sequence which is recognized by the RNA polymerase.

Abstract: Disclosed are novel DNA segments, vectors and plasmids containing multiple promoters for use with various polymerases in order to transcribe cloned DNA into RNA. A preferred vector, termed pTRIPLEscript.TM., is described which contains the SP6, T7, and T3 phage promoters in the same orientation and on the same side of a multiple cloning site. This vector efficiently synthesizes in vitro transcripts from all three promoters under conditions of both limiting and saturating nucleotide concentrations. This vector also promotes transcription without crosstalk, i.e., without nonspecific initiation at inappropriate promoters.

Abstract: A DNA cassette containing sequences encoding a lytic peptide under the transcriptional control of an immune system regulating sequence is disclosed. The control sequences normally permit expression only when a defined indicator(s) reflective of a disease state is present, and tightly inhibit expression when such indicator(s) is absent. The invention further extends to transgenic unicellular and multicellular organisms having the cassette stably integrated in their genetic material. A preferred embodiment of the cassette has the Shiva-1 lytic peptide under the control of IL-2 (interleukin-2) regulating sequences. In a preferred embodiment of multicellular organisms, the cassette is incorporated into the genomes of the germ cells and the organisms are capable of transmitting it to their offspring.

Abstract: The invention concerns a vaccine comprising an attenuated Salmonella bacterium which contains a nirB promoter operably linked to a DNA sequence encoding a heterologous protein. The nirB promoter directs expression of the heterologous protein in a host it is wished to vaccinate.

Abstract: A gene sequence that encodes the human CCAAT/enhancer binding protein ("C/EBP"), and recombinant vectors that are capable of mediating the expression of the C/EBP gene are described. The gene sequence and vector can be used in methods of gene therapy to treat cancer and other diseases.

Abstract: This invention relates to the use of promoters for ribonucleic acid amplification and other genetic manipulations. Processes are provided wherein complementary deoxyribonucleic acid (cDNA) is synthesized from a ribonucleic acid (RNA) sequence using a complementary primer linked to an RNA polymerase promoter region complement and then anti-sense RNA (aRNA) is transcribed from the cDNA by introducing an RNA polymerase capable of binding to the promoter region. Additional processes using the resulting aRNA are also described.

Abstract: Recombinant chimeric human rhinovirus and method for stimulation of a specific immune response. Design considerations, methods, and examples are described. Chimeric rhinoviruses can be used as vaccines and for a variety of other immunotechnological applications.

Abstract: A novel strain of Saccharomyces cerevisiae is useful as a host cell in the production of recombinant proteins. The novel S. cerevisiae cells transformed with a recombinant expression vector encoding a desired heterologous protein, preferably fused to a suitable N-terminal signal peptide, are cultivated under conditions that promote expression of the protein. Also provided are signal peptides derived by replacing the native signal peptidase cleavage site of a type I interleukin-1 receptor signal peptide with the tripeptide AlaXAla, wherein X represents an amino acid selected from Leu, Phe, and Gln. An expression system comprises a yeast host cell (preferably the novel S. cerevisiae strain) transformed with an expression vector comprising a promoter functional in yeast cells operably linked to DNA encoding the novel signal peptide, which is fused to the N-terminus of DNA encoding a desired heterologous protein.

Abstract: An improved vector upon introduction into a suitable host containing the thermolabile repressor C.sub.I renders the host capable of effecting expression of a desired gene. The vector is a double-stranded DNA molecule which includes in 5' to 3' order the following: the promoter and operator P.sub.L O.sub.L from lambda bacteriophage; the N utilization site; a first restriction enzyme site permitting replacement of the ribosomal binding site which follows thereafter; a ribosomal binding site; an ATG initiation codon or DNA which is converted into an ATG initiation codon upon insertion of the desired gene into the vector; a second restriction enzyme site for inserting the gene in phase with the ATG codon; a T.sub.1 T.sub.2 rRNA transcription termination sequence; an origin of replication and a gene associated with a selectable or identifiable phenotypic trait manifested when the vector is present in the host. The distance between the 3' end of the P.sub.L O.sub.

Abstract: A novel expression system using the heat-inducible bovine hsp70A promoter and associated cis-acting elements is disclosed. The system provides for the continuous production of a highly pure, authentic protein, substantially free of infectious viral and cellular protein contaminants.

Abstract: Sequence-specific oligonucleotides are provided having substantially pure chiral Sp phosphorothioate, chiral Rp phosphorothioate, chiral Sp alkylphosphonate, chiral Rp alkylphosphonate, chiral Sp phosphoamidate, chiral Rp phosphoamidate, chiral Sp phosphotriester, and chiral Rp phosphotriester linkages. The novel oligonucleotides are prepared via a stereospecific SN.sub.2 nucleophilic attack of a phosphodiester, phosphorothioate, phosphoramidate, phosphotriester or alkylphosphonate anion on the 3' position of a xylonucleotide. The reaction proceeds via inversion at the 3' position of the xylo reactant species, resulting in the incorporation of phosphodiester, phosphorothioate, phosphoramidate, phosphotriester or alkylphosphonate linked ribofuranosyl sugar moieties into the oligonucleotide.

Abstract: The present invention provides a method of detecting the presence of a nucleotide sequence within a double-stranded DNA in a sample comprising: a. digesting the double-stranded DNA with an exonuclease which converts at least a portion of the double-stranded DNA to single-stranded DNA, b. binding the single-stranded DNA with a nucleic acid probe which selectively hybridizes with the single-stranded DNA, and c. detecting hybridization between the single-stranded DNA and the nucleic acid probe, the existence of hybridization indicating the presence of the nucleotide sequence within the double-stranded DNA in the sample. The present invention further provides a method of detecting the presence of a nucleotide sequence in a sample comprising DNA which is the product of a DNA amplification technique. The invention also provides methods of sequencing and cloning using exonuclease.

Abstract: The present invention relates to methods and compositions involving an improved recombinant transfer vector for introducing a DNA sequence, encoding a recombinant protein, into an adenovirus genome in generating recombinant adenovirus. Recombinant protein production, in cells infected with the recombinant adenovirus, can approach levels as high as 15-20% of total cellular proteins. The improved transfer vector includes an expression cassette comprising sequentially a transcription promoter, a high efficiency leader, at least one splicing signal, an enhancer-like sequence, a cloning site and a plurality of polyadenylation sites.

Abstract: Methods for the enzymatic reduction and/or elimination of contaminant DNA from enzymatic amplification procedures are disclosed. These methods include the use of restriction endonucleases, DNAases and exonucleases which act upon contaminant DNA in polymerase chain reaction mixtures such that contaminant DNA is not amplified.

Abstract: The invention relates to a transposon consisting of a fragment of the R plasmid pCxM82B from Corynebacterium xerosis DSM 5021, to mobilizable, non-self-transferrable vectors which contain this transposon, and to methods for the mutagenesis of gram-positive bacteria by means of the transfer of these vectors and the generation of auxotrophic mutants.

Abstract: A DNA signal sequence initially isolated from Streptomyces griseus encodes a signal peptide which directs the secretion, via a fused intermediate, of a protein from the cell within which the DNA signal sequence is expressed. The signal sequence is derived from genes encoding protease A and protease B of S. griseus. The DNA signal sequence encodes a thirty-eight amino acid signal peptide. A DNA construct, including the DNA signal sequence and a gene sequence encoding a protein, when transformed into a living cell by a suitable vector, results in the signal peptide correctly directing the secretion of a mature protein of desired structure, particularly from prokaryotic genera selected for their ability to display enzymatic activity of a type typified by, but not exclusive to, that of protein disulphide oxidoreductase, EC 5.3.4.1, more particularly in the genera Streptomyces, and most particularly in Streptomyces lividans 66.

Abstract: A method for determining the efficacy of a therapeutic agent, in vitro, for a cancer expressing or overexpressing an oncogene product is described. The method is particularly useful for determining the efficacy of therapeutic agents that have a binding affinity for cancer that express HER-2/neu. N24, N28 and N29 monoclonal antibodies are described which have been identified by this method. One or more of these antibodies can be used as a therapeutic agent in the treatment of breast, stomach, ovarian or salivary cancers.

Abstract: The invention provided herein includes gram negative bacteria cells containing a gene encoding an enzyme with carbohydrate degrading activity that had been rendered competent to transformation. Carbohydrate degrading enzymes of interest for use in the invention include alpha-amylase. The competent cells of the subject invention may be frozen so as to provide for prolonged storage.Other aspects of the invention include methods for rendering gram negative bacterial cells, such as E. coli cells competent to transformation. These methods involve the step of transferring a gene encoding an enzyme with carbohydrate degrading activity into E. coli cells and subsequently rendering the cells competent using any of a variety of competency inducing procedures.