Abstract: This invention relates to the preparation and use of expression systems capable of producing heterologous polypeptides in halobacterial hosts.

Abstract: The present invention is directed to a recombinant vector capable of directing the synthesis of gas vesicles in a non-floating cells. Cells transformed by the vector of the invention are also provided. The present invention is further directed to a method for conferring buoyancy to non-floating cells. The method of the instant invention is useful in harvesting cells, for example in fermentation processes, and in bioremediation, for example in conferring buoyancy to hydrocarbon-degrading bacteria.

Abstract: NIb replicase gene of papaya ringspot virus replicase strain FLA.83 W is provided.

Abstract: This invention is directed to L5 shuttle phasmids capable of delivering foreign DNA into mycobacteria and to methods of producing L5 shuttle phasmids. In addition, this invention is directed to a method of generating mycobacterial mutations and to a method of producing mycobacterial vaccines.

Abstract: A method of transient transfection of obligate intracellular parasites is described. This method is exploited to develop a system for stable transformation utilizing selectable genes. For example, introduction of the chloramphenicol acetyl transferase (CAT) gene fused to Toxoplasma flanking sequences followed by chloramphenicol selection results in parasites stably expressing CAT. DNA hybridization analysis indicated that the CAT gene had inserted via homologous recombination.

Abstract: Novel polypeptide compositions based on the amino acid sequence of tissue plasminogen activator (tPA) are provided having improved properties over natural tissue plasminogen activator. Particularly, enhanced specific activity, reduced response to inhibition by plasminogen activator inhibitor, fibrin stimulation of plasminogenolytic activity and/or enhanced affinity to fibrin surfaces are provided by modifying one or more loci by deletions or substitutions. One or both of the N- or C-termini may be modified.

Abstract: The present invention is directed to a process for the production of a peptide, polypeptide, or protein having a predetermined property. In accordance with one embodiment, the process begins by producing by way of synthetic polynucleotide coupling, stochastically generated polynucleotide sequences. A library of expression vectors containing such stochastically generated polynucleotide sequences is formed. Next, host cells containing the vectors are cultured so as to produce peptides, polypeptides, or proteins encoded by the stochastically generated polynucleotide sequences. Screening or selection is carried out on such host cells to identify a peptide, polypeptide, or protein produced by the host cells which has the predetermined property. The stochastically generated polynucleotide sequence which encodes the identified peptide, polypeptide, or protein is then isolated and used to produce the peptide, polypeptide, or protein having the predetermined property.

Abstract: A system is described which utilizes a novel system of repressor titration for maintenance of a plasmid useful in gene therapy and production of a recombinant protein. The system utilizes a transformed host cell containing a plasmid including an operator susceptible to binding by a repressor expressed in trans, a first chromosomal gene encoding the repressor, and a second chromosomal gene that is functionally associated with an operator and essential for cell growth, wherein the plasmid is present in the cell in sufficient numbers to titrate the repressor such that the essential gene is expressed, thereby permitting cell growth.

Abstract: The invention concerns a DNA fragment located in front of the malate synthase gene of a coryne-form bacterium and isolated from the latter. Any structural gene which codes for a protein can be inserted after this DNA fragment. After transformation of such a construct into a coryne-form bacterium, expression of the structural gene inserted after the DNA fragment is regulated. The invention also concerns a process for synthesizing any protein by culturing a transformed coryne-form bacterium. A bacterium of this type contains in replicable form a DNA fragment isolated from the malate synthase gene of a coryne-form bacterium, and after which the structural gene which codes for the protein to be synthesized is inserted. Since expression of the structural gene which codes for the protein to be synthesized is regulated by the DNA located in front of it, the structural gene is expressed and the desired protein synthesized as soon as a suitable inducing agent is added to the medium.

Abstract: An enzyme is altered by introducing an additional hydrophobic group to enhance the hydrophobic environment in the enzyme so as to interfere with entrance of a water molecule. In this connection, the site of introduction and kind of hydrophobic group to be introduced are selected on the basis of comprehensive analysis of amino acid sequence, three-dimensional structure, reaction mechanism or the like of the target enzyme. Using this method, an amino acid residue of neopullulanase derived from Bacillus stearothermophilus was replaced by another amino acid.

Abstract: A DNA construct wherein a DNA fragment which is recombinable in yeast chromosomal DNA is directly or indirectly linked at both ends of a DNA fragment which comprises a pair of R sensitive sequences oriented in the same direction and flaking both an R gene placed under the control of an inducible promoter and an expressible selective marker gene, which is a DNA construct designed with the R sensitive sequences non-symmetrically shortened, so that no functionable R sensitive sequence remains after the R sensitive sequence recombination has occurred by expression of the R gene and the selective marker has been removed. Since no functionable R sensitive sequence remains after removal of the selective marker, recombination does not occur again, and thus the same selective marker may be used for multiple insertions of foreign genes.

Abstract: An isolated SDS-resistant hyperthermostable .beta.-galactosidase gene derived from Pyrococcus furiosus. Examples thereof include genes respectively shown in SEQ ID NO: 1 and SEQ ID NO: 2 and genes each hybridizable with the above gene shown in SEQ ID NO: 1. A method of cloning the hyperthermostable .beta.-galactosidase gene in which each of the above genes or pacts thereof is used as a probe or primer. A process for producing a hyperthermostable .beta.-galactosidase by culturing a transformant into which a plasmid containing each of the above gene has been introduced.

Abstract: Novel polyketides and novel methods of efficiently producing both new and known polyketides, using recombinant technology, are disclosed. In particular, a novel host-vector system is described which is used to produce polyketide synthases which in turn catalyze the production of a variety of polyketides.

Abstract: Strains of Pseudomonas have been genetically engineered to have enhanced biocontrol properties. The strains of the invention are particularly effective against plant pathogenic fungi such as species of Rhizoctonia and Pythium, because the strains produce enhanced amounts of antifungal metabolites such as pyrrolnitrin that are active against these fungal pathogens. Both the genetically modified biocontrol strains and the antifungal metabolites can be used as active agents for biocontrol compositions.

Abstract: A system is used to express clostridial gene constructions in a clostridial host. A mobilizable transfer plasmid is described which permits the direct transfer of the plasmid, and genes carried on it, from E. coli into Clostridium species. A promoter is described for use in clostridial species. Also, a useful host strain is used which is nontoxigenic and which permits high levels of expression of clostridial genes using the clostridial promoter.

Abstract: This invention relates to drug binding proteins, to genes encoding same and to assays and methods for screening pharmaceuticals. More specifically, this invention relates to a Cytokine Suppressive Anti-Inflammatory Drug (CSAID) binding protein, to a gene encoding same and to assays and screens useful in the evaluation and characterization of drugs of this pharmacologic class.

Abstract: Escherichia coli plasmid vectors are provided which have a 5'-terminal untranslated region (inclusive of the promoter region and Shine-Dalgarno sequence) of the Escherichia coli lipoprotein gene, which region is improved to thereby enable direct production of useful polypeptides in substantially complete form.

Abstract: The invention provides customized proteases (i.e., mutant enzymes), methods of making customized proteases, as well as methods of using customized proteases. The customized proteases of the invention are derived from the known proteases. Altered transacylation reactions include the capability to perform transacylation reactions not substantially catalyzed by the known protease or the capability to perform transacylation reactions with improved yields, or both. The methods of the invention provide for customized proteases through site specific or random mutagenesis of the active site amino acids of the known proteases. The invention also provides for methods of using the customized proteases to prepare a preselected transacylation products. The preselected transacylation products produced can be modified by substitution at the N- or C-terminal with nucleophiles such as L-amino acids, D-amino acids, amino acid amides, and radioactive amino acids.

Abstract: Methods and a kit are provided for characterizing small molecules from a library of small molecules or alternatively identifying protein targets to which known small molecules bind. The methods include forming hybrid ligand in which at least one ligand is a small molecule. The hybrid ligand is introduced into cells that in turn contain a first and a second expression vector. Each expression vector includes DNA for expressing a hybrid protein that encodes a target protein linked to a coding sequence for a transcriptional module. The cells further contains a reporter gene, the expression of which is conditioned on the proximity of the first and second hybrid proteins to each other, an event that occurs only if the hybrid ligand binds to target sites on both hybrid proteins. Those cell which express the reporter gene are selected and the unknown small molecule or the unknown hybrid protein is identified.

Abstract: The present invention provides recombinant DNA vehicles which are suitable for the microbial expression of DNA encoding a heterologous polypeptide which comprises a portion of the trp operon having the promoter-operator and leader ribosome binding site, and a restriction site providing an insertion site for the DNA sequences encoding the heterologous polypeptide, wherein the restriction site is located 3' of the leader ribosome binding site as a substitute for the Taq I site of the trp promoter-operator and is selected from the group consisting of Xba I and Eco RI. Also provided are E. coli strains transformed with the above described recombinant DNA vehicles.

Abstract: The present invention is directed to recombinant plant viral nucleic acids and to hosts infected thereby. The recombinant plant viral nucleic acids comprise a native plant viral subgenomic promoter, at least one non-native plant viral subgenomic promoter, a plant viral coat protein coding sequence, and optionally, at least one non-native nucleic acid sequence to be transcribed or expressed in the infected host plant. The recombinant plant viral nucleic acids are stable, capable of systemic infection and capable of stable transcription or expression in the plant host of the non-native nucleic acid sequences.

Abstract: A novel uracil DNA glycosylase enzyme (referred to as Bpa UDG) has been identified in Bacillus pallidus and the gene encoding Bpa UDG has been cloned, sequenced and expressed to produce a recombinant UDG protein. The enzyme is thermostable and exhibits reaction kinetics similar to E. coli UDG. It is effectively inhibited by B. subtilis UGI.Bpa UDG may be used to inactivate contaminating amplicons in nucleic acid amplification reactions, particularly at higher reaction temperatures. It may also be used to generate Bpa UDG-specific antibodies for purification of Bpa-UDG or for detecting Bpa UDG in a sample. Certain Bpa UDG antibodies may inactivate the enzyme and may therefore be useful as substitutes for UGI or heat, or in combination with UGI and/or heat, for controlling UDG activity in a reaction.

Abstract: The invention provides a process of manufacture of a recombinant protein called Asp-Pallidipin. Asp-Pallidipin inhibits the collagen-induced platelet aggregation of mammalian platelets. The Asp-Pallidipin comprises(i) a protein (Pallidipin) selected from the group of Pallidipin proteins, and(ii) the amino acid aspartic acid,wherein the aspartic acid is connected by a peptide bond with the N-terminal end of Pallidipin.

Abstract: The invention relates to an improved process for the production of a secondary metabolite comprising (i) fermentation of a microorganism capable of producing said secondary metabolite, and (ii) recovering said metabolite in substantially pure form. Said microorganism has been modified in a manner whereby the expression of one or more of the DNA sequences coding for (a) peptide(s), (a) protein(s) or (an) enzyme(s), involved in or interfering with the biosynthetic pathway of said secondary metabolite, is regulated differently from the regulation of said DNA sequence(s) in the original microorganism. Further contemplated is a process for production of said microorganism, a DNA construct, a vector or transformation vehicle, a microorganism capable of producing secondary metabolite and finally a secondary metabolite product.

Abstract: This invention discloses a PACAP receptor protein or a salt thereof, a DNA comprising a DNA fragment coding for the protein, a method for preparing the protein, antibody against the protein and use of the protein, DNA and antibodies. A PACAP receptor protein was purified from the bovine cerebrum. DNAs coding for PACAP receptor proteins were isolated from bovine, rat and human cDNA libraries, and their nucleotide sequences were determined. The PACAP receptor proteins and the DNAs coding for the proteins of the present invention can be used for (1) acquisition of antibodies and antisera, (2) construction of expression systems of recombinant receptor proteins, (3) development of receptor binding assay systems using said expression systems and screening of potential compounds for drugs, (4) execution of drug design based on the comparison of ligands and receptors which are structurally similar to each other, (5) preparation of probes and PCR primers in gene diagnosis, and the like.