Abstract: The present invention provides compositions and methods for utilizing recombinant alphavirus vectors. Also disclosed are compositions and methods for making and utilizing eukaryotic layered vector initiation systems.

Abstract: T cell receptors (TCRS) that have higher affinity for a ligand than wild type TCRs are provided. These high affinity TCRs are formed by mutagenizing a T cell receptor protein coding sequence to generate a variegated population of mutants of the T cell receptor protein coding sequence; transforming the T cell receptor mutant coding sequence into yeast cells; inducing expression of the T cell receptor mutant coding sequence on the surface of yeast cells; and selecting those cells expressing T cell receptor mutants that have higher affinity for the peptide/MHC ligand than the wild type T cell receptor protein. The high affinity TCRs can be used in place of an antibody or single chain antibody.

Abstract: This invention describes the identification of pre-messenger RNA processing as a novel target of environmental stress caused for example by lithium and sodium toxicity. Overexpression of different types of proteins (or protein fragments) from different organisms but all involved in pre-mRNA processing, protects yeast from salt stress, which indicates that any stimulation of this process, independently of its mechanism, may counteract the toxic effects of mineral salts. A similar phenotype of tolerance to NaCl and to LiCl has been observed by overexpression of these types of proteins in transgenic Arabidopsis plants, demonstrating the generality of this protective effect in eukaryotic cells and organisms.

Abstract: Disclosed is a method for promoting auto-induction of transcription of cloned DNA in cultures of bacterial cells grown batchwise, the transcription being under the control of a promoter whose activity can be induced by an exogenous inducer whose ability to induce said promoter is dependent on the metabolic state of said bacterial cells. Initially, a culture media is provided which includes: i) an inducer that causes induction of transcription from said promoter in said bacterial cells; and ii) a metabolite that prevents induction by said inducer, the concentration of said metabolite being adjusted so as to substantially preclude induction by said inducer in the early stages of growth of the bacterial culture, but such that said metabolite is depleted to a level that allows induction by said inducer at a later stage of growth. The culture medium is inoculated with a bacterial inoculum, the inoculum comprising bacterial cells containing cloned DNA, the transcription of which is induced by said inducer.

Abstract: The present invention relates to the delivery of biologically active molecules, such as peptides, into the interior of cells by administering to the cells a complex comprising the molecule linked to an importation competent signal peptide. Such delivery can be utilized, for example, to treat and/or prevent inflammatory conditions, e.g., but not limited to, systemic inflammatory reactions such as endotoxic shock, localized inflammatory reactions such as inflammatory skin diseases and conditions, and inflammatory diseases such as autoimmune diseases.

Abstract: The present invention relates to a surface expression vector having pgsBCA, a gene coding poly-gamma-glutamate synthetase and a method for expression of target protein at the surface of microorganism using the vector. The vector, in which foreign genes are inserted, transforms microorganisms and makes foreign proteins expressed stably on the surface of microorganisms.

Abstract: The present invention provides methods and compositions for the efficient and enhanced secretion of a protein of interest from a host cell. In specific, proteins are secreted through the Sec-dependent pathway, involving the spoIIIJ and/or yqjG gene product(s). In some embodiments, expression of the spoIIIJ and/or yqjG gene product(s) is modulated by a promoter operably linked to the gene.

Abstract: Isolated DNAs which are DNAs having a base sequence represented by any of SEQ ID NOS:1 to 6 in Sequence Listing or fragments thereof and showing a stationary phase-specific promoter activity in gram positive bacteria.

Abstract: The present invention is directed to secreted and transmembrane polypeptides and to nucleic acid molecules encoding those polypeptides. Also provided herein are vectors and host cells comprising those nucleic acid sequences, chimeric polypeptide molecules comprising the polypeptides of the present invention fused to heterologous polypeptide sequences, antibodies which bind to the polypeptides of the present invention and to methods for producing the polypeptides of the present invention.

Abstract: In this application is described the expression and purification of a recombinant Plasmodium falciparum (3D7) LSA-NRC polypeptide. The method of the present invention produces a highly purified polypeptide which is useful as a vaccine and as a diagnostic reagent.

Abstract: This invention provides a modified vaccinia topoisomerase enzyme containing an affinity tag which is capable of facilitating purification of protein-DNA complexes away from unbound DNA. This invention further provides a modified sequence specific topoisomerase enzyme. This invention provides a method of ligating duplex DNAs, a method of molecular cloning of DNA, a method of synthesizing polynucleotides, and a method of gene targeting. Lastly, this invention provides a recombinant DNA molecule composed of segments of DNA which have been joined ex vivo by the use of a sequence specific topoisomerase and which has the capacity to transform a suitable host cell comprising a DNA sequence encoding polypeptide activity.

Abstract: The present invention provides an animal cell expressing a gene coding a ligand-responsive transcription control factor and securely maintaining a DNA comprising in a molecule, the following genes (a) and (b): (a) a reporter gene connected downstream from a transcription control region, in which said transcription control region substantially consists of a recognition sequence of said ligand-responsive transcription control factor and a minimum promoter which can function in said cell; and (b) a selective marker gene which can function in said cell; provided that the following gene (c): (c) a reporter gene connected downstream from a promoter which transcription activity is unchanged by having said responsive transcription control factor contacted with a ligand of said ligand-responsive transcription control factor, said reporter gene (c) coding a protein which can be differentiated from the protein coded by said gene (a) is not present in said cell and the like.

Abstract: The present invention is directed to novel polypeptides and to nucleic acid molecules encoding those polypeptides. Also provided herein are vectors and host cells comprising those nucleic acid sequences, chimeric polypeptide molecules comprising the polypeptides of the present invention fused to heterologous polypeptide sequences, antibodies which bind to the polypeptides of the present invention and to methods for producing the polypeptides of the present invention.

Abstract: Recombinant yeast are provided having, in one aspect, multiple exogenous LDH genes integrated into the genome, while leaving native PDC genes intact. In a second aspect, recombinant yeast are provided having an exogenous LDH gene integrated into its genome at the locus of a native PDC gene, with deletion of the native PDC gene. The recombinant yeast are useful in fermentation process for producing lactic acid.

Abstract: An isolated and purified transgenic Saccharomyces cerevisiae yeast cell comprising a disrupted ACR3 gene and an isolated DNA sequence comprising a promoter operably linked to a nucleic acid molecule encoding yeast cadmium factor resistance protein Ycf1p, is provided, as well as uses of the transgenic yeast cell.

Abstract: Artificial chromosomes, including ACes, that have been engineered to contain available sites for site-specific, recombination-directed integration of DNA of interest are provided. These artificial chromosomes permit tractable, efficient, rational engineering of the chromosome for a variety of applications.

Abstract: Methods are disclosed herein for inducing homologous recombination in a host cell comprising a target nucleic acid, using a single-stranded nucleic acid molecule. The single-stranded nucleic acid molecule has a sufficient number of nucleotides homologous to the target nucleic acid to enable homologous recombination with the target nucleic acid. The host cell includes a de-repressible promoter operably linked to a nucleic acid encoding a single-stranded binding protein and is deficient for mismatch repair. Isolated host cells of use in this method are also disclosed.

Abstract: A selection system free of antibiotic resistance genes, which is based on the use of an araD gene as a selection marker carried on a vector which is inserted in a bacterial strain deficient of the araD gene. The araD gene from E. coli encodes the L-ribulose-5-phosphate-4-epimerase. A method of selecting the cells transformed with a plasmid, which contains the araD gene. The non-antibiotic selection marker makes the system suitable for producing therapeutics. The araD gene is not essential for growth of the host but manipulation of it affects the growth under certain selective conditions. Deletion of araD leads to accumulation of substance which is toxic to the host but not to humans. The araD gene is relatively small and therefore a small plasmid may be constructed, which requires less energy for replication, and leads to increased growth rate and yield.

Abstract: A Bacillus licheniformis mutant host cell derived from a parent B. licheniformis host cell, which mutant host cell is mutated in one or more gene(s) encoding one or more secreted polypeptide(s) which is at least 80% identical to one or more of the polypeptides shown in SEQ ID NO's: 2 to 248, wherein the mutant host cell secretes at least 5% less of the one or more secreted polypeptide(s) than the parent host cell, when they are cultivated under comparable conditions.

Abstract: Disclosed are methods for obtaining expression of polypeptides in organisms employing alternative codon systems, and polynucleotides for use therein.