Abstract: The present invention relates to a method for improved ethanol production from xylose utilizing strain of Saccharomyces cerevisiae comprising genes for overexpression of xylose reductase, xylitol dehydrogenase and xylulokinase, wherein in addition to said genes, one or more specific genes of a defined group is overexpressed, and/or one or more genes of a defined group is deleted.

Abstract: The staling of an edible product made from dough can be retarded by adding a starch-degrading glucogenic exo-amylase of Family 13 to the dough, particularly an amylase from Thermotoga.

Abstract: Cystatin-based peptide tags, referred to here as inclusion body tags (IBTs), are disclosed useful for the generation of insoluble fusion peptides. The fusion peptides comprise at least one inclusion body tag operably linked to a peptide of interest. Expression of the fusion peptide in a host cell results in a product that is insoluble and contained within inclusion bodies in the cell and/or cell lysate. The inclusion bodies may then be purified and the protein of interest may be isolated after cleavage from the inclusion body tag.

Abstract: The present invention relates to novel phytases, in particular, of fungal origin, and also to their respective methods of production. The present invention relates more particularly to novel phytases derived from fungi of the Penicillium genus, in particular of the Penicillium sp. CBS 109899 strain, and also to the polynucleotides encoding these phytases. The invention also relates to vectors containing the polynucleotides, and to transformed host organisms expressing the phytases.

Abstract: The invention provides methods and materials related to the production of organic products such as glucuronic acid, ascorbic acid, and glucaric acid. Specifically, the invention provides cells, methods for culturing cells, isolated nucleic acid molecules, and methods and materials for producing various organic products such as glucuronic acid, ascorbic acid, and glucaric acid.

Abstract: The present invention relates to a ?5 desaturase, which has the ability to convert dihomo-?-linolenic acid (DGLA; 20:3 ?-6) to arachidonic acid (ARA; 20:4 ?-6) and/or eicosatetraenoic acid (ETA; 20:4 ?-3) to eicosapentaenoic acid (EPA; 20:5 ?-3). Isolated nucleic acid fragments and recombinant constructs comprising such fragments encoding ?5 desaturase along with a method of making long chain polyunsaturated fatty acids (PUFAs) using this ?5 desaturase in oleaginous yeast are disclosed.

Abstract: A gene encoding glutathione synthetase from Candida utilis is provided and food containing ?-glutamylcysteine cysteine or cystenylglycine is produced by cultivating Candida utilis modified by means of a gene encoding glutathione synthetase under a suitable condition and mixing the obtained culture or a fraction thereof or the culture or a fraction thereof subjected to heat treatment with a raw material of food or drink to process food or drink.

Abstract: The Invention pertains to methods, kits, molecules and cells to increase the rate or recombination and/or target recombination in dividing cells. In a particular aspect, the invention concerns methods and kits to induce targeted meiotic recombination.

Abstract: The invention provides recombinant cells that have been engineered such that ligand stimulation of a receptor expressed by the cells leads to amplified signal transduction responses. In one embodiment, the receptor-expressing cells have been engineered to carry a heterologous DNA construct comprising a gene encoding a protein that activates the signal transduction pathway, which gene is operatively linked to a promoter that is responsive to activation of the signal transduction pathway. Stimulation of the receptor by a ligand leads to expression of the heterologous DNA construct encoding the protein that activates the signal transduction pathway such that signals generated by ligand binding to the receptor are amplified. Preferred cells are yeast cells expressing heterologous G protein coupled receptors functionally coupled to the yeast pheromone response pathway and overexpressing Ste5p, Ste4p, Ste12p, Ste11p or a dominant truncation allele of Ste20 via a pheromone-responsive promoter.

Abstract: This invention pertains to nucleic acid fragments encoding plant proteins that are homologs to the cis-prenyltransferases UPP synthase from the bacterium Micrococcus luteus or Dedol-PP synthase from yeast Saccharomyces cerevisiae. More specifically, this invention pertains to cis-prenyltransferase homologs from wheat, grape, soybean, rice, African daisy, rubber tree latex and pot marigold.

Abstract: Two acyltransferases are provided, suitable for use in the manufacture of microbial oils enriched in omega fatty acids in oleaginous yeast (e.g., Yarrowia lipolytica). Specifically, the genes encoding phophatidylcholine-diacylglycerol acyltransferase (PDAT) and diacylglycerol acyltransferase (DGAT2) have been isolated from Y. lipolytica. These genes encode enzymes that participate in the terminal step in oil biosynthesis in yeast. Each is expected to play a key role in altering the quantity of polyunsaturated fatty acids produced in oils of oleaginous yeasts.

Abstract: The present invention relates to the elimination of mannosylphosphorylation on the glycans of glycoproteins in the yeast genus Pichia. The elimination of mannosylphosphorylated glycoproteins results from the disruption of the PNO1 gene and the newly isolated P. pastoris MNN4B gene. The present invention further relates to methods for producing modified glycan structures in host cells that are free of glycan mannosylphosphorylation.

Abstract: The present invention relates to a method for preparing an ethanol producing, xylose utilizing strain of Saccharomyces cerevisiae comprising genes for overexpression of xylose reductase, xylitol dehydrogenase and xylulokinase, wherein in addition to said genes for production o phosphoacetyltransferase, and acetaldehyde dehydrogenase are introduced and optionally overexpressed.

Abstract: The present invention makes available a rapid, effective assay for screening and identifying pharmaceutically effective compounds that specifically interact with and modulate the activity of a cellular receptor or ion channel. The subject assay enables rapid screening of large numbers of polypeptides in a library to identifying those polypeptides which induce or antagonize receptor bioactivity. The subject assay is particularly amenable for identifying surrogate ligands for orphan receptors.

Abstract: Lysophosphatidic acid acyltransferase (LPAAT) participates in the second step of oil biosynthesis and is expected to play a key role in altering the quantity of long-chain polyunsaturated fatty acids produced in oils of oleaginous organisms. The present application provides a nucleic acid fragment (identified as “LPAAT2”) isolated from Mortierella alpina encoding a LPAAT homolog that is suitable for use in the manufacture of oils enriched in omega fatty acids in oleaginous organisms. Most desirably, the substrate specificity of the instant LPAAT2 will be particularly useful to enable accumulation of long-chain PUFAs having chain lengths equal to or greater than C20 in oleaginous yeast, such as Yarrowia lipolytica.

Abstract: A method of providing papillomavirus like particles which may be used for diagnostic purposes or for incorporation in a vaccine for use in relation to infections causd by papillomavirus. The method includes an initial step of constructing one or more recombinant DNA molecules which each encode papillomavirus L1 protein or a combination of papillomavirus L1 protein and papillomavirus L2 protein followed by a further step of transfecting a suitable host cell with one or more of the recombinant DNA molecules so that virus like particles (VLPs) are produced within the cell after expression of the L1 or combination of L1 and L2 proteins. The VLPs are also claimed per se as well as vaccines incorporating the VLPs.

Abstract: Application of a functionalized porous membrane to purify nucleic acids, the binding properties of the membrane with respect to nucleic acids being adjustable by controlling the conditions of an ambient medium. The membrane is functionalized by deprotonated groups.

Abstract: Expression vectors and yeast cells that contain a heterologous G protein-coupled receptor gene and a gene mutation that causes increased sensitivity to receptor activation or a gene mutation that permits transcriptional activation of pheromone-responsive genes without cell cycle arrest. Methods of making the yeast cells.

Abstract: The present invention provides methods of introducing a polynucleotide into a target cell, wherein the method employs a light generating protein coding sequence acting as a reporter. An important advantage of the methods described herein is that drug resistant target cells or target cells having no useful auxotrophic markers can be effectively transformed. The present invention also includes transformed cells produced by the methods described herein. Also described are light generating protein coding sequence modifications, a variety of vectors, and methods of using the transformed cells of the present invention.

Abstract: A flocculent Saccharomyces cerevisae strain (BPSC-15) produces higher yields of ethanol in fermentations of fermentable sugar or fermentable starch/enzyme.

Abstract: The invention relates to a strain of the yeast Saccharomyces cerevisiae which, owing to deletion of the genomic sequences, no longer synthesizes hexose transporters and, as a consequence, can no longer grow on substrates with hexoses as the only carbon source, and whose ability of growing on a substrate with a hexose as the only carbon source is restored when it expresses a GLUT4 gene.

Abstract: Methods for preparing yeast with improved biotin productivity using integrating plasmids encoding biotin synthase. The yeast is transformed by an integrating plasmid, which includes a Candida utilis biotin synthase gene BIO2, an assistant DNA sequence to promote integration of the plasmid into the C. utilis genome, a promoter sequence, and a selection marker. Other embodiments include Saccharomyces cerevisiae integrating plasmids.

Abstract: A recombinant microbial cell comprises at least one increased expressible enzyme activity controlling anabolic metabolism of ammonia as a nutrient source, the increased enzyme activity being an NADH-dependent activity catalysing the reaction (a) 2-oxoglutarate+NH3+NADH?glutamate+NAD or being an activity catalysing the reaction (b) 2-oxoglutarate+glutamine+NADH?2 glutamate+NAD or being an activity catalysing the reaction (c) glutamate+NH3+ATP?glutamine+ADP+Pi. The increased enzyme activity is encoded by a nucleic acid coding sequence linked to an expression signal not natively associated with the nucleic acid coding sequence and is increased as compared to the expression of the enzyme activity when the nucleic acid coding sequence is associated with its native expression signal.

Abstract: This invention is directed to the transformation of the flavinogenic yeasts, Pichia guilliermondii and Candida famata, and mutants thereof, by electroporation (electrotransformation) and by spheroplast transformation. The invention is also directed to nucleic acid constructs such as vectors, plasmids, and ARS sequences which transform flavinogenic yeasts, and mutants thereof, at a high level and in a stable manner so as to result in stably transformed yeast host cells which express/produce recombinant products. This invention also is directed to flavinogenic yeasts, Pichia guilliermondii and Candida famata, and mutants and temperature sensitive mutants thereof, which produce or overproduce riboflavin.

Abstract: Disclosed herein are novel methods and materials for selecting transgenic cells. Specifically exemplified herein are positive selection methods that involve conferring to cells the ability to metabolize certain compounds, preferably arabitol, ribitol, raffinose, sucrose, mannitol or combinations thereof. Accordingly, transformed cells can be selected by simply subjecting them to a medium containing such compounds. The subject invention alleviates the disadvantages and concerns of negative selection methods, such as the unnecessary killing of transformed cells and the dispersal of potentially harmful genes (e.g., antibiotic or herbicide resistant genes) into the environment. Furthermore, novel nucleotide sequences relating to the E. coli rtl operon and arabitol dehydrogenase gene, and amino acid sequences relating to the gene products thereof are disclosed.

Abstract: This invention relates to functional gene arrays of yeast. Novel aspects include the individual yeast cells, methods for making the yeast and the arrays, the arrays and uses for the arrays. A diploid bearing special genetic properties has been constructed to facilitate cloning of heterologous genes capable of providing essential functions. A selection method, using this strain allows the identification haploid yeast strains dependent for life on heterologous essential genes. The arrays of these strains comprise a library of unique members where each member is dependent for survival on the function of a heterologous gene complementing a different essential host gene which has been inactivated by the insertion of a dominant selectable marker. These arrays provide screening platforms for agents that specifically target the activity of these heterologous genes.

Abstract: The present invention is directed to a recombinant vector for transforming yeast and a process for transforming yeast thereby, more particularly to a recombinant vector comprising a gene encoding a mutated L41 protein having cycloheximide-resistant activity and a ribosomal DNA. The recombinant vector and the process for transforming thereby of the present invention is applicable to the efficient and stable integration of desired foreign DNA into yeast genome, thus providing useful tools for the production of a natural pigment, astaxanthin.

Abstract: Yeast cells are mutagenized to obtain desirable mutants. Mutagenesis is mediated by a defective mismatch repair system which can be enhanced using conventional exogenously applied mutagens. Yeast cells with the defective mismatch repair system are hypermutable, but after selection of desired mutant yeast strains, they can be rendered genetically stable by restoring the mismatch repair system to proper functionality.

Abstract: This invention relates generally to the field of monitoring production of gene products. In particular, the invention provides methods of monitoring production of a gene product, methods of screening for modulators of production of a gene product, and methods of screening for cellular targets amenable to regulation by a treatment using a plurality of reporter gene systems. The methods described herein find uses in a number of fields such as drug discovery, agricultural or industrial production and environmental monitoring or protection.

Abstract: This invention involves a process for selecting and producing eukaryotic alkaline phosphatase in yeast. Yeast cells are subjected to multiple transformations using a vector comprising a first resistance marker gene and the alkaline phosphatase gene. Those strains that grow in media containing the first resistance marker are further transformed using a vector comprising a second selection marker gene and the alkaline phosphatase gene. Transformants that grow in media containing the second selection marker are selected for expressing the eukaryotic alkaline phosphatase.

Abstract: Disclosed is a transformed yeast cell containing a first heterologous DNA sequence which codes for a mammalian G protean coupled receptor and a second heterologous DNA sequence which codes for a mammalian G protein ? subunit (mammalian G?). The first and second heterologous DNA sequences are capable of expression in the cell, but the cell is incapable of expressing an endogenous G protein ?-subunit (yeast G?). The cells are useful for screening compounds which affect the rate of dissociation of G? from G?? in a cell. Also disclosed is a novel DNA expression vector useful for making cells as described above. The vector contains a first segment comprising at least a fragment of the extreme amino-terminal coding sequence of a yeast G protein coupled receptor. A second segment is positioned downstream from the first segment (and in correct reading frame therewith), with the second segment comprising a DNA sequence encoding a heterologous G protein coupled receptor.

Abstract: Disclosed herein are compositions and methods which are useful in the identification and isolation of components involved in transmembrane receptor-mediated signaling. Such components include the receptors themselves (e.g., tyrosine kinase receptors, cytokine receptors and tyrosine phosphatase receptors), as well as ligands which bind the receptors and modulators of the downstream intracellular catalytic event which characterizes receptor-mediated signalling. Two novel ligands for the FGF receptor and the nucleotide sequences encoding them are also described.

Abstract: The present invention relates to methods of obtaining a mutant cell from a filamentous fungal parent cell, comprising: (a) obtaining mutant cells of the parent cell; (b) identifying the mutant cell which exhibits a more restricted colonial phenotype and/or a more extensive hyphal branching than the parent cell; and (c) identifying the mutant cell which has an improved property for production of a heterologous polypeptide than the parent cell, when the mutant and parent cells are cultured under the same conditions.

Abstract: The present invention provides methods of converting or increasing conversion of a fatty acid to its corresponding dicarboxylic acid. The methods comprise isolating a promoter from a yeast gene which gene is induced when the yeast is grown on a fatty acid or alkane substrate, and operably linking the promoter to a gene involved in dicarboxylic acid production to form an expression vector. Yeast cells are subsequently transformed with such an expression vector and cultured in a media containing an organic substrate biooxidizable to a mono- or polycarboxylic acid, and resultant yeast cells convert or increase conversion of fatty acids to their corresponding dicarboxylic acids. Examples of promoters that may be used in the methods of the present invention include those from C. tropicalis catalase, citrate synthase, 3-ketoacyl-CoA thiolase A, citrate synthase, O-acetylhomserine sulphydrylase, protease, carnitine O-acetyltransferase, hydratase-dehydrogenase, and epimerase genes.

Abstract: Recombinant materials, including DNA constructs, expression vectors, and host cells, and methods of identifying an environmental stimulus or a gene that alters the lifespan of an organism are identified. The recombinant DNA constructs include first and second chimeric genes that both encode substantially the same protein that is required for yeast replication, the first chimeric gene containing a promoter responsive to growth medium conditions and the second chimeric gene containing a promoter operable in mother cells but not daughter cells.

Abstract: The yeast which has &ggr;-glutamylcysteine-producing ability and is auxotrophic for pantothenic acid is proliferated in a medium containing a sufficient amount of pantothenic acid, and then it is cultured in a medium containing a limited amount of pantothenic acid to increase the &ggr;-glutamylcysteine content in its cells, whereby the yeast in which &ggr;-glutamylcysteine is accumulated is obtained.

Abstract: The present invention relates to methods for producing a polypeptide, comprising: (a) cultivating a fungal host cell in a medium conducive for the production of the polypeptide, wherein the fungal host cell comprises a nucleic acid construct comprising a first nucleotide sequence encoding a signal peptide operably linked to a second nucleotide sequence encoding the polypeptide, wherein the first nucleotide sequence is foreign to the second nucleotide sequence and the 3′ end of the first nucleotide sequence is immediately upstream of the initiator codon of the second nucleotide sequence. The present invention also relates to the isolated signal peptide sequences and to constructs, vectors, and fungal host cells comprising the signal peptide sequences operably linked to nucleotide sequences encoding polypeptides.

Abstract: A method for detecting protein-protein interactions is provided, in which two fusion proteins are prepared and allowed to interact with each other. The interaction between the two fusion proteins leads to protein trans-splicing, generating an active and detectable reporter.

Abstract: A process for the production of ergosterol and its intermediate products using recombinant yeasts and plasmids for transformation of yeasts is described.

Abstract: The present invention provides a simple method for splitting and loss of a chromosome in yeast. The method for modifying a chromosome in yeast includes preparing a linear chromosome splitting vector (1) having a target sequence (a), a marker gene sequence and (C4A2)n sequence in this order; preparing a linear chromosome splitting vector (2) having a target sequence (b), a centromere sequence of a yeast chromosome and (C4A2)n sequence in this order; and introducing the chromosome splitting vectors (1) and (2) into yeast. Herein, n is each independently an integer of 6 to 10. Although this chromosome splitting vector has a repetitive sequence of 5′-CCCCAA-3′, it can be amplified specifically with PCR, so that a chromosome splitting vector can be prepared significantly simply and easily, compared with the conventional DNA splitting method.

Abstract: The present invention relates to a method of screening for constitutively active mutants of a desired eukaryotic MAPK pathway member of a MAPK pathway member, comprising the steps of (a) providing a mutant yeast strain devoid of an upstream kinase; (b) providing a DNA library of different mutants of a mutagenized gene coding for the desired MAPK pathway member; (c) introducing said library into said yeast strain under conditions suitable for activation of the yeast MAPK pathway; (d) detecting an end-point indication for activation of the yeast pathway; and (e) optionally isolating said constitutively activated mutant from selected rescued clones. The invention further relates to isolated constitutively active mutants of the MAPK pathway member, and their various uses particularly, in a method of screening for substances which are inhibitors of a MAPK pathway, and in drug design.

Abstract: The invention relates to a method for quantitation of recombinant proteins of interest in the absence of specific antibodies, which is particularly suitable for screening cell clones expressing heterologous genes.

Abstract: A method for generating a Drosophila clipped FRT (cFRT) chromosome is provided, wherein the chromosome is insensitive to a P transposase but remains functional to a yeast site-specific flippase recombinase (FLP). The method includes steps of: (a) exposing a FRT chromosome to the P transposase for occurring a local and imprecise transposition, wherein the FRT chromosome contains a P[FRT] insertion with a selection marker gene, (b) screening the P[FRT] insertion insensitive to the P transposase to obtain screened products, (c) selecting candidate products from the screened products by further examinations, and (d) exposing the candidate products by the P transposase and selecting a desired product by the further examinations to obtain the Drosophila clipped FRT (cFRT) chromosome insensitive to the P transposase but remaining functional to the yeast site-specific flippase recombinase. The cFRT2L2R chromosome can be used as the direct target in the direct P-transposon-induced mutagenesis.

Abstract: The present invention provides genetically engineered strains of Pichia capable of producing proteins with reduced glycosylation. In particular, the genetically engineered strains of the present invention are capable of expressing either or both of an (&agr;-1,2-mannosidase and glucosidase II. The genetically engineered strains of the present invention can be further modified such that the OCH1 gene is disrupted. Methods of producing glycoproteins with reduced glycosylation using such genetically engineered stains of Pichia are also provided.

Abstract: The present invention relates to yeast cells containing the SRB1/PSA1 gene and/or the PKC1 gene or functional derivatives thereof operatively linked to a heterologous inducible promoter and also to methods of regulating yeast cell lysis and flocculation and methods of fermentation using such yeast cells.

Abstract: The invention provides isolated nucleic acid and amino acid sequences of Candida albicans cofilin, methods of screening for modulators of such protein, and kits therefore.

Abstract: Methods for making proteins with anti-angiogenic properties are disclosed. The system used is a yeast expression system which produces biologically active proteins at high titer.

Abstract: The invention provides methods and materials related to the production of organic products such as glucuronic acid, ascorbic acid, and glucaric acid. Specifically, the invention provides cells, methods for culturing cells, isolated nucleic acid molecules, and methods and materials for producing various organic products such as glucuronic acid, ascorbic acid, and glucaric acid.

Abstract: Described herein are methods for increasing the amount of protein secreted by a cell. In one case, a cell is provided which contains a heterologous nucleic acid encoding a protein having unfolded protein response modulating activity and a heterologous nucleic acid encoding a protein of interest to be secreted. In one case, the protein having unfolded protein response modulating activity is selected from the proteins selected from the group consisting of HAC1, PTC2 and IRE1. The protein of interest can be any secreted protein such as a therapeutic or an industrial enzyme. For example the protein can be selected from the group consisting of lipase, cellulase, endo-glucosidase H, protease, carbohydrase, reductase, oxidase, isomerase, transferase, kinase, phosphatase, alpha-amylase, glucoamylase, lignocellulose hemicellulase, pectinase and ligninase.

Abstract: A method for detecting interactions between first and second interacting molecules a variable sensitivity. This variable sensitivity may be obtained by providing for the overexpression of either a bait hybrid protein containing a DNA binding domain (desensitization) or a prey hybrid protein containing the DNA activation domain for a reporter gene (enhanced sensitivity). The use of exogenous activators of one or the other according to the needs of a particular system is readily accomplished.