Abstract: The present invention is directed to expression vectors and yeast cells transformed therewith containing a first heterologous nucleotide sequence which codes for a G protein-coupled receptor, for example, the somatostatin receptor, and a second nucleotide sequence which codes for all or a portion of a G protein &agr;&bgr;&ggr; complex. Said heterologous protein is physically expressed in a host cell membrane in proper orientation for both stereoselective binding of ligands, as well as functional interaction with G proteins on the cytoplasmic side of the cell membrane. In some embodiments, a nucleotide sequence encoding a heterologous or chimeric G&agr; protein is expressed in conjunction with nucleotide sequences from the yeast G protein &bgr;&ggr; subunits. A second aspect of the present invention provides expression vectors and yeast cells transformed therewith encoding chimeric yeast/heterologous G protein coupled receptors.

Abstract: The present invention relates to growing and testing microorganisms in a multitest format which utilizes a gel forming matrix for the rapid screening of clinical and environmental cultures. The present invention is suited for the characterization of commonly encountered microorganisms (e.g., E. coli, S. aureus, etc.), as well as commercially and industrially important organisms from various and diverse environments (e.g., the present invention is particularly suited for the growth and characterization of the actinomycetes and fungi). The present invention is also particularly suited for comparative analysis of phenotypic differences between cell types, including strains of microorganisms that have been designated as the same genus and species, as well as other cell types (e.g., mammalian, insect, and plant cells).

Abstract: Described herein are methods for the production of monoclonal antibodies in filamentous fungi host cells. The monoclonal antibodies are expressed as full-length fusion proteins that retain functional antigen binding and antibody-dependent cellular cytotoxicity capabilities. Improvements in the cleavage of the glucoamylase-light chain fusion protein to yield a mature antibody are also provided. The antibodies produced in filamentous fungi show equivalent pharmacokinetic disposition to antibodies produced in mammalian cells.

Abstract: The present invention includes a method to detect IgE using a human Fc epsilon receptor (Fc&egr;R) to detect IgE antibodies in a biological sample from a cat, a dog, or a horse. The present invention also relates to kits to perform such methods.

Abstract: This invention relates to modified host cells which express heterologous fused proteins and methods of screening for test samples having peptide binding activity; wherein the modified host cell comprises: (a) a gene sequence encoding a heterologous fusion protein; the fusion protein comprising a first peptide of a peptide binding pair, or segment of the first peptide, which is joined to either a DNA binding domain or its corresponding transcriptional activation domain of a transcriptional activation protein; (b) a gene sequence encoding a heterologous fusion protein, the fusion protein comprising a second peptide of the peptide binding pair in (a), or a segment thereof, fused to either a DNA binding domain or its corresponding transcriptional activation domain, whichever one is not employed in (a); (c) a reporter gene operatively associated with the transcriptional activation protein, or a portion thereof; (d) optionally, a deletion or mutation in the chromosomal DNA of the host cell for the transcriptional a

Abstract: A method is described for the in vivo identification of epitopes of an intracellular antigen comprising the steps of :a) co-transforming of cells by a first vector including the nucleotide sequence encoding for the region of an antibody able to recognise and bind the intracellular antigen and by a second vector comprising the nucleotide sequence encoding for a peptide; b) growing co-transformed cells in such an environment that only cells wherein the antibody region and peptide recognise and interact each other are able to replicate and/or be recognised because: the antibody region able to recognise and bind the intracellular antigen is associated with a first molecule; the peptide is associated with a second molecule; the interaction of the first with the second molecule generates a selectable phenotype and/or recognisable signal; and the interaction of the first with the second molecule occurs only when the antibody region and peptide recognise and interact each-other; c) selecting the b) cells and identify

Abstract: The present invention provides antigenic peptides useful for the production of antibodies which selectively bind to an enzyme involved in the &ohgr;-oxidation of fatty acids and alkanes to &agr;,&ohgr;-dicarboxylic acids in yeast. Antibodies which specifically bind to an enzyme involved in the &ohgr;-oxidation of fatty acids and alkanes to &agr;,&ohgr;-dicarboxylic acids in yeast are also provided. In addition, methods of producing the subject antibodies, a method of detecting the presence and amount of a specific enzyme involved in the &ohgr;-oxidation of fatty acids and alkanes to &agr;,&ohgr;-dicarboxylic acids, and a method of monitoring the degree of enzyme induction and/or enzyme stability in a mixture during &ohgr;-oxidation of fatty acids and alkanes to &agr;,&ohgr;-dicarboxylic acids in yeast, are also provided.

Abstract: Compositions, methods, and kits are provided for efficiently generating and screening a library of highly diverse protein complexes for their ability to bind to other proteins or oligonucleotide sequences. In one aspect of the invention, a library of expression vectors is provided for expressing the library of protein complexes. The library comprises a first nucleotide sequence encoding a first polypeptide subunit; and a second nucleotide sequence encoding a second polypeptide subunit. The first and second nucleotide sequences each independently vary within the library of expression vectors. In addition, the first and second polypeptide subunit are expressed as separate proteins which self-assemble to form a protein complex, such as a double-chain antibody fragment (dcFv or Fab) and a fully assembled antibody, in cells into which the library of expression vectors are introduced.

Abstract: An assay to determine the specific expression and suppression of proteins in response to a stressor is disclosed. An organism exposed to a stressor, including disease caused by exposure to, e.g., a parasite, or a substance suspected of causing an adverse effect, is assayed to determine a first set of proteins expressed and a second set of proteins suppressed in response to the stressor. The amount of each protein expressed and the amount of each protein suppressed can be statistically analyzed to determine which proteins are most useful in diagnosing the stressor. A protein profile for a first stressor can be compared to protein profiles for a second stressor, a third stressor, etc. A distinct protein expression signature (PES) for the first stressor can be identified by determining subsets fo proteins expressed and/or suppressed only in response to the first stressor.

Abstract: The invention provides a method of characterizing a target that binds to a ligand. The method comprises providing ligands, optionally attached to a support, and contacting the ligands with targets to allow at least one target to bind to at least one ligand. The method further comprises immobilizing the resulting complexes in a first matrix, such that each complex has a different position within the first matrix, and transferring the target of the complex to a second matrix. The position of the target within the second matrix corresponds to the position of the ligand-support complex within the first matrix. The target on the second matrix is then detected.

Abstract: A biosensor includes a substrate with areas of active receptive material disposed thereon. The receptive material is specific for an analyte of interest. A pattern of the active areas is defined on the substrate by an oxidizing photo-masking process.

Abstract: Hemolysin isolated from hemolysin-producing fungi can be used to detect if a human or other animal has been exposed to a hemolysin-producing fungus. The method and proteins of the present invention can be used to screen humans and other animals for exposure to such fungi, as well as to produce vaccines for protecting humans and other animals that may be exposed to such fungi.

Abstract: A biosensor includes a substrate with a layer of receptive material disposed thereon. The receptive material is specific for an analyte of interest. A pattern of active and inactive areas of the receptive material are defined in the receptive material layer by a masking process.

Abstract: A biosensor includes a substrate with a layer of receptive material disposed thereon. The receptive material is specific for an analyte of interest. A pattern of active and inactive areas of the receptive material are defined in the receptive material layer by a masking process.

Abstract: A biosensor includes a substrate member with a pattern of active areas of receptive material and a pattern of blocking material layers. The receptive material and blocking material are attached to the substrate member with a photo-reactive crosslinking agent activated in a masking process. The receptive material is specific for an analyte of interest.

Abstract: A biosensor includes a substrate with a layer of receptive material disposed thereon. The receptive material is specific for an analyte of interest. A pattern of active and deactivated areas of the receptive material are defined in the receptive material layer by a masking process.

Abstract: Yeast cells are engineered to express both a surrogate of a pheromone system protein (e.g., enzymes involved in maturation of &agr;-factor, transporters of a-factor, pheromone receptors, etc.) and a potential peptide modulator of the surrogate, in such a manner that the inhibition or activation of the surrogate affects a screenable or selectable trait of the yeast cells. Various additional features improve the signal-to-noise ratio of the screening/selection system.

Abstract: The present invention relates to a composition useful for qualitative screening of phosphate solubilizing microorganism, and also relates to a synergistic composition for qualitative screening of phosphate solubilizing microorganisms, based upon visual observation.

Abstract: The present invention relates to procedures for the detection or for the determination of solid phase-associated factors, which are multiply associated with the same solid phase. According to the invention, the sample is brought into contact with a transmitter particle, on which at least one ligand having binding affinity for a solid phase-associated factor and a transmitter are immobilized, and a receiver particle, on which at least one ligand having binding affinity for said solid phase-associated factor and a receiver is immobilized, and then the signal is determined which results when transmitter and receiver are brought sufficiently close to one another. In particular, the invention relates to the detection of cell surface receptors which can be used for the typing of cells or for the determination of cell activation states. It is thus possible to replace the hitherto widely customary flow cytometry by a more simple procedure.

Abstract: The present invention relates to biocatalysts that are cells, optimally of the Crabtree-negative phenotype, comprising expression vectors encoding genes heterologous to the cell that enable increased production of organic products. More specifically, the invention relates to genetically modified Candida cells, methods for making the Candida cells, and their use in production of organic products, particularly lactic acid.

Abstract: Processes for preparing controlled samples of particles, including microorganisms and cells are described. A sample of particles is provided and separated into a predetermined number of desired particles by particle separation means. The predetermined number of particles is dispensed into a receptacle or onto a surface in accordance with a sorting instruction, with the receptacle or surface being positioned by collecting means so as to receive the dispensed particles. A sorting instruction from the particle dispensing means activates the collecting means such that when a sorting instruction has been actuated, so as to deliver a predetermined number of particles into a receptacle or onto a surface which is positioned accurately for sufficient time to collect all sorted particles, the collecting means advances and positions a subsequent surface or receptacle for receipt of particles, the collector means thereafter signalling the particle separation means to commence the next sorting instruction.

Abstract: The present invention makes available a rapid, reproducible, robust assay system for screening and identifying pharmaceutically effective compounds that specifically interact with and modulate the activity of a cellular protein, e.g., a receptor or ion channel. The subject assay enables rapid screening of large numbers of compounds to identify those which act as an agonist or antagonist to the bioactivity of the cellular protein. In particular, the assay of the invention makes use of a cell that harbors a protein that is responsive to a cellular signal transduction pathway. The protein is operatively linked to a polypeptide which causes a detectable signal to be generated upon stimulation of the pathway, e.g., when a compound interacts with and modulates the activity of a cellular receptor or ion channel of the cell.

Abstract: Methods are provided for efficient, high throughput screening of antibody libraries against proteins targets, especially membrane proteins. In particular, methods are provided for screening a fully human antibody library against membrane proteins such as HIV coreceptors in yeast. More particularly, a library of human single chain antibodies is screened against peptide fragments derived from extracellular domains of human CCR5 and high affinity monoclonal antibodies against CCR5 are selected.

Abstract: Methods are provided for generating highly diverse libraries of expression vectors encoding fusion proteins such as single-chain antibodies via homologous recombination in yeast. The method comprises: transforming into yeast cells a linearized yeast expression vector having a 5′- and 3′-terminus sequence at the site of linearization and a library of insert nucleotide sequences that are linear and double-stranded; and having homologous recombination occur between the vector and the insert sequence such that the insert sequence is included in the vector in the transformed yeast cells.

Abstract: Compositions, methods, and kits are provided for efficiently generating and screening a library of highly diverse protein complexes for their ability to bind to other proteins or oligonucleotide sequences. In one aspect of the invention, a library of expression vectors is provided for expressing the library of protein complexes. The library comprises a first nucleotide sequence encoding a first polypeptide subunit; and a second nucleotide sequence encoding a second polypeptide subunit. The first and second nucleotide sequences each independently varies within the library of expression vectors. In addition, the first and second polypeptide subunit are expressed as separate proteins which self-assemble to form a protein complex, such as a double-chain antibody fragment (dcFv or Fab) and a fully assembled antibody, in cells into which the library of expression vectors are introduced.

Abstract: The present invention relates generally to a method of identifying modulators of biological interactions and agents useful for same. More particularly, the present invention contemplates a method of detecting inhibitors of biological interactions involving proteinaceous and/or nucleic acid molecules and more particularly a method of identifying peptide inhibitors of biological interactions having adverse effects on living cells, tissue or organisms. The present invention provides the means by which a wide range of peptide-based therapeutic, prophylactic and diagnostic reagents may be developed.

Abstract: The present invention provides a sensitive test for objectively diagnosing the presence of Didymella bryoniae, the causative agent of gummy stem blight, and differentiating it from similar, nonpathogenic Phoma species. The assay is applicable to DNA isolated from extracts from plant leaves, stem or seed. The detection method employs a polymerase chain reaction technique, using specific oligonucleotide primers for amplification. PCR Products can be visualized using an ELISA-based calorimetric detection system.

Abstract: A yeast cell containing constitutively expressed aequorin and methods of using the cells in growth and toxicity assays are disclosed.

Abstract: A first aspect of the present invention is directed to a transformed yeast cell containing a first heterologous DNA sequence which codes for a G protein-coupled receptor, for example, the somatostatin receptor, and a second heterologous DNA sequence which codes for a G protein a subunit or portions thereof fused to DNA sequences from the yeast G protein in a subunit. A second aspect of the present invention is a transformed yeast cell containing a heterologous DNA sequence which codes for a G protein coupled receptor. A third aspect of the present invention is a method of assaying compounds to determine effects on cell growth.

Abstract: The present invention provides chitin-binding fragments of human chitinase, fragment analogs, purified and isolated polynucleotide sequences encoding such fragments and analogs, and materials and methods for the recombinant production of human chitinase fragment products which are expected to be useful as in products for detecting chitin, binding chitin, and treating fungal infections or for development of products useful for treating the same.

Abstract: A method and apparatus for the differentiation of Crohn's disease from other gastrointestinal illnesses, such as ulcerative colitis and irritable bowel syndrome, using the presence of fecal anti-Saccharomyces cerevisiae antibodies (ASCA) as a marker for Crohn's disease are provided. The apparatus includes an enzyme-linked immunoassay or other immunoassay that utilizes antibodies specific to human immunoglobins for the measurement of total endogenous ASCA in a human fecal sample. The method and apparatus may be used by healthcare providers to distinguish Crohn's disease from other gastrointestinal illnesses, such as ulcerative colitis and irritable bowel syndrome.

Abstract: Disclosed and claimed is the CaESS1 gene, portions thereof such as primers or probes, expression products therefrom, and methods for using the gene, and expression products; for instance, for diagnostic, therapeutic or preventive compositions.

Abstract: The present invention relates to novel yeast cells with increased permeability to compounds, such as small organic compounds. In particular, the invention provides genetically modified yeast cells carrying functional, preferably conditionally regulated, copies of HXT9 and HXT11 genes integrated in the chromosome at the PDR1 and PDR3 loci, thereby disrupting the PDR1 and PDR3 gene activity. The invention further relates to methods and compositions for the use of these hyperpermeable yeast cells for screening for compounds that modulate macromolecular interactions. The invention is exemplified by the use of the hyperpermeable yeast cells in such a screening system. In addition, the invention further provides methods of producing the yeast cells of the invention, as well as polynucleotides, vectors, and kits for use of the hyperpermeable yeast cells and the screening methods of the invention.

Abstract: D-aminoacylase derived from fungi is provided. The fungi capable of producing D-aminoacylase include those belonging to the genus Hypomyces, Fusarium, Auricularia, Pythium, and Menisporopsis. The fungal D-aminoacylase is useful for efficiently producing D-amino acids from N-acetyl-D-amino acids.

Abstract: A kit and method of predicting a refractory response in a subject diagnosed as having periodontal disease by measuring serum concentrations of actinomyces antibodies, streptococcal antibodies and lysine decarboxylase antibodies and using the measurement along with other subject information in a set of derived equations.

Abstract: The invention provides isolated nucleic acids molecules, designated Kinase and Phosphatase nucleic acid molecules, which encode novel protein kinase and protein phosphatase polypeptides. The invention also provides antisense nucleic acid molecules, recombinant expression vectors containing Kinase and Phosphatase nucleic acid molecules, host cells into which the expression vectors have been introduced, and nonhuman transgenic animals in which a Kinase and Phosphatase gene has been introduced or disrupted. The invention still further provides isolated Kinase and Phosphatase proteins, fusion proteins, antigenic peptides and anti-Kinase and Phosphatase antibodies. Diagnostic, screening, and therapeutic methods utilizing compositions of the invention are also provided.

Abstract: A (1→3)-&bgr;-D-glucan binding protein, a fluorescence-labeled (1→3)-&bgr;-D-glucan binding domain protein, a (1→3)-&bgr;-D-glucan measuring agent comprising the same, a method for measuring (1→3)-&bgr;-D-glucan using the same, and a (1→3)-&bgr;-D-glucan assay kit comprising the same.

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

Abstract: A method for determining a cause for digestive and immune disorders is disclosed. The method determines the levels of antibodies against normal intestinal microflora and food antigens. It then compares the results to normal levels to determine the cause. The test can be used to diagnose food allergy or intolerance, microflora imbalance, gut barrier dysfunction, bacterial translocation, immunodeficiencies, candidiasis and autoimmunities.

Abstract: The invention relates to a genes isolated from an oomycete that encode homologs of an FtsZ protein. The invention includes methods of using these proteins to discover new antimicrobials, based on the essentiality of the gene for normal growth and development. The invention can also be used in screening assays to identify inhibitors that are potential antimicrobials. These antimicrobials may be used in a method of controlling oomycete growth on crop plants and seeds.

Abstract: The present invention makes available a rapid, reproducible, robust assay system for screening and identifying pharmaceutically effective compounds that specifically interact with and modulate the activity of a cellular protein, e.g., a receptor or ion channel. The subject assay enables rapid screening of large numbers of compounds to identify those which act as an agonist or antagonist to the bioactivity of the cellular protein. In this system, the first cell is treated with a compound, and functional interaction of this compound with a cellular receptor yields a secreted signal. A second cell, bearing a receptor for this secreted signal, makes use of an indicator gene in a signaling pathway coupled to this second receptor. The subject assays include methods of identifying compounds which specifically modulate, for example, heterologous receptors coupled to the pheromone response pathway in yeast.

Abstract: A method for selecting compounds capable of modulating protein-protein interactions is provided, in which two fusion proteins are prepared and allowed to interact with each other in the presence of test compounds. The interaction between the two fusion proteins leads to protein trans-splicing, producing an active reporter. Compounds that disrupt or enhance protein-protein interactions can be selected based on the presence or absence of the active reporter.

Abstract: Compositions, kits, and methods are provided for use in a recombinational cloning or subcloning methods for constructing expression vectors which comprise: ligating a library of double-stranded linear donor DNAs, where each member of the library includes a donor DNA sequence, with a double-stranded linear driver DNA which includes a promoter sequence and a donor recombination site to form a single circular donor DNA, the single circular donor DNA not including an origin of replication, where the donor DNA sequence is under the transcriptional control of the promoter; and contacting the circular donor DNA and a circular acceptor acceptor vector in the presence of a recombinase to form a single fused circular vector, the circular acceptor vector comprising an origin of replication and an acceptor recombination site capable of recombining with the circular donor DNA.

Abstract: A method and kit for detecting Trichomonas vaginalis infection in a human subject are disclosed. In the method, a body-fluid sample such as a vaginal-swap sample or urine is obtained from the subject and contacted with an antibody specific against a Trichomonas adhesin peptide, forming an antibody-adhesin peptide complex if the subject is infected with Trichomonas. The presence or absence of the complex establishes, with a reliability of at least 80%, in the case of a vaginal swab sample, and with a reliability of at least 40% in the case of a urine sample, the presence or absence presence or absence, respectively, of Trichomonas infection in the subject. A preferred test kit employs a dry-strip, sandwich assay, format.

Abstract: The present invention involves a method and an immuno-analytical device for the rapid and simultaneous detection of multiple micro-organisms in the biological fluids from milk-producing animals suffering from mastitis. This method is based on a lateral flow immuno-assay technique performed to detect antigens specific for multiple infectious agents which are known to cause and/or be encountered in cases of mastitis. Mastitis is an inflammatory condition affecting the udders of milk-producing animals as a result of microbial infections.

Abstract: Screening methods for identifying substances that provide therapeutic value for various diseases associated with protein misfolding are provided. Genetic and chemical screening methods are provided using a yeast system. The methods of the invention provide a rapid and cost-effective method to screen for compounds that prevent protein misfolding and/or protein fibril formation and/or protein aggregation which includes numerous neurodegenerative diseases including Parkinson's disease, Alzheimer's disease, Huntington's disease as well as non-neuronal diseases such as type 2 diabetes.

Abstract: A method to specifically identify transcription factors and transcriptional coactivators from a given cell type and the use of individual clonal cell lines stable expressing gal4-TA domain hybrid proteins for screening or characterization of novel drugs as well as of mixed cell populations stably expressing different gal4-TA domain hybrid proteins for high-throughput screening of novel drugs.

Abstract: The invention relates to a process for screening antimycotically active substances, in which process essential mycete genes, in particular Saccharomyces cerevisae genes and functionally homologous and/or sequentially homologous mycete genes, are employed as targets. A process for finding antimycotically active substances, wherein essential mycete genes are employed as targets.

Abstract: The invention provides novel yeast cells comprising genes whose expression can be modulated by growth in the presence or absence of metal ions, methods for making such yeast cells, and methods of using such yeast cells for determining the requirement for expression of particular genes for the growth or viability of the yeast cells. The invention also provides methods of using such yeast cells in the isolation, screening and analysis of candidate antifungal compounds.

Abstract: This patent describes a novel method of detecting genetic interactions in yeast. This method can also be used to screen for function of biological effectors on yeast. The method encompasses crossing yeast strains with genetic alterations to acquire double mutants. The phenotypes of these double mutants are then checked to detect genetic interactions between the double mutants. This method can be used to assign function to yeast genes and their viral, prokaryotic, and eukaryotic homologs, and aptamers. It can also be used to study yeast two hybrid interactions and to find genes that regulate certain yeast promoters.