Abstract: Disclosed herein are compositions and methods relating to peptides. The peptides can inhibit amyloid beta (A?) generation and reduce GSK-3 activities. Further provided are compositions and methods for treating or preventing, for example, Alzheimer's disease, cancer, and diabetes.

Abstract: The present invention provides methods for identifying peptides in a mammalian. Tsg101 protein that binds to the PTAPP (SEQ ID NO: 3) motif or L domain of human immunodeficiency virus type I (HIV-1). Such peptides can be used to inhibit Tsg101-HIV Gag binding, and is therefore effective in reducing HIV particle production. The invention also provides the peptides identified by the method of the invention and to method of using such peptides for treating HIV infection.

Abstract: Methods and compositions for rapidly identifying CGEPs required for viral infection of mammalian cells are provided. Also provided are methods of inhibiting viral infection of mammalian cells by inhibiting the activity of one or more CGEPs (e.g., as identified in accordance with methods of the invention) in the cells. Aspects of the invention further include specifically identified CGEPs implicated in mammalian cell infection of specific viruses, e.g., African Swine Fever Virus and Foot and Mouth Virus, and methods of modulating their activity to achieve viral resistance.

Abstract: TSG101 is a tumor susceptibility gene whose homozygous functional knock out in fibroblasts leads to transformation and the ability of these cells to form metastatic tumors in nude mice. The cellular transformation that results from inactivation of TSG101 is reversible by restoration of TSG101 function. Decreased expression of TSG101 is associated with the occurrence of certain human cancers, including breast carcinomas. The TSG101 nucleic acid compositions find use in identifying homologous or related proteins and the DNA sequences encoding such proteins; in producing compositions that modulate the expression or function of the protein; and in studying associated physiological pathways. In addition, modulation of the gene activity in vivo is used for prophylactic and therapeutic purposes, such as treatment of cancer, identification of cell type based on expression, and the like.

Abstract: Poly G tails prolong mRNA chemical and functional half life in E. coli cell extracts and dramatically increased RNA-dependent protein synthesis in vitro. The effect of polyguanylation on mRNA functional half life, as measured by the ability of CAT transcripts to produce biochemically-active protein in vitro, was four- to six-fold greater than the effect on chemical half life. Addition of a few nucleotides 5′ to the bacteriophage T7 promoter markedly enhanced transcription of linear PCR-generated DNA molecules by T7 RNA polymerase. Collectively a novel approach is provided for efficient in vitro protein synthesis that bypasses the need for cloned DNA templates to obtain the products of translational open reading frames.

Abstract: The present invention provides methods and compositions for regulating ubiquitination in a cell. In particular, the present invention provides purified polypeptides comprising an ubiquitination-regulating domain. The invention also provides methods of using such polypeptides for screening for agents, for producing antibodies, and for treatment of diseases, e.g., proliferative diseases, neurodegenerative diseases, autoimmune diseases, metabolic disease and developmental abnormalities. The invention further provides antibodies that bind an ubiquitination-regulating domain and agents and antibodies that regulate ubiquitination in cells, e.g., by modulating the interaction between a TSG101 protein and an MDM2 protein.

Abstract: TSG101 is a tumor susceptibility gene whose homozygous functional knock out in fibroblasts leads to transformation and the ability of these cells to form metastatic tumors in nude mice. The cellular transformation that results from inactivation of TSG101 is reversible by restoration of TSG101 function. Decreased expression of TSG101 is associated with the occurrence of certain human cancers, including breast carcinomas. The TSG101 nucleic acid compositions find use in identifying homologous or related proteins and the DNA sequences encoding such proteins; in producing compositions that modulate the expression or function of the protein; and in studying associated physiological pathways. In addition, modulation of the gene activity in vivo is used for prophylactic and therapeutic purposes, such as treatment of cancer, identification of cell type based on expression, and the like.

Abstract: TSG101 is a tumor susceptibility gene whose homozygous functional knock out in fibroblasts leads to transformation and the ability of these cells to form metastatic tumors in nude mice. The cellular transformation that results from inactivation of TSG101 is reversible by restoration of TSG101 function. Decreased expression of TSG101 is associated with the occurrence of certain human cancers, including breast carcinomas. The TSG101 nucleic acid compositions find use in identifying homologous or related proteins and the DNA sequences encoding such proteins; in producing compositions that modulate the expression or function of the protein; and in studying associated physiological pathways. In addition, modulation of the gene activity in vivo is used for prophylactic and therapeutic purposes, such as treatment of cancer, identification of cell type based on expression, and the like.

Abstract: TSG101 is a tumor susceptibility gene whose homozygous functional knock out in fibroblasts leads to transformation and the ability of these cells to form metastatic tumors in nude mice. The cellular transformation that results from inactivation of TSG101 is reversible by restoration of TSG101 function. Decreased expression of TSG101 is associated with the occurrence of certain human cancers, including breast carcinomas. The TSG101 nucleic acid compositions find use in identifying homologous or related proteins and the DNA sequences encoding such proteins; in producing compositions that modulate the expression or function of the protein; and in studying associated physiological pathways. In addition, modulation of the gene activity in vivo is used for prophylactic and therapeutic purposes, such as treatment of cancer, identification of cell type based on expression, and the like.

Abstract: TSG101 is a tumor susceptibility gene whose homozygous functional knock out in fibroblasts leads to transformation and the ability of these cells to form metastatic tumors in nude mice. The cellular transformation that results from inactivation of TSG101 is reversible by restoration of TSG101 function. Decreased expression of TSG101 is associated with the occurrence of certain human cancers, including breast carcinomas. The TSG101 nucleic acid compositions find use in identifying homologous or related proteins and the DNA sequences encoding such proteins; in producing compositions that modulate the expression or function of the protein; and in studying associated physiological pathways. In addition, modulation of the gene activity in vivo is used for prophylactic and therapeutic purposes, such as treatment of cancer, identification of cell type based on expression, and the like.

Abstract: Methods are provided for identifying a gene at a random chromosomal locus in the genome of a mammalian cell. The method involves inactivating one copy of the gene by integrating one DNA construct (knockout construct) in that gene copy. The knockout construct includes a positive selection marker region sequence and, in a 5' direction from the selection marker region sequence, a transcription initiation region sequence responsive to a transactivation factor, said transcription initiation region oriented for antisense RNA transcription in the direction away from the selection marker region sequence. The second copy of the gene is inactivated by transforming the cells with a second DNA construct (transactivation construct) containing a gene sequence for the transactivation factor which initiates antisense RNA transcription extending from the knockout construct into the chromosomal locus flanking the knockout construct at its 5' end.

Abstract: Expression systems are provided, where plasmids can be safely maintained in a prokaryotic host, by providing for a tRNA gene for an essential tRNA in a tRNA gene negative background. A non-selective medium can be employed to permit vigorous growth of the host and efficient expression of a protein of interest.

Abstract: Methods and compositions are provided for producing heterologous proteins in a micrcorganism host, particularly bacterium, without fusion to an endogenous protein. Particularly, the heterologous gene may be inserted into a vector comprising an endogenous gene, where the heterologous gene is preceded by a ribosomal binding site. A heterologous enzyme functional in a bacterial host is demonstrated.

Abstract: Method and compositions are provided for replication and expression of exogenous genes in microorganisms. Plasmids or virus DNA are cleaved to provide linear DNA having ligatable termini, which are bound to a gene having complementary termini, to provide a biologically functional replicon with a desired phenotypical property. The replicon is inserted into a microorganism cell by transformation. Isolation of the transformants provides cells for replication and expression of the DNA molecules present in the modified plasmid. The method provides a convenient and efficient way to introduce genetic capability into microorganisms for the production of nucleic acids and proteins, such as medically or commercially useful enzymes, which may have direct usefulness, or may find expression in the production of drugs, such as hormones, antibiotics, or the like, fixation of nitrogen, fermentation, utilization of specific feedstocks, or the like.

Abstract: Method for preparing peptides free of undesired amino acids or amino acid sequences employing site specific receptors and proteases to cleave unprotected enzymatically hydrolyzable bonds.

Abstract: Monoclonal antibodies having conformation-dependent specificity for native dsDNA, as exemplified by the IgM antibody produced by murine hybridoma ATCC No HB 8329. These antibodies are used to detect DNA duplex formation in DNA hybridization tests.

Abstract: Method for preparing high signal strength promoters and terminators and DNA compositions employing such promoters and terminators. T5 phage is cleaved to provide for DNA sequences having intact promoters. These promoters are inserted into vectors separated from a balanced terminator by a gene of interest and the terminator is desirably followed by a marker allowing for selection of transformants. High efficiencies in transcription of DNA can be achieved with the highly active T5 promoters. The promoters and terminators are used in hybrid DNA for efficient expression of structural genes and transcription to provide RNA sequences.

Abstract: Method and compositions are provided for replication and expression of exogenous genes in microorganisms. Plasmids or virus DNA are cleaved to provide linear DNA having ligatable termini, which are bound to a gene having complementary termini, to provide a biologically functional replicon with a desired phenotypical property. The replicon is inserted into a microorganism cell by transformation. Isolation of the transformants provides cells for replication and expression of the DNA molecules present in the modified plasmid. The method provides a convenient and efficient way to introduce genetic capability into microorganisms for the production of nucleic acids are proteins, such as medically or commercially useful enzymes, which may have direct usefulness, or may find expression in the production of drugs, such as hormones, antibiotics, or the like, fixation of nitrogen, fermentation, utilization of specific feedstocks, or the like.

Abstract: Hollow fiber reactors for growing microbial cells. Isotropic hollow fibers are supported in a housing inoculated with cells. Nutrient medium passing through the lumen undergoes a pressure drop resulting in radial convective flow: the nutrient medium flows outwardly from the lumen into the surrounding area adjacent the entry port and fluid surrounding the hollow fiber flows into the lumen adjacent the exit port. With the efficient distribution of nutrients and removal of product, high cell densities are achieved providing for high product yields per unit reactor volume.

Abstract: Methods for microbiological processing of organic materials for production of valuable products. Asymmetric hollow fibers are employed in a flow reactor, where the hollow fibers have a semipermeable membrane surrounding a lumen, where the semipermeable membrane is supported by a sponge structure. The pores of the sponge structure serve as a housing for microorganisms or cells with high density packing of the microorganisms or cells in the pores. Nutrient medium continuously flowing through the lumen provides nutrients to the microorganisms or cells as well as any substrates to be processed by the microorganisms or cells. The nutrients and substrates diffuse through the semipermeable membrane into the pores, where they are processed, and the metabolic products diffuse into the lumen. The lumen effluent is then processed for the desired products. Optionally, oxygen is provided external to the hollow fiber to enhance the amount of oxygen available to the microorganisms and cells.