Abstract: A culture system for producing PGCs or EG cells by culturing PGCs for long periods in tissue culture is provided. This culture system uses LIF, bFGF, IGF and SCF. The resultant EG cells are useful for the production of transgenic and chimeric avians, in particular, chickens and turkeys, and also for cloning purposes.

Abstract: Transgenic mice that produce high levels of humanized antibodies are described. Targeted gene replacement exchanges constant regions of the mouse immunoglobulin heavy and light chain genes with human genes, either through conventional gene targeting, or by use of the bacteriophage-derived Cre-loxP recombination system. The transgenic animals undergo antibody affinity maturation, and a class switch from the native immunoglobulin to the humanized form.

Abstract: The invention relates to transgenic non-human animals capable of producing heterologous T-cell receptors and transgenic non-human animals having inactivated endogenous T-cell receptor genes. The invention also relates to methods and vectors and transgenes for making such transgenic non-human animals.

Abstract: Vectors and methods are provided for introducing genetic material into cells of a chicken or other avian species. More particularly, vectors and methods are provided for transferring a transgene to an embryonic chicken cell, so as to create a transgenic hen wherein the transgene is expressed in the hen's oviduct and the transgene product is secreted in the hen's eggs and/or those of her offspring. In a preferred embodiment, the transgene product is secreted in the egg white.

Abstract: Proteinaceous products can be produced by transgenic animals having genetic constructs integrated into their genome. The construct comprises a 5′-flanking sequence from a mammalian milk protein gene (such as beta-lactoglobulin) and DNA coding for a heterologous protein other than the milk protein (for example a serin protease such as alpha1-antitrypsin or a blood factor such as Factor VIII or IX). The protein-coding DNA comprises at least one, but not all, of the introns naturally occurring in a gene coding for the heterologous protein. The 5′-flanking sequence is sufficient to drive expression of the heterologous protein.

Abstract: Transgenically produced prolactin and methods of making and using transgenically produced prolactin.

Abstract: The invention described herein allows the production of recombinant retroviruses (retroviral vector particles) from producer cells which are safer and of higher titre than normal. In addition, methods are provided for making helper cells which, when a recombinant retrovirus genome is introduced to make a producer line, produce particles that are targeted toward particular cell types. Methods are also provided for making recombinant retrovirus systems adapted to infect a particular cell type, such as a tumor, by binding the retrovirus or recombinant retrovirus in the particular cell type. Methods are also provided for producing recombinant retroviruses which integrate in a specific small number of places in the host genome, and for producing recombinant retroviruses from transgenic animals.

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: The present invention provides a genetically-modified, non-human mammal having an altered body fat composition, wherein said mammal comprises an exogenous mutant p85 PI3-K gene, wherein the expression of said gene is driven by an insulin responsive cell specific promoter. The present invention also relates to transgenic cell lines containing the same mutant gene, as well as methods using both animals and/or cells.

Abstract: The present invention relates to human bile salt-stimulated lipase (BSSL) obtainable from transgenic sheep. The invention further relates to transgenic sheep whose germ cells and somatic cells contain a recombinant nucleotide molecule comprising a nucleotide sequence encoding for human BSSL. The invention also relates to methods for producing said transgenic animals, as well as to methods for producing human BSSL derived from transgenic animals. In addition, the invention provides the use of compositions comprising BSSL in the treatment of diseases relating to exocrine pancreatic insufficiency, and for improvement of the utilization of dietary lipids in preterm born infants.

Abstract: The intention relates to a method for producing a protein of interest, comprising transforming an insect with a non-viral expression system capable of expressing the protein of interest in the larvae of the insect; breeding the insect to produce larvae; culturing the larvae; and isolating the protein of interest from the larvae. The transformation is advantageously performed by means of a transposon vector.

Abstract: This invention relates to gene delivery and expression, including gene therapy, by using vectors which encode stable mRNA and methods of using such vectors. In particular, this invention relates to vectors which establish controlled expression of recombinant IGF-I genes within tissues at certain levels. The vector includes a 5′ flanking region which includes necessary sequences for expression of a nucleic acid cassette, a 3′ flanking region including a 3′ UTR and/or 3′ NCR, and a linker which connects the 5′ flanking region to a nucleic acid sequence. The linker has a position for inserting a nucleic acid cassette. The linker does not contain the coding sequence of a gene that the linker is naturally associated with. The 3′ flanking region is 3′ to the position for inserting the nucleic acid cassette. The expression vectors of the present invention can also be regulated by a regulatory system and/or constructed with a coating.

Abstract: A method of producing a protein that degrades or detoxifies organic material is described. This method involves producing a non-human transgenic animal that produces such protein in its urine, and has stably integrated into its genome an exogenous gene encoding a protein that is detectable in the urine. Thus, a non-human transgenic animal that produced such protein in its urine, and a method of degrading or detoxifying organic materials also is described. Also a facility comprising a non-human transgenic animal that produce in its urine a protein that degrades or detoxifies organic material and a structure containing such animal is described. A method of altering a substance naturally found in urine is described. A DNA construct used in producing the non-human transgenic animal also is described.

Abstract: In accordance with the present invention, there are provided humanized fish insulin genes. Humanized insulin the present invention encode human insulin alpha and/or beta chains while using fish-preferred codons and regulatory sequences. These humanized genes are thus expressible in fish islet cells. Also provided are transgenic fish having islet cells containing and capable of expressing humanized insulin genes. These islet cells (Brockmann Bodies) can be xenotransplanted into subjects having diabetes. In this manner normoglycemia can be achieved in the recipient of the islets.

Abstract: The present invention provides improved methods for detection of recombinant proteins. The fusion proteins of the invention comprise a capture tag sequence, a detection tag sequence, and polypeptide sequence of interest.

Abstract: There is provided a method for producing a polypeptide using a recombinant baculovirus according to the present invention, wherein said recombinant baculovirus is constructed by ligating the untranslated region of 21 base pairs as set forth in SEQ ID NO:1 upstream of the lobster tropomyosin coding sequence wherein the level of expression of the polypeptide in the baculovirus is remarkably enhanced. There is also provided the polynucleotide sequence of 21 base pairs as set forth in SEQ ID NO:1 and the transfer vector containing the sequence, the recombinant baculovirus genome containing the sequence and the recombinant baculovirus containing the recombinant baculovirus genome to use in the above method.

Abstract: The present invention is directed to DNA constructs suitable for gene expression in mammalian cells and which are characterized by the presence of a mammalian promoter under the control of a tet operator/repressor system. The DNA may be used as part of a system for expressing recombinant protein. In addition, the tet operator/repressor system can be used to engineer cis- and trans-destructive viruses which are capable of replicating in the presence of the tet repressor, but not in the absence of the repressor. These viruses can be used either directly in the treatment of patients with corresponding viral diseases, as vehicles for the delivery of nucleic acids that can serve as therapeutic agents and as part of vaccines designed to immunize people or animals against viral diseases.

Abstract: A microbial adherence inhibitor in the form of fowl egg antibodies is disclosed, along with the method of making it and methods of using it. The inhibitor functions by substantially preventing the attachment or adherence of colony-forming immunogens in the rumen and intestinal tracts of host food animals. The inhibitor is made by inoculating female birds with the immunogen, harvesting the eggs which contain antibodies to the immunogen, harvesting the eggs which contain antibodies to the immunogen, drying the egg contents and adding to the feed or water for the host animals.

Abstract: A microbial adherence inhibitor in the form of fowl egg antibodies is disclosed, along with the method of making it and methods of using it. The inhibitor functions by substantially preventing the attachment or adherence of colony-forming immunogens in the rumen and intestinal tracts of host food animals. The inhibitor is made by inoculating female birds with the immunogen, harvesting the eggs which contain antibodies to the immunogen, harvesting the eggs which contain antibodies to the immunogen, drying the egg contents and adding to the feed or water for the host animals.

Abstract: A method of obtaining a high yield of differentiated human cells and organs includes the steps of providing typed human bone marrow or cord blood stem cells, providing pre-immune non-human mammalian fetuses, implanting the cells into the fetuses, permitting the fetuses to grow for a sufficient time to produce differentiated cells in hybrid organs, and harvesting the differentiated cells from the mammals.

Abstract: A vector is provided which contains a promoter construct linked to a heterologous gene encoding a selected biologically active molecule or oncogene wherein the promoter construct is capable of directing urothelial expression of the heterologous gene. Methods of isolating biologically active molecules from urine of animals transfected with this vector and transgenic animals containing this vector are also provided.

Abstract: A method to prevent graft rejection of transplanted cells, tissues or organs without general immunosuppression is described. The method employs a newly discovered protein, LAG-3. When allogeneic or xenogeneic cells are engineered to express LAG-3 on their surface and transplanted, immune destruction of the implanted cell, tissue or organ is prevented, while the host's immune system remains functional. A particular application of this method allows the preparation of a universal gene therapy host cell expressing LAG-3 on its surface for protection from graft rejection by a host's immune system.

Abstract: The present invention is directed toward a method to produce recombinant fusion proteins in large quantities that are both highly homogenous and biologically active. In particular, the invention relates to a method for producing recombinant fusion proteins in a larvae expression system. The recombinant fusion protein is then purified from the larvae by an affinity tag fused to the protein via affinity chromatography.

Abstract: Switch regions derived from an immunoglobulin (Ig) gene are used to direct recombination between a targeting construct containing a promoter, a switch region (S1), and 2) a target locus minimally containing a promoter, a switch region (S2), and a target sequence.

Abstract: A transgenic animal with alterations in an H2-O gene is prepared by introduction of an altered H2-O gene into a host animal. The resulting transgenic animals produce a substantially greater frequency of high affinity antibodies compared to H2-O wild type animals. A method for the production of high affinity antibodies is disclosed.

Abstract: A method of fetal gene therapy is disclosed. In general, the method comprises the steps of identifying a fetus with a genetic defect, obtaining allantois/umbilical cord cells expressing a gene product that ameliorates the genetic defect, and exposing the fetus to the allantois/umbilical cord cells wherein a chimeric allantois is capable of supplying the gene product to the fetus is created. The present invention is also a method of examining the effect of test compounds on vasculogenesis and angiogenesis by observing the effect of the test compound on cultured allantoic explants.

Abstract: The present invention relates to a method of producing osteocyte cell line in various stages of differentiation. Such cell line remains stable after more than 20 passages. The osteocyte has a stellate shape with dendritic processes and expresses high level of osteocalcin. More specifically, it provides a method of production for cultured osteocytes of various differentiation stages. Furthermore, it relates to osteocyte cell line, and more specifically cultured osteocyte. The invention also relates to a method for the production of monoclonal antibodies using such cultured osteocytes and further relates to hybridomas and monoclonal antibodies which recognize an osteocyte-specific antigen. Finally, the invention relates to a method of screening for modification factors and binding factors for osteocytes.

Abstract: The present invention provides a method of biasing the immune response of a mammal toward a desired epitope of a chosen antigen, particularly a functionally-relevant epitope. In preferred embodiments, the epitope-biasing method leads to fully-human antibodies of defined specificity with affinities of 10 nM to 50 pM. The invention further provides antibody libraries biased to tissues and to cell types, for use in generating epitope expression profiles useful for characterizing unknown genes. When all aspects of the present invention are combined, they result in an integrated system for defining critical epitopes on newly discovered gene products and rapidly devloping therapeutic grade antibodies to those critical epitopes.

Abstract: Described is a method of targeting specific genes to the mammary gland which results in the efficient synthesis and secretion of biologically important molecules. Further, there is described as a composition of matter, a transgenic mammal having the ability to reproduce itself and being suitable for the secretion of biologically active agents into its milk. Additionally there is disclosed as a composition of matter, recombinant DNA gene complexes designed to integrate into a mammalian genome and to synthesize and secrete biological active agents into the milk. Furthermore methods of producing and using altered milk are disclosed.

Abstract: The invention features a transgenic animal, which expresses a Coxsackie-adenovirus receptor (CAR) in its cells. The invention provides a new animal model for effecting adenovirus-mediated gene delivery and for testing gene function in vivo, and also provides new opportunities to assess gene function in vitro using freshly isolated cells.

Abstract: This invention is a transgenic fish that expresses an amino acid sequence (either peptide or protein) under control of a chemical substance when the chemical substance is supplied to the fish. The protein will preferably be a heterologous protein, such as a protein useful as a pharmaceutical product in humans, or animals. The chemical substance may be a hormone or hormone mimic, such as a steroid, thyroid, retinoid and vitamin D. Especially preferred are fish responsive to estrogens and having estrogen responsive elements in the regulatory sequences for a heterologous protein. The transgenic fish may express a desired heterologous protein in a specific tissue such as a particular organ, especially preferred fish expresses a heterologous protein or peptide in the liver. Another preferred fish expresses a protein or peptide in the egg.

Abstract: A vector is provided which contains a promoter construct linked to a heterologous gene encoding a selected biologically active molecule or oncogene wherein the promoter construct is capable of directing urothelial expression of the heterologous gene. Methods of isolating biologically active molecules from urine of animals transfected with this vector and transgenic animals containing this vector are also provided.

Abstract: A method and system for controlling the expression of transgene products in specific tissues in a transgenic animal is provided. The method comprises: a) providing a fist transgenic parent animal whose genome comprises an F transgene comprising an exogenous gene operatively linked to a promoter that is upregulated by a transactivator protein; b) providing a second transgenic parent animal whose genome comprises i) a second transgene, comprising a second promoter that is downregulated by the transactivator protein and operatively linked to an antisense gene that encodes a sequence which inhibits or reduces processing of the F transgene transcript; and ii) a third transgene comprising a tissue specific promoter operatively linked to a gene encoding the transactivator protein; and c) breeding the first transgenic parent animal with the second transgenic parent animal to provide a transgenic offspring animal whose genome comprises the three transgenes.

Abstract: Variant human MLH1 and MSH2 genes are provided. Methods of using these variant genes to diagnose hereditary non-polyposis colorectal cancer (HNPCC) and/or determine a patient's susceptibility to developing HNPCC are also provided. Methods and compositions for identifying new variant MLH1 of MSH2 genes are also provided. In addition, experimental models for hereditary non-polyposis colorectal cancer comprising these variant genes are provided.

Abstract: Disclosed is a method for the recombinant production of biofilaments, such as spider silk or insect fibroins, using transgenic animals which secrete the biofilaments in their milk and/or urine, and transgenic cells which secrete the biofilaments into culture media. Such a method is useful for producing large quantities of biofilament material. Also disclosed is a nucleic acid molecule for generating such transgenic animals.

Abstract: The present invention describes a non-human transgenic mouse that produces in its leukocytes, a recombinant human leukotriene B4 receptor (BLTR), having physiological activity of human BLTR. The transgenic mouse has stably integrated into its genome an exogenous gene construct which includes (A) 5′ expression regulating sequences, including a BLTR specific promoter, (B) DNA encoding the BLTR and a signal sequence effective in directing overexpression of the BLTR into leukocytes of the transgenic mouse and (C) 3′ regulatory sequences that result in the overexpression of the DNA in the leukocytes. In one embodiment, (A), (B), and (C) are operably linked in the gene construct to obtain production of the BLTR in the leukocytes and overexpression thereof in the transgenic mouse.

Abstract: Methods are described to obtain nucleic acid molecules that encode T cell receptors and their derivatives that are human HLA-restricted and which are specific for tumor-associated antigens found in human tumors. These nucleic acids are useful in preparing recombinant cells for diagnosis and therapy of human tumors.

Abstract: The subject invention relates to methods of producing non-human transgenic mammals which produce various oligosaccharides and glycoconjugates in their milk. Additionally, the subject invention relates to the mammals themselves, the milk which they produce, compositions comprising the milk, fractions of the milk, and the purified oligosaccharides, as well as glyconjugates, present in the milk.

Abstract: The present invention relates to a method for the production and secretion into animal's semen of an exogenous recombinant protein comprising the steps of: a) producing a non-human transgenic animal characterized by an expression system comprising a promoter specific for the genital tract or accessory glands operatively linked to an exogenous DNA sequence coding for the recombinant protein through a DNA sequence coding for a signal peptide effective in secreting and maturing the recombinant protein in genital tract tissue; b) collecting semen produced by the non-human transgenic animal; and c) isolating the exogenous recombinant protein from the semen.

Abstract: Peptides can be produced in and purified from the milk of transgenic animals. The peptides are made as fusion proteins with a suitable fusion partner such as &agr;-lactalbumin, which is a natural milk protein. The fusion partner protein acts to promote secretion of the peptides and, at least in the case of &agr;-lactalbumin, allows a single-step purification based on specific affinity. The peptide is released from the purified fusion protein by a simple cleavage step and purified away from the now liberated &agr;-lactalbumin by repeating the same affinity purification method. A particular advantage of producing peptides via this route, in addition to the obvious advantages of high yield and biocompatibility, is that specific post-translational modifications, such as carboxy terminal amidation, can be performed in the mammary gland.

Abstract: The subject invention provides non-human mammalian hosts characterized by inactivated endogenous Ig loci and functional human Ig loci for response to an immunogen to produce human antibodies or analogs thereof. The hosts are produced by multiple genetic modifications of embryonic cells in conjunction with breeding. Different strategies are employed for recombination of the human loci randomly or at analogous host loci. Chimeric and transgenic mammals, particularly mice, are provided, having stably integrated large, xenogeneic DNA segments. The segments are introduced by fusion with yeast spheroplasts comprising yeast artificial chromosomes (YACs) which include the xenogeneic DNA segments and a selective marker such as HPRT, and embryonic stem cells.

Abstract: DNA constructs are provided of epitope-tagged proteins or protein fragments which are conveniently purified with immunoaffinity chromatography such as epitope-tagged prion proteins (PrP). Transgenic animals expressing an epitope-tagged protein are provided, including transgenic animals expressing epitope-tagged PrP. Methods for distinguishing between the conformational shapes of a protein and a convenient method for isolating a tagged protein by immunoaffinity chromatographic methods are provided.

Abstract: Transgenes for producing recombinant polypeptides transgenic bovine species. A transgene for producing recombinant polypeptides in the milk of transgenic bovine species comprises at least one expression regulation sequence, a secretory DNA sequence encoding a secretory signal sequence which is functional in mammary secretory cells of the bovine species and a recombinant DNA sequence encoding the recombinant polypeptide. Also included are methods for producing transgenic bovine species. The method includes introducing the above transgene into an embryonal target cell of a bovine species, transplanting the transgenic embryonic target cell formed thereby into a recipient bovine parent and identifying at least one female offspring which is capable of producing the recombinant polypeptide in its milk.

Abstract: A method to produce a cell expressing an antibody from a genomic sequence of the cell comprising a modified immunoglobulin locus using Cre-mediated site-specific recombination is disclosed. The method involves first transfecting an antibody-producing cell with a homology-targeting vector comprising a lox site and a targeting sequence homologous to a first DNA sequence adjacent to the region of the immunoglobulin loci of the genomic sequence which is to be converted to a modified region, so the first lox site is inserted into the genomic sequence via site-specific homologous recombination. Then the cell is transfected with a lox-targeting vector comprising a second lox site suitable for Cre-mediated recombination with the integrated lox site and a modifying sequence to convert the region of the immunoglobulin loci to the modified region.

Abstract: The subject invention provides non-human mammalian hosts characterized by inactivated endogenous Ig loci and functional human Ig loci for response to an immunogen to produce human antibodies or analogs thereof. The hosts are produced by multiple genetic modifications of embryonic cells in conjunction with breeding. Different strategies are employed for recombination of the human loci randomly or at analogous host loci. Chimeric and transgenic mammals, particularly mice, are provided, having stably integrated large, xenogeneic DNA segments. The segments are introduced by fusion with yeast spheroplasts comprising yeast artificial chromosomes (YACs) which include the xenogeneic DNA segments and a selective marker such as HPRT, and embryonic stem cells.

Abstract: A method to produce a cell expressing an antibody from a genomic sequence of the cell comprising a modified immunoglobulin locus using Cre-mediated site-specific recombination is disclosed. The method involves first transfecting an antibody-producing cell with a homology-targeting vector comprising a lox site and a targeting sequence homologous to a first DNA sequence adjacent to the region of the immunoglobulin loci of the genomic sequence which is to be converted to a modified region, so the first lox site is inserted into the genomic sequence via site-specific homologous recombination. Then the cell is transfected with a lox-targeting vector comprising a second lox site suitable for Cre-mediated recombination with the integrated lox site and a modifying sequence to convert the region of the immunoglobulin loci to the modified region.

Abstract: Antibodies with fully human variable regions against a specific antigen can be prepared by administering the antigen to a transgenic animal which has been modified to produce such antibodies in response to antigenic challenge, but whose endogenous loci have been disabled. Various subsequent manipulations can be performed to obtain either antibodies per se or analogs thereof.

Abstract: A monoclonal preparation of lymphocytes, their production and use.

Abstract: The invention relates to a method for making monoclonal antibodies having pre-defined specificity for an epitope characteristic of or unique to a single form of a polymorphic protein. The method includes constructing a first transgenic animal to express a first form of a polymorphic protein encoded by a first allele of a gene encoding the protein; constructing a second transgenic animal to express a second form of the polymorphic protein encoded by a second allele of the gene encoding the protein; and immunizing the first transgenic animal with cells from the second transgenic animal expressing the second form of the polymorphic protein to induce an immune response in the first transgenic animal yielding an antibody specific for an epitope peculiar to the second form of the polymorphic protein. The invention further includes hybridoma cells secreting a monoclonal antibody specific for the second form of the protein.

Abstract: The present invention relates to a transgenic non-human mammal comprising a DNA sequence encoding human extracellular superoxide dismutase (human EC-SOD) or a variant thereof which is expressed in the milk. Transgenic mice containing a chimeric whey acidic protein gene promoter operatively linked to human EC-SOD gene were produced. Levels of up to 0.7 mg human EC-SOD protein/mL milk were observed. The mammalian expression system is preferably expressed in a non-human mammal selected from the group containing rabbits, mice, rats, goats, sheep, pigs, llama, camels and bovine species. The human EC-SOD proteins dismutate superoxide radicals and bind heparin. Within the scope of the invention are also method for producing a transgenic non-human mammal capable of expressing human EC-SOD as defined above, and methods of making milk and methods of isolating protein from the milk.