Abstract: The invention relates to a process for the genetic selection in microorganisms of proteins which are capable of ligand binding, in which process a protein which is capable of ligand binding is presented extracytoplasmically and the signal of the ligand binding is passed on by signal transduction to the biosynthetic machinery of the micoorganism for the purpose of expressing a detectable and/or selectable function. In addition to this, the patent discloses microorganisms which are suitable for use in this process, as well as replicons and processes for their preparation.

Abstract: A method of overproducing proteins which are initiated with an amino acid other than methionine, in which methionyl-tRNA transformylase and, optionally, the appropriate tRNA synthetase are overexpressed in a host cell.

Abstract: A non-human transgenic mammalian animal, as described above, contains an exogenous double stranded DNA sequence stably integrated into the genome of the animal, which comprises cis-acting regulatory units operably linked to a DNA sequence encoding human Factor VIII protein and a signal peptide, where the cis-acting regulatory units are active in mammary gland cells and the signal peptide is active in directing newly expressed Factor VIII into the milk of the animal. The promoter may be a milk protein promoter such as for whey acidic protein, casein, lactalbumin, or beta-lactoglobulin promoter. The transgenic mammals are preferably farm animals, for example, cows, goats, sheep, rabbits and pigs. Concurrent expression of a gene for human von Willebrand's Factor into milk may be used to stabilize newly-secreted Factor VIII.

Abstract: The present invention relates to an improved system for producing mature heterologous proteins, and especially hirudin, by means of yeasts comprising an expression vector containing a sequence coding for the heterologous protein. This improvement is characterized by amplification of the KEX2 gene of yeast, coding for the endoprotease yscF. The amplification is carried out either by integration of one or more copies of all or part of the KEX2 gene in the yeast genome, or by insertion of one or more copies of all or part of the KEX2 gene into the vector for expression of the heterologous protein.

Abstract: A method for introducing and expressing genes in animal cells is disclosed comprising infecting said animal cells with live invasive bacteria, wherein said bacteria contain a eukaryotic expression cassette encoding said gene. The gene may encode, e.g., a vaccine antigen, an therapeutic agent, an immunoregulatory agent or a anti-sense RNA or a catalytic RNA.

Abstract: The present invention discloses a selection marker free system which can be used to introduce genetic modifications in bacteria, yeasts and fungi. The system can be employed to introduce or delete desired genes or DNA fragments in the genome of the indicated host species without leaving any undesired DNA i.e. the selection marker used for selection of transformants or other DNA used for cloning. In this way strains have been developed containing only desired genes introduced at desired chromosomal sites. Similarly, desired DNA fragments have been deleted or replaced at desired sites.

Abstract: Methods of inducing tolerance including administering to the recipient a short course of help reducing treatment or administering a short course and methods of prolonging the acceptance of a graft by administering a short course of an immunosuppressant.

Abstract: The invention relates to transgenic non-human animals capable of producing heterologous antibodies, i.e., antibodies encoded by immunoglobulin heavy and light chain genes not normally found in the genome of that species of non-human animal. In one aspect of the invention, transgenes encoding unrearranged heterologous human immunoglobulin heavy and light chains are introduced into a non-human animal thereby forming a transgenic animal capable of producing antibodies encoded by human immunoglobulin genes. Such heterologous human antibodies are produced in B-cells which are thereafter immortalized, e.g., by fusing with an immortalizing cell line such as a myeloma or by manipulating such B-cells by other techniques to perpetuate a cell line capable of producing a monoclonal heterologous antibody.

Abstract: A method of preventing tumor cell metastasis by inhibiting the binding of hepatocyte growth factor/scatter factor ("HGF/SF") with met proto-oncogene protein is described. A method of producing HGF/SF and a cell line for the production of HGF/SF are also described. The met proto-oncogene tyrosine kinase receptor (Met) and its ligand, hepatocyte growth factor/scatter factor (HGF/SF), ordinarily constitute a paracrine signalling system in which cells of mesenchymal origin produce the ligand, which binds to the receptor that is predominantly expressed in cells of epithelial origin. The method of the present invention disrupts the Met-HGF/SF autocrine signaling that contributes to the tumorigenic process in tumors of mesenchymal origin, such as sarcomas.

Abstract: A method of reducing an immune response to a recombinant adenovirus which involves co-administration of the recombinant adenovirus and a selected immune modulator. The immune modulator functions by inhibiting the formation of neutralizing antibodies and/or reducing CTL killing of the virally infected cells. The method additionally encompasses the step of re-administering the recombinant adenovirus.

Abstract: The striking compartmentalization of p21.sup.WAF1 expression found in normal tissues is completely abrogated in neoplastic tissues. Methods are provided for using p21.sup.WAF1 expression as a tool to assess neoplasia and to discover new drugs. A truncated p21.sup.WAF1 protein is more active than full-length p21.sup.WAF1 protein in inhibiting tumor cell growth.

Abstract: The present invention provides for in vivo gene transfer to adipocytes mediated by adenovirus and, in particular, the in vivo transfer of toxic genes as a means of reducing adiposity, as well as the transfer of genes encoding angiogenic substances to induce new blood vessel growth. The present invention also provides for the in vivo gene transfer to adipocytes to supply a source of proteins to be used in the local milieu of the adipocyte tissue or to be secreted and used systemically. Further, the present invention provides for the transfer of the adipocytes to other sites within a host, following adenoviral-mediated transfer of genes to the adipocytes in vivo, to allow for the exploitation of the modified adipocytes as a transferable means for the production of protein.

Abstract: The present invention is based upon the observation that the temperature sensitive phenotype of the cp45 strain of HPIV-3 correlates to a mutation in the large, or L, gene of cp45 relative to the corresponding gene in the wild-type strain. This correlation enables new vaccines directed at viruses other than HPIV-3 by combining, through genetic engineering methods, the region of the cp45 viral genome which encodes proteins responsible for replication and internal structure with the region of the genome of the target virus which encodes proteins responsible for attachment, penetration and release of the virus and virus progeny, respectfully. Moreover, it is possible to determine whether HPIV-3 or a cp45-hybrid virus is attenuated by confirming the presence or absence of mutations in its L gene.

Abstract: A new technique for generating mixtures of oligonucleotides in a single automated synthesis is taught. The method can be used to prepare mixed oligonucleotides ideally suited for creation of useful mixtures of oligo- or polypeptides or proteins. Additionally, the technique enables insertion and/or substitution and/or deletion of a nucleotide sequence at one or more sites. For protein mutagenesis, a trinucleotide can be inserted or substituted at codon boundaries. The invented technique makes possible the encoding of all possible single amino acid insertions, or any desired mixture of substitutions and insertions.

Abstract: Materials and methods for homologous-recombination screening of DNA libraries constructed in a eukaryotic host and methods for homologous-recombination chromosome walking for isolating overlapping DNA sequences for building an extended physical map of a chromosomal region.

Abstract: Transgenic non-human mammals and methods of preparing such mammals are disclosed. Homologous recombination is employed to inactivate genes, particularly genes coding for neuronal intermediate filaments. These animals, whose phenotype is characterized by reduced or eliminated levels of a neuronal intermediate filament, are useful as models for studying aging, injury to the nervous system, and pathogenesis of neurodegenerative diseases. They are also useful for the screening of potential therapeutic agents.

Abstract: The present invention concerns defective recombinant adenoviruses containing an inserted gene encoding apolipoproteins, pharmaceutical compositions comprising the adenovirus, and their use for the treatment or prevention of pathologies linked to dyslipoproteinemias.

Abstract: Transgenic animals carrying a transgene comprising a nucleic acid molecule encoding protein useful for regulating the expression of genes in eukaryotic cells and organisms in a highly controlled manner are disclosed. In the regulatory system of the invention, transcription of a tet operator-linked nucleotide sequence is inhibited by a transcriptional inhibitor fusion protein composed of two polypeptides, a first polypeptide which binds to tet operator sequences and a second polypeptide which directly or indirectly inhibits transcription in eukaryotic cells. In various embodiment, the first polypeptide binds to tet operator sequences either: (i) in the absence but not the presence of tetracycline (or an analogue thereof) or (ii) in the presence but not the absence of tetracycline (or an analogue thereof). In a preferred embodiment, the transgene encoding the transcriptional inhibitor fusion protein is integrated at a predetermined location within the chromosome of the transgenic animal.

Abstract: Disclosed are methods for accurately determining the total emission intensity of a single fluorochrome, under imaging conditions, using a digital imaging fluorescence microscopy system. Also disclosed are methods for detecting and localizing probe-target molecule binding. The detection method has sufficient resolution and sensitivity to locate and detect a single target-bound probe.

Abstract: The invention provides methods and compositions for evaluating modulators of the Stat6 signaling pathway; in a particular, transgenic mice comprising a transgene within a Stat6 allele locus, encoding a selectable marker and displacing the SH2-encoding domain of the Stat6 allele. More particularly, the transgene may comprise 3' and 5' regions with sufficient complementarity to the natural Stat6 allele at the locus to effect homologous recombination of the transgene with the Stat6 allele. Such mice provide useful animal models for determining the effect of candidate drugs on a host deficient in Stat6 function. The invention provides a variety of methods for making and using the subject compositions; in particular, methods for determining the effect of a candidate drug on a mouse deficient in Stat6 function and methods of evaluating the side effects of a Stat6 function inhibitor.