Yeast Is A Host For The Plasmid Or Episome Patents (Class 435/483)
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Patent number: 6342373Abstract: The invention relates to DNA sequences which code an eglin, expression plasmids containing such DNA sequences, hosts transformed with such expression plasmids, novel eglin compounds produced from such transformed hosts, monoclonal antibodies against eglins, hybridoma cells which produce such antibodies, and test kits for immunoassays containing such antibodies, and furthermore the processes for their preparation and a process for the preparation of eglins with the aid of the transformed hosts. The eglins which can be prepared according to the invention have useful pharmacological properties.Type: GrantFiled: May 24, 1991Date of Patent: January 29, 2002Assignee: UCP GEN-Pharma AGInventors: Hans Rink, Manfred Liersch, Peter Sieber, Werner Rittel, Francois Meyer, Ursula Seemüller, Hans Fritz, Walter Märki, Sefik Alkan
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Publication number: 20020004243Abstract: A method of introducing a property of a particular yeast strain, based on a recessive allele, into the genetic background of an industrial baker's yeast; as well as yeast strains obtainable according to the method. In particular, a method is disclosed to introduce an lti-property into the genetic background of industrial baker's yeast. The novel strains obtained according to the method may be used for the preparation of a dough and for the manufacture of baked products therefrom, such as on an industrial scale.Type: ApplicationFiled: April 18, 2001Publication date: January 10, 2002Inventors: Christof Gysler, Peter Niederberger
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Publication number: 20020004197Abstract: Methods and nucleic acid constructs for testing and validating activity reporter cells, using a plurality of host cell genes, are described.Type: ApplicationFiled: August 28, 2001Publication date: January 10, 2002Applicant: Iconix Pharmaceuticals, Inc.Inventors: Teri Melese, Edward L. Perkins, Stuart Tugendreich
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Patent number: 6331415Abstract: The invention relates to processes for producing an immunoglobulin or an immunologically functional immunoglobulin fragment containing at least the variable domains of the immunoglobulin heavy and light chains. The processes can use one or more vectors which produce both the heavy and light chains or fragments thereof in a single cell. The invention also relates to the vectors used to produce the immunoglobulin or fragment, and to cells transformed with the vectors.Type: GrantFiled: June 10, 1988Date of Patent: December 18, 2001Assignee: Genentech, Inc.Inventors: Shmuel Cabilly, Herbert L. Heyneker, William E. Holmes, Arthur D. Riggs, Ronald B. Wetzel
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Patent number: 6329141Abstract: The present invention provides recombinant DNA comprising a transcription promoter and a downstream sequence to be expressed, in operable linkage therewith, wherein the transcription promoter comprises a region found upstream of the open reading frame of a highly expressed Phaffia gene, preferably a glycolytic pathway gene, more preferably the gene coding for Glyceraldehyde-3-Phosphate Dehydrogenase. Further preferred recombinant DNAs according to the invention contain promoters of ribosomal protein encoding genes, more preferably wherein the transcription promoter comprises a region found upstream of the open reading frame encoding a protein as represented by one of the disclosed amino acid sequences. According to a further aspect of the invention an isolated DNA sequence coding for an enzyme involved in the carotenoid biosynthetic pathway of Phaffia rhodozyma is provided.Type: GrantFiled: November 19, 1998Date of Patent: December 11, 2001Assignee: DSM N.V.Inventors: Albert Johannes Joseph Van Ooijen, Jan Cornelis Verdoes, Jan Wery
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Isopentenyl diphosphate isomerase from Hevea brasiliensis and rubber producing method using the same
Patent number: 6316695Abstract: The present invention cloned a cDNA clone encoding isopentenyl diphosphate (hereafter “IPP”) isomerase (EC 5.3.3.2) from a cDNA library of Hevea brasiliensis latex. The clone has a continuous open reading frame encoding a peptide of 234 amino acids with a predicted molecular mass of 26.7 kDa. The deduced protein is acidic with an isoelectric point of 4.7 and shows high sequence identity with other IPP isomerases. The recombinant protein expressed in Escherichia coli showed IPP isomerase activity. In vitro rubber biosynthesis assays using washed rubber particle (WRP) deprived of initiating allylic diphosphates were performed with the addition of IPP isomerase in the reaction mixture. Results revealed that the recombinant IPP isomerase is catalytically active in catalyzing the conversion of IPP to DMAPP, a key activation step of the basic five-carbon isoprene unit in rubber biosynthesis. Southern analysis indicated that the IPP isomerase is encoded by two genes in Hevea rubber tree.Type: GrantFiled: April 22, 1999Date of Patent: November 13, 2001Assignee: Korea Kumho Petrochemical Co., Ltd.Inventors: Kyung-Han Han, Hun-Seung Kang, Soo-Kyung Oh, Dong-Ho Shin, Jae-Mo Yang -
Publication number: 20010031461Abstract: A method for potentiating a host cell for sensitivity to a heterologous polynucleotide comprises providing a population of host cells, wherein the cells comprise a heterologous polynucleotide, a first promoter regulating the expression of the heterologous polynucleotide, and a replicable genetic element comprising a second polynucleotide encoding a repressor of the first promoter, a reporter gene under the control of a second promoter, expression of which provides a detectable label, and an origin of replication, wherein the replicable genetic element is subject to loss by the host cells, thus resulting in heterogeneously labeled colonies when the host cells are cultured; mutagenizing the population of host cells; growing colonies of the mutagenized host cells under conditions wherein the heterologous polynucleotide, the repressor, and the reporter are all expressed; and identifying a colony of mutagenized host cells that is homogeneously labeled.Type: ApplicationFiled: December 15, 2000Publication date: October 18, 2001Inventor: Stuart Tugendreich
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Patent number: 6300095Abstract: The invention relates to promoters of the genes glutamate dehydrogenase, &bgr;-acetylhexosaminidase and &ggr;-actin and their use in systems of expression, secretion and anti-sense of filamentary fungi. The invention also relates to the use of the promoters of the genes which code: (I) glutamate dehydrogenase NADP depending (EC.1.4.1.4) of Penicillium chrysogenum, (II) &ggr;-N-actylhexosaminidase (EC.3.2.1.52) of Penicillium chrysogenum and (III) &ggr;-actin of Penicillium chrysogenum and Acrimonium chrysogenum, which can be used for the construction of potent vectors of expression and secretion useful both for P. chrysogenum and for A. chrysogenum and related species. These promoters can also be used for blocking the genic expression through anti-sense construction. Under the control of the above mentioned promoters, it is possible to conduct the expression of other genes in filamentary fungi, thereby increasing the production of antibiotics and/or proteins inherent to the same.Type: GrantFiled: May 14, 1999Date of Patent: October 9, 2001Assignee: Antibioticos, S.A.Inventors: Jose Luis Barredo Fuente, Marta Rodriguez Saiz, Alfonso J. Collados De La Vieja, Migeul Angel Moreno Valle, Francisco Salto Maldonado, Bruno Diez Garcia
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Intrabodies with defined framework that is stable in a reducing environment and applications thereof
Publication number: 20010024831Abstract: A method for the isolation of CDRs in a defined framework that is stable and soluble in reducing environment is described as well as thus obtainable scFv. Starting from such scFv with defined framework a scFv library can be generated wherein the framework is conserved while at least one complementary determining region (CDR) is randomized. Such library, e.g. in yeast cells, is suitable for screening for antibody/CDR-interactions or for screening for antibodies.Type: ApplicationFiled: December 28, 2000Publication date: September 27, 2001Inventors: Adrian Auf Der Maur, Alcide Barberis, Dominik Escher -
Patent number: 6280937Abstract: The invention provides shuttle vectors, and methods of using shuttle vectors, capable of expression in, at least, a mammalian cell. Furthermore, the shuttle vectors are capable of replication in at least yeast, and optionally, bacterial cells. Also provided is a method wherein yeast are transformed with a shuttle vector as provided herein. Heterologous nucleic acids flanked by 5′ and 3′ ends identical to a homologous recombination site within the shuttle vector are introduced to the transformed yeast and allowed to homologously recombine with the shuttle vector such that they are inserted into the vector by the yeast organism. The shuttle vector is then recovered and transferred to a mammalian cell for expression.Type: GrantFiled: August 14, 1998Date of Patent: August 28, 2001Assignee: Rigel Pharmaceuticals, Inc.Inventors: Ying Luo, Pei Wen Yu, James Lorens
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Patent number: 6277631Abstract: Recombinant yeast expression vectors with the features indicated in the patent claims are described. These recombinant yeast expression vectors can be used for the preparation of HBeAg in yeast host organisms. Appropriate expression systems, transformed host organisms, diagnostic aids and medicinal agents are additionally described.Type: GrantFiled: May 22, 1995Date of Patent: August 21, 2001Assignee: Dade Behring Marburg GmbHInventors: Michael Noah, Michael Bröker
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Patent number: 6265186Abstract: The present invention provides for a yeast cell comprising at least two copies of a desired gene integrated into its chromosomal genome, wherein said genome comprises at least two DNA domains suitable for integration of one or more copies of said desired gene, which domains share substantial sequence homology and are non-ribosomal RNA encoding DNA domains, and wherein at least two of said substantially homologous non-ribosomal RNA encoding DNA domains have at least one copy of the said desired gene integrated. The invention also provides methods for making yeast cells according to the invention, as well as the use thereof for making a protein, a peptide or a metabolite.Type: GrantFiled: December 10, 1999Date of Patent: July 24, 2001Assignee: DSM N.V.Inventors: Bart Willem Swinkels, Albert Johannes Joseph Van Ooijen, Adriana Cornelia Maria Noordermeer-Van Der Haak
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Patent number: 6265185Abstract: Novel yeast promoters for either EF1-alpha protein or ribosomal protein S7 gene suitable for expression cloning in yeast and heterologous expression of proteins in yeast. The yeast promoters are preferably active in the pH range 4-11 without peptone and obtained from the yeast strain Yarrowia lipolytica.Type: GrantFiled: October 19, 1998Date of Patent: July 24, 2001Assignee: Novozymes A/S PatentsInventors: Sven Muller, Henrik Dalbøge
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Patent number: 6232074Abstract: 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.Type: GrantFiled: December 10, 1999Date of Patent: May 15, 2001Assignee: CompleGen, Inc.Inventors: Dean Dawson, John Swindle
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Patent number: 6228583Abstract: Methods of identifying agents or compounds which are capable of inhibiting the replication and/or accumulation of DNA circles in cells are described. Also described are methods of assessing the ability of a compound to extend life span, as well as methods of extending life span, comprising administering to a cell a compound identified by the assays described herein which extends life span. The invention also pertains to isolated mWRN, or an active derivative or fragment thereof and to an isolated nucleic acid molecule which encodes mWRN, or an active derivative or fragment thereof.Type: GrantFiled: July 31, 1998Date of Patent: May 8, 2001Assignee: Massachusetts Institute of TechnologyInventors: Leonard P. Guarente, David A. Sinclair
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Patent number: 6221588Abstract: The functional analysis of genes frequently requires the manipulation of large genomic regions. A yeast-bacteria shuttle vector is described, that can be used to clone large regions of DNA by homologous recombination. The important feature of present invention is the presence of the a bacterial replication origin, which allows large DNA insert capacity. The utility of this vector lies in its ability to isolate, manipulate and maintain large fragments in bacteria and yeast, allowing for mutagenesis by yeast genetics and simplified preparation of plasmid DNA in bacteria.Type: GrantFiled: June 10, 1998Date of Patent: April 24, 2001Assignee: Yale UniversityInventors: M. Suzanne Bradshaw, Jacques A. Bollekens, Frank H. Ruddle
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Patent number: 6221630Abstract: The invention provides methods and recombinant expression constructs for inducing and sustaining high-level production of a recombinant polypeptide in yeast. The invention specifically provides high copy number recombinant expression constructs that express high levels of trans-acting transcription factors that in turn induce expression of a recombinant nucleic acid encoding a heterologous or endogenous recombinant polypeptide. The invention more specifically provides constructs that express galactose-inducible and temperature-sensitive transcription factors. The invention also provides constructs comprising nucleic acids the transcription of which is regulated by the transcription factors expressed by the construct. The invention also provides yeast cells transformed by the recombinant expression constructs of the invention that permit sustained high-level expression of a recombinant polypeptide.Type: GrantFiled: March 24, 1999Date of Patent: April 24, 2001Assignee: The Penn State Research FoundationInventor: James E. Hopper
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Patent number: 6218145Abstract: A composition for expression of a protein-encoding gene in a host cell is described, making use of an heterologous regulon. This composition provides a protein-encoding gene under control of a promoter heterologous to the host cell, and a gene for an RNA polymerase, preferably a single-subunit RNA polymerase that recognizes the promoter heterologous to the host cell. The gene for the RNA polymerase is under control of an inducible promoter recognized by the host cell. Also disclosed is a method for expressing the protein-encoding gene using the composition described.Type: GrantFiled: April 1, 1999Date of Patent: April 17, 2001Assignee: Monsanto CompanyInventors: Gregg Bogosian, Julia P. O'Neil, Jeffrey M. Staub
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Patent number: 6214577Abstract: A vector having a gene for resistance to an antibiotic otherwise capable of killing a host yeast cell, the gene being transcribed from a yeast promoter sequence.Type: GrantFiled: June 6, 1995Date of Patent: April 10, 2001Inventor: Robert Rogers Yocum
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Patent number: 6215044Abstract: Disclosed are nucleic acids from tomato encoding polypeptides having xyloglucan endo-transglycosylase (XET) activity, comprising residues 21-289 of the amino acid sequence shown in FIG. 1 (SEQ ID NO:2) or or comprising residues 19-287 of the amino acid sequence shown in FIG. 5 (SEQ ID NO:8), and transgenic plants comprising said nucleic acids.Type: GrantFiled: May 6, 1996Date of Patent: April 10, 2001Assignee: Unilever Patent Holdings B.V.Inventors: David A. Arrowsmith, Susan A. Hellyer, Jacqueline de Silva, Sally A. Whiteman
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Patent number: 6194169Abstract: Enhanced yields of the platelet-derived growth factor (PDGF) B-chain are obtained using the Pichia pastoris yeast system. Yields of both wild-type and mutant human proteins are enhanced when the yeast is transformed with a vector containing the yeast mating factor promoter fused to a sequence encoding the mature protein. The secreted wild-type protein is indistinguishable from mature 29-32 kDa protein isolated from human and Chinese hamster ovary (CHO) cells. An RKK→EEE mutant exhibited reduced association with the cell surface and accumulated in the culture medium as 29-32 kDa forms. Stable transfection of U87 astrocytoma cells with RKK→EEE mutants of either the A- or B-chain inhibited malignant growth in athymic nude mice. Despite altered receptor binding activities, each mutant retained full mitogenic activity when applied to cultured Swiss 3T3 cells. Circular dichroism spectrophotometric analysis of the RKK→EEE mutant revealed a secondary structure indistinguishable from the wild type.Type: GrantFiled: June 4, 1998Date of Patent: February 27, 2001Assignee: The University of Kentucky Research FoundationInventor: David M. Kaetzel
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Patent number: 6190914Abstract: The invention provides methods for changing the metabolic pathways of micro-organisms in the presence of a certain carbon source and uses of such changes, as well as micro-organisms and uses of such changes, as well as micro-organisms produced by these methods. In a preferred embodiment the invention provides new yeast strains with improved biomass yields, a process to obtain these yeasts and the potential application of these yeasts are provided. The biomass yield is improved by the introduction into a yeast of a DNA construct conferring altered expression of a gene encoding a protein controlling transcription of a number of glucose-repressed genes. The yeasts are less sensitive to glucose repression, resulting in an increase in respiratory capacity, reduction of ethanol production and increased conversion of sugar into biomass.Type: GrantFiled: June 14, 1999Date of Patent: February 20, 2001Assignee: Universiteit van AmsterdamInventors: Leslie Alan Grivell, Maarten Joost Teixeira De Mattos, Jolanda Blom
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Patent number: 6146867Abstract: The present invention provides the complete cDNA sequence of maize acetyl CoA carboxylase and a method introducing and expressing a plant acetyl CoA carboxylase gene in plant cells. The method includes the steps of introducing an expression cassette encoding a plant acetyl CoA carboxylase or an antisense DNA sequence complementary to the sequence for a plant acetyl CoA carboxylase gene operably linked to a promoter functional in plant cells, into the cells of a plant tissue and expressing the plant acetyl CoA carboxylase gene. The expression cassette can also be introduced into other host cells to increase yield of a plant acetyl CoA carboxylase crystallized enzyme.Type: GrantFiled: August 12, 1996Date of Patent: November 14, 2000Assignees: Regents of the University of Minnesota, United States of AmericaInventors: Burle G. Gengenbach, David A. Somers, Donald L. Wyse, John W. Gronwald, Margaret A. Egli, Sheila M. Lutz
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Patent number: 6100042Abstract: Yeast cells are engineered to express both a surrogate of a pheromone system protein (e.g., enzymes involved in maturation of .alpha.-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.Type: GrantFiled: October 13, 1994Date of Patent: August 8, 2000Assignee: Cadus Pharmaceutical CorporationInventors: Dana Merriman Fowlkes, Jim Broach, John Manfredi, Christine Klein, Andrew J. Murphy, Jeremy Paul, Joshua Trueheart
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Patent number: 6069298Abstract: The present invention provides the complete cDNA sequence of maize acetyl CoA carboxylase and methods for conferring herbicide tolerance and/or altering the oil content of plants by introducing and expressing a plant acetyl CoA carboxylase gene in plant cells. The method of imparting herbicide tolerance to a plant includes the steps of introducing an expression cassette encoding a plant acetyl CoA carboxylase or an antisense DNA sequence complementary to the sequence for a plant acetyl CoA carboxylase gene operably linked to a promoter functional in plant cells, into the cells of a plant tissue and expressing the plant acetyl CoA carboxylase gene in an amount effective to render the acetyl CoA carboxylase and/or plant cell substantially tolerant to the herbicides. The method of altering the oil content in a plant includes the steps of introducing an expression cassette into plant cells and expressing the acetyl CoA carboxylase gene in an amount effective to alter the oil content of the cells.Type: GrantFiled: April 5, 1995Date of Patent: May 30, 2000Assignees: Regents of the University of Minnesota, The United States of AmericaInventors: Burle G. Gengenbach, David A. Somers, Donald L. Wyse, John W. Gronwald, Margaret A. Egli, Sheila M. Lutz
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Patent number: 6066715Abstract: Disclosed are new ligands for use in a binding assay for proteases and phosphatases, which contain cysteine in their binding sites or as a necessary structural component for enzymatic binding. The sulfihydryl group of cysteine is the nucleophilic group in the enzyme's mechanistic proteolytic and hydrolytic properties. The assay can be used to determine the ability of new, unknown ligands and mixtures of compounds to competitively bind with the enzyme versus a known binding agent for the enzyme, e.g., a known enzyme inhibitor. By the use of a mutant form of the natural or native wild-type enzyme, in which serine, or another amino acid, e.g., alanine, replaces cysteine, the problem of interference from extraneous oxidizing and alkylating agents in the assay procedure is overcome. The interference arises because of oxidation or alkylation of the sulfihydryl, --SH (or --S.sup.-), in the cysteine, which then adversely affects the binding ability of the enzyme.Type: GrantFiled: November 4, 1997Date of Patent: May 23, 2000Assignee: Merck Frosst Canada & Co.Inventors: Sylvie Desmarais, Richard Friesen, Robert Zamboni
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Patent number: 6060249Abstract: The present invention relates to a novel method of identifying cDNA's which encode secreted and membrane-bound proteins. The present invention also relates to a novel method for preparing cDNA libraries enriched for signal sequences. The methods of the invention provide for an improved signal sequence detection system which results, when compared to the prior art, in a greater number of correctly identified signal sequences and less total time required to complete the procedure.Type: GrantFiled: February 24, 1998Date of Patent: May 9, 2000Assignee: Genentech, Inc.Inventors: Kevin Baker, Austin Gurney
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Patent number: 6051431Abstract: 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.Type: GrantFiled: December 8, 1997Date of Patent: April 18, 2000Assignee: DSM N.V.Inventors: Gerardus Cornelius Maria Selten, Bart Willem Swinkels, Robertus Franciscus Maria Van Gorcom
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Patent number: 6033883Abstract: Hybrid and novel polyketide synthases and polyketides are produced by use of a multiple vector system. The combinatorial possibilities offered by placing the various catalytic activities of PKS systems on separate vectors permits the construction of improved libraries of PKS and polyketides. In addition, polyketides can be produced in hosts that ordinarily do not produce polyketides by supplying, along with an expression system for the desired PKS, an expression system for holo ACP synthase.Type: GrantFiled: December 11, 1997Date of Patent: March 7, 2000Assignee: KOSAN Biosciences, Inc.Inventors: Philip J. Barr, Daniel V. Santi, Gary W. Ashley, Rainer Ziermann
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Patent number: 6031155Abstract: A genomic nucleic acid sequence encoding a 62 kDa barley endoxylanase has been isolated and characterized. The genomic DNA sequences are used to transform plant cells for expression of enhanced amounts of active endoxylanase.Type: GrantFiled: June 5, 1997Date of Patent: February 29, 2000Inventors: Verena Cameron-Mills, Finn Lok, Catharina Maria Cornelia Sinjorgo, Ronald Tako Marinus van den Dool, Martinus Petrus Maria Caspers, Maria Joanna van Zeijl-van der Valk
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Patent number: 6022712Abstract: An isolated DNA segment is disclosed which is derived from a Saccharomyces cerevisiae sorbitol dehydrogenase gene and which functions to increase expression of an associated foreign polypeptide when the DNA segment and the gene coding for the foreign polypeptide are operably linked in a vector in such a manner that the vector is replicated and carried by a host yeast cell. The functionally active portion of the segment is under the control (i) a transcriptional regulatory sequence of the sorbitol dehydrogenase gene of Saccharomyces cerevisiae, (ii) a translation initiation regulatory sequence of the sorbitol dehydrogenase gene of Saccharomyces cerevisiae, and (iii) a termination regulatory sequence of the sorbitol dehydrogenase gene of Saccharomyces cerevisiae. Yeast cells containing these regulatory sequences linked to a foreign DNA sequence are grown in medium containing sorbitol under conditions permitting a foreign polypeptide to be expressed.Type: GrantFiled: June 6, 1995Date of Patent: February 8, 2000Assignee: Abbott LaboratoriesInventors: Aparna V. Sarthy, Cynthia W. Schopp
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Patent number: 6013472Abstract: The invention relates to the DNA and protein encoded by the GA4 locus. This protein is believed to be a member of the family of enzymes involved in the biosynthesis of the gibberellin family (GA) of plant growth hormones which promote various growth and developmental processes in higher plants, such as seed germination, stem elongation, flowering and fruiting. More specifically, the protein encoded by the GA4 locus is an hydroxylase. The invention also relates to vectors containing the DNA and the expression of the protein encoded by the DNA of the invention in a host cell. Additional aspects of the invention are drawn to host cells transformed with the DNA or antisense sequence of the invention, the use of such host cells for the maintenance, or expression or inhibition of expression of the DNA of the invention and to transgenic plants containing DNA of the invention. Finally, the invention also relates to the use of the protein encoded by the GA4 locus to alter aspects of plant growth.Type: GrantFiled: January 17, 1997Date of Patent: January 11, 2000Assignee: The General Hospital CorporationInventors: Hui-Hwa Chiang, Howard M. Goodman
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Patent number: 6004779Abstract: 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.Type: GrantFiled: August 21, 1998Date of Patent: December 21, 1999Assignee: Scriptgen Pharmaceuticals, Inc.Inventors: John D. Bradley, Craig M. Thompson, Jeffrey B. Moore, C. Richard Wobbe, Judith M. Healy, Caroline E. Donnelly
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Patent number: 5972708Abstract: A system is described which utilizes a novel system of repressor titration for maintenance of a plasmid useful in gene therapy and production of a recombinant protein. The system utilizes a transformed host cell containing a plasmid including an operator susceptible to binding by a repressor expressed in trans, a first chromosomal gene encoding the repressor, and a second chromosomal gene that is functionally associated with an operator and essential for cell growth, wherein the plasmid is present in the cell in sufficient numbers to titrate the repressor such that the essential gene is expressed, thereby permitting cell growth.Type: GrantFiled: October 21, 1998Date of Patent: October 26, 1999Assignee: Cobra Therapeutics LimitedInventors: David J. Sherratt, Steven G. Williams, Julian A.J. Hanak
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Patent number: 5965444Abstract: A DNA construct wherein a DNA fragment which is recombinable in yeast chromosomal DNA is directly or indirectly linked at both ends of a DNA fragment which comprises a pair of R sensitive sequences oriented in the same direction and flaking both an R gene placed under the control of an inducible promoter and an expressible selective marker gene, which is a DNA construct designed with the R sensitive sequences non-symmetrically shortened, so that no functionable R sensitive sequence remains after the R sensitive sequence recombination has occurred by expression of the R gene and the selective marker has been removed. Since no functionable R sensitive sequence remains after removal of the selective marker, recombination does not occur again, and thus the same selective marker may be used for multiple insertions of foreign genes.Type: GrantFiled: June 23, 1997Date of Patent: October 12, 1999Assignee: Suntory LimitedInventors: Toshihiko Ashikari, Hiroto Kondo, Keiko Sakakibara, Hiroyuki Araki, Yasuji Oshima
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Patent number: 5948620Abstract: The invention relates to modified two-hybrid systems, in particular a reverse two-hybrid system which employs as a reporter a gene encoding a modifying agent such as an enzyme, and a signal agent which is modified by the enzyme usually by being broken down, such that in the event of an inhibition of binding of the two hybrid proteins a detectable signal is produced. The system is particularly useful for drug screening.Type: GrantFiled: August 4, 1997Date of Patent: September 7, 1999Assignee: Amersham International PLCInventors: Douglas Hurd, Rachel Alison Fallon, Nicholas Ian Workman, Susan Jane Dale
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Patent number: 5948682Abstract: The present invention relates to the use of a class of genes called oil body protein genes that have unique features. The discovery of these features allowed the invention of methods for the production of recombinant proteins wherein a protein of interest can be easily separated from other host cell components. The invention is further exemplified by methods for exploitation of the unique characteristics of the oil body proteins and oil body genes for expression of polypeptides of interest in many organisms, particularly plant seeds. Said polypeptides may include but are not limited to: seed storage proteins, enzymes, bioactive peptides, antibodies and the like. The invention can also be modified to recover recombinant polypeptides fused to oil body proteins from non-plant host cells. Additionally the invention provides a method of using recombinant proteins associated with seed oil bodies released during seed germination for expression of polypeptides that afford protection to seedlings from pathogens.Type: GrantFiled: April 25, 1997Date of Patent: September 7, 1999Assignee: Sembiosys Genetics Inc.Inventor: Maurice M. Moloney
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Patent number: 5945339Abstract: The invention concerns genes encoding recombinases that can be used to promote homologous recombination in eukaryotic cells. The application teaches methods by which a recombinase of one species can be used to isolate a homologous recombinase of a different species and methods to identify the isolated homologs. Recombinases from Ustilago maydis, Saccharomyces cerevisiae and humans are specifically included in the invention.The invention encompasses the method of producing an isolated recombinase protein in a prokaryotic cell and recovering the product in an active form. The invention also encompasses a genetically engineered gene which encodes a non-naturally occurring recombinase that causes a greater rate of recombination than does the naturally occurring recombinase. The invention further encompasses the use of recombinase proteins and of recombinase genes to promote homologous recombination, including recombination between a host cell genome and a chimeric oligonucleotide, i.e.Type: GrantFiled: July 13, 1998Date of Patent: August 31, 1999Assignees: Cornell Research Foundation, Inc., Thomas Jefferson UniversityInventors: William K. Holloman, Eric B. Kmiec
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Patent number: 5945329Abstract: The invention provides customized proteases (i.e., mutant enzymes), methods of making customized proteases, as well as methods of using customized proteases. The customized proteases of the invention are derived from the known proteases. Altered transacylation reactions include the capability to perform transacylation reactions not substantially catalyzed by the known protease or the capability to perform transacylation reactions with improved yields, or both. The methods of the invention provide for customized proteases through site specific or random mutagenesis of the active site amino acids of the known proteases. The invention also provides for methods of using the customized proteases to prepare a preselected transacylation products. The preselected transacylation products produced can be modified by substitution at the N- or C-terminal with nucleophiles such as L-amino acids, D-amino acids, amino acid amides, and radioactive amino acids.Type: GrantFiled: July 23, 1997Date of Patent: August 31, 1999Assignee: Carlsberg A/SInventors: Klaus Breddam, Morten C. Kielland-Brandt, Uffe Hasbo Mortensen, Kjeld Ove Olesen, Henning Ralf Stennicke, Fred W. Wagner
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Patent number: 5928868Abstract: Methods and a kit are provided for characterizing small molecules from a library of small molecules or alternatively identifying protein targets to which known small molecules bind. The methods include forming hybrid ligand in which at least one ligand is a small molecule. The hybrid ligand is introduced into cells that in turn contain a first and a second expression vector. Each expression vector includes DNA for expressing a hybrid protein that encodes a target protein linked to a coding sequence for a transcriptional module. The cells further contains a reporter gene, the expression of which is conditioned on the proximity of the first and second hybrid proteins to each other, an event that occurs only if the hybrid ligand binds to target sites on both hybrid proteins. Those cell which express the reporter gene are selected and the unknown small molecule or the unknown hybrid protein is identified.Type: GrantFiled: April 25, 1997Date of Patent: July 27, 1999Assignee: Massachusetts Institute of TechnologyInventors: Jun Liu, Edward J. Licitra
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Patent number: 5919651Abstract: DNA expression vectors capable, in a transformant strain of yeast, of expressing a polypeptide under the control of a genetically distinct yeast promoter, processes of forming transformant strains of yeast and transformed yeast strains are disclosed.Type: GrantFiled: June 6, 1995Date of Patent: July 6, 1999Assignees: Washington Research Foundation, Genentech, Inc.Inventors: Ronald A. Hitzeman, Franklin E. Hagie, IV, Benjamin D. Hall, Gustav Ammerer
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Patent number: 5885799Abstract: The protease necessary for polyprotein processing in Hepatitis C virus is identified, cloned, and expressed. Proteases, truncated protease, and altered proteases are disclosed which are useful for cleavage of specific polypeptides, and for assay and design of antiviral agents specific for HCV.Type: GrantFiled: September 6, 1996Date of Patent: March 23, 1999Assignee: Chiron CorporationInventors: Michael Houghton, Qui-Lim Choo, George Kuo
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Patent number: 5856123Abstract: DNA expression vectors capable, in a transformant strain of yeast, of expressing a polypeptide under the control of a genetically distinct yeast promoter, processes of forming transformant strains of yeast and transformed yeast strains are disclosed.Type: GrantFiled: June 6, 1995Date of Patent: January 5, 1999Assignees: Washington Research Foundation, Genentech, Inc.Inventors: Ronald A. Hitzeman, Franklin E. Hagie IV, Benjamin D. Hall, Gustav Ammerer
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Patent number: 5854018Abstract: DNA expression vectors capable, in a transformant strain of yeast, of expressing a polypeptide under the control of a genetically distinct yeast promoter, processes of forming transformant strains of yeast and transformed yeast strains are disclosed.Type: GrantFiled: June 6, 1995Date of Patent: December 29, 1998Assignees: Washington Research Foundation, Genentech, Inc.Inventors: Ronald A. Hitzeman, Franklin E. Hagie IV, Benjamin D. Hall, Gustav Ammerer
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Patent number: RE37343Abstract: A yeast &agr;-factor expression system is provided comprised of a truncated leader sequence, containing the &agr;-factor signal peptide and one glycosylation site, linked by a processing site to a non-yeast protein-encoding sequence.Type: GrantFiled: August 27, 1998Date of Patent: August 28, 2001Assignee: Chiron CorporationInventor: Patricia Tekamp-Olson