Yeast Is A Host For The Plasmid Or Episome Patents (Class 435/483)
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Publication number: 20030077707Abstract: Combinatorial libraries of polyketides can be obtained by suitable manipulation of a host modular polyketide synthase gene cluster such as that which encodes the PKS for erythromycin. The combinatorial library is useful as a source of pharmaceutically active compounds.Type: ApplicationFiled: March 12, 2002Publication date: April 24, 2003Inventors: Chaitan Khosla, Gary Ashley, Hong Fu, Camilla M. Kao, Robert McDaniel
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Publication number: 20030077831Abstract: Compositions and methods for expression of heterologous mammalian proteins and their secretion in the biologically active mature form using a yeast host cell as the expression system are provided. Compositions of the invention are nucleotide sequences encoding a signal peptide sequence for a yeast secreted protein, an optional leader peptide sequence for a yeast secreted protein, a native propeptide leader sequence for a mature protein of interest, and a sequence for the mature protein of interest, all operably linked to a yeast promoter. Each of these elements is associated with a processing site recognized in vivo by a yeast proteolytic enzyme. Any or all of these processing sites may be a preferred processing site that has been modified or synthetically derived for more efficient cleavage in vivo. The compositions are useful in methods for expression of heterologous mammalian proteins and their secretion in the biologically active mature form.Type: ApplicationFiled: October 25, 2002Publication date: April 24, 2003Applicant: Chiron CorporationInventor: Patricia Tekamp-Olson
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Patent number: 6551786Abstract: A method for selecting compounds capable of modulating protein-protein interactions is provided, in which two fusion proteins are prepared and allowed to interact with each other in the presence of test compounds. The interaction between the two fusion proteins leads to protein trans-splicing, producing an active reporter. Compounds that disrupt or enhance protein-protein interactions can be selected based on the presence or absence of the active reporter.Type: GrantFiled: January 4, 2002Date of Patent: April 22, 2003Assignee: Myriad Genetics, Inc.Inventor: John Manfredi
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Publication number: 20030073610Abstract: Screening methods for identifying substances that provide therapeutic value for various diseases associated with protein misfolding are provided. Genetic and chemical screening methods are provided using a yeast system. The methods of the invention provide a rapid and cost-effective method to screen for compounds that prevent protein misfolding and/or protein fibril formation and/or protein aggregation which includes numerous neurodegenerative diseases including Parkinson's disease, Alzheimer's disease, Huntington's disease as well as non-neuronal diseases such as type 2 diabetes.Type: ApplicationFiled: February 15, 2002Publication date: April 17, 2003Applicant: The University of ChicagoInventors: Susan Lindquist, Sylvia Krobitsch, Tiago Fleming Outeiro
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Publication number: 20030065159Abstract: 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.Type: ApplicationFiled: July 16, 2002Publication date: April 3, 2003Applicant: Genentech, Inc.Inventors: Kevin P. Baker, Jian Chen, Luc Desnoyers, Audrey Goddard, Paul J. Godowski, Austin L. Gurney, James Pan, Victoria Smith, Colin K. Watanabe, William I. Wood, Zemin Zhang
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Publication number: 20030064436Abstract: The invention relates to a method for producing hydroxylated triple helical proteins in yeast host cells by introducing to a suitable yeast host cell, DNA sequences encoding the triple helical protein as well as prolyl 4-hydroxylase (PH4), in a manner wherein the introduced DNA sequences are replicated, stably retained and segregated by the yeast host cells.Type: ApplicationFiled: December 21, 2001Publication date: April 3, 2003Inventors: Paul Richard Vaughan, Maria Galanis, John Alan Maurice Ramshaw, Jerome Anthony Werkmeister
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Patent number: 6541622Abstract: The present invention provides the promoter clone discovery of an alpha-amylase gene of a starch utilizing yeast strain Schwanniomyces castellii. The isolated alpha-amylase promoter is an inducible promoter, which can regulate strong gene expression in starch culture medium.Type: GrantFiled: August 4, 2000Date of Patent: April 1, 2003Assignee: Battelle Memorial InstituteInventors: Johnway Gao, Rodney S. Skeen, Brian S. Hooker, Daniel B. Anderson
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Patent number: 6534315Abstract: The present invention relates to a DNA cassette intended for the transformation of yeast, leaving no useless exogenous DNA but the gene(s) of interest comprising at least one negative dominant marker, two direct repeat sequences (DRS) which are non exogenous and non recombinogenic with the genome of the host strain, these two direct repeat sequences flanking the negative dominant marker and optionally at least one gene of interest containing, if necessary, the elements necessary for its expression in the host cell. The invention relates as well to a method of integration of gene(s) of interest or inactivation of a gene in yeast, and of transformation of yeast with the DNA cassette, and to yeast strains thus obtained.Type: GrantFiled: October 12, 1999Date of Patent: March 18, 2003Assignee: La Societe Lesaffre et CieInventors: Jürgen Bauer, Valérie Nacken, Annie Loiez
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Publication number: 20030049785Abstract: A new expression vector for the production of a polypeptide in yeast. The vector includes a sequence coding for the polypeptide and other sequences allowing expression of the polypeptide only in yeast. The other sequences lack any non-yeast sequences. Other embodiments include a yeast strain comprising such a vector, a method for the production of the vector, a method for the production of the yeast strain by transformation of a yeast strain with the new vector, and a method for the production of a polypeptide in the transformed yeast strain by fermentation thereof followed by isolation of the polypeptide.Type: ApplicationFiled: September 3, 2002Publication date: March 13, 2003Applicant: Patentpharm AG.Inventors: Thomas Schreier, Rainer Voegeli
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Publication number: 20030050459Abstract: 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.Type: ApplicationFiled: July 15, 2002Publication date: March 13, 2003Applicant: Genentech, Inc.Inventors: Kevin P. Baker, Jian Chen, Luc Desnoyers, Audrey Goddard, Paul J. Godowski, Austin L. Gurney, James Pan, Victoria Smith, Colin K. Watanabe, William I. Wood, Zemin Zhang
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Publication number: 20030049822Abstract: Novel genes have been isolated which encode cytochrome P450 and NADPH reductase enzymes of the &ohgr;-hydroxylase complex of C. tropicalis 20336. Vectors including these genes, transfected host cells and transformed host cells are provided. Methods of producing of cytochrome P450 and NADPH reductase enzymes are also provided which involve transforming a host cell with a gene encoding these enzymes and culturing the cells. Methods of increasing the production of a dicarboxylic acid and methods of increasing production of the aforementioned enzymes are also provided which involve increasing in the host cell the number of genes encoding these enzymes. A method for discriminating members of a gene family by quantifying the expression of genes is also provided.Type: ApplicationFiled: May 3, 2002Publication date: March 13, 2003Inventors: C. Ron Wilson, David L. Craft, L. Dudley Eirich, Mark Eshoo, Krishna M. Madduri, Cathy A. Cornett, Alfred A. Brenner, Maria Tang, John C. Loper, Martin Gleeson
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Patent number: 6531289Abstract: The invention provides novel yeast cells comprising genes whose expression can be modulated by growth in the presence or absence of metal ions, methods for making such yeast cells, and methods of using such yeast cells for determining the requirement for expression of particular genes for the growth or viability of the yeast cells. The invention also provides methods of using such yeast cells in the isolation, screening and analysis of candidate antifungal compounds.Type: GrantFiled: May 18, 2000Date of Patent: March 11, 2003Assignee: Anadys Pharmaceuticals, Inc.Inventors: John D. Bradley, Craig M. Thompson, Jeffrey B. Moore, C. Richard Wobbe, David A. Bailey
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Publication number: 20030044866Abstract: This patent describes a novel method of detecting genetic interactions in yeast. This method can also be used to screen for function of biological effectors on yeast. The method encompasses crossing yeast strains with genetic alterations to acquire double mutants. The phenotypes of these double mutants are then checked to detect genetic interactions between the double mutants. This method can be used to assign function to yeast genes and their viral, prokaryotic, and eukaryotic homologs, and aptamers. It can also be used to study yeast two hybrid interactions and to find genes that regulate certain yeast promoters.Type: ApplicationFiled: August 15, 2001Publication date: March 6, 2003Inventor: Charles Boone
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Publication number: 20030040047Abstract: The present invention relates to methods for producing recombinant proteins, in particular recombinant secretory proteins, to a method for identifying nucleic acid molecules the expression products of which permit improved secretion of a recombinant secretory protein, to the use of molecules thus identified for enhancing secretion of heterologous proteins, and to corresponding kit systems.Type: ApplicationFiled: October 31, 2001Publication date: February 27, 2003Applicant: Rhein BiotechInventors: Mike Farwick, Markus London, Juergen Dohmen, Ulrike Dahlems, Gerd Gellissen, Alexander W. Strasser
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Publication number: 20030039989Abstract: The present invention provides a method for constructing an expression library containing a gene that is toxic to bacteria, comprising the steps of: (a) isolating genomic DNA from a culture of cells of a first bacterium, wherein said genomic DNA contains a gene toxic to a second bacterium; (b) partially digesting said genomic DNA to produce genetic inserts, wherein at least one of the genetic inserts contains a gene to said second bacterium; (c) cloning said genetic inserts into a cloning vector; (d) ligating together, under suitable ligation conditions, said genetic inserts and said cloning vector, to produce ligation products; (e) transforming said ligation products directly into yeast; and (f) amplifying said ligation products in said yeast. Also provided are an expression library constructed in accordance with this method and a method for screening for a gene that is toxic to bacteria using said expression library.Type: ApplicationFiled: December 17, 2001Publication date: February 27, 2003Inventors: Steven Arthur Haney, Piet A. J. de Boer
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Publication number: 20030022280Abstract: The present invention relates to compositions and methods for producing high yields of heterologous polypeptides in a Pichia cell such as Pichia methanolica are of interest.Type: ApplicationFiled: March 9, 2001Publication date: January 30, 2003Applicant: Novozymes A/SInventors: Shinobu Takagi, Yuji Terui, Noriko Tsutsumi, Rikako Taira
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Publication number: 20030017601Abstract: 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: ApplicationFiled: January 7, 2002Publication date: January 23, 2003Applicant: Rigel Pharmaceuticals, Inc.Inventors: Ying Luo, Pei Wen Yu, James Lorens
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Patent number: 6503749Abstract: A subject of the invention is a modified yeast strain in which the acetyl-CoA pregnenolone acetyltransferase (APAT) activity is eliminated by altering the gene coding for this activity with resultant stabilization of 3&bgr;-hydroxysteroids.Type: GrantFiled: March 1, 2001Date of Patent: January 7, 2003Assignee: Aventis Pharma S.A.Inventors: Tilman Achstetter, Gilles Cauet, Eric Degryse
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Patent number: 6503712Abstract: The present invention is directed to methods for producing gene targeting constructs by homologous recombination using mouse genomic libraries arrayed in yeast shuttle vectors. The invention is also directed to methods of using targeting constructs made by the methods to generate transgenic animals.Type: GrantFiled: May 10, 2000Date of Patent: January 7, 2003Assignee: Amgen Inc.Inventor: Sushil K. Thukral
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Publication number: 20030003553Abstract: Disclosed are novel strains of Rhizopus oryzae and uses thereof. The strains of the invention are temperature-resistant and convert a carbon source to lactic acid at high temperatures.Type: ApplicationFiled: April 23, 2002Publication date: January 2, 2003Applicant: Archer-Daniels-Midland Company.Inventor: Hungming J. Liaw
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Patent number: 6500644Abstract: A method for in vivo production of a library in cells comprising a multitude of mutated genetic elements, wherein an error-prone polymerase is used in each ancestral cell to replicate all or a part of a genetic element independently of the host chromosomal replication machinery. The genetic element comprises i) an origin of replication from which replication is initiated, ii) optionally a genetic marker, e.g. a gene conferring resistance towards an antibiotic, iii) a gene encoding the polypeptide of interest. Also methods for the generation of a DNA sequence encoding a desired variant of a polypeptide of interest, and for the determination of such a DNA sequence are described.Type: GrantFiled: September 26, 2000Date of Patent: December 31, 2002Assignee: Novozymes A/SInventors: Torben Vedel Borchert, Stanislas Dusko Ehrlich
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Publication number: 20020197627Abstract: Disclosed are methods for identifying molecular interactions (e.g., protein/protein, protein/DNA, protein/RNA, or RNA/RNA interactions). All of the methods within the invention employ counterselection and at least two hybrid molecules. Molecules which interact reconstitute a transcription factor and direct expression of a reporter gene, the expression of which is then assayed. Also disclosed are genetic constructs which are useful in practicing the methods of the invention.Type: ApplicationFiled: March 27, 2002Publication date: December 26, 2002Applicant: The General Hospital Corporation, a Massachusetts corporationInventors: Marc Vidal, Jef D. Boeke, Ed Harlow
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Publication number: 20020197598Abstract: This invention provides methods of determining whether a compound inhibits HIV-1 reverse transcriptase. This invention provides methods of determining whether a compound inhibits formation of a complex between a p66 and p51 subunit polypeptides of HIV-1 reverse transcriptase. This invention provides a method of determining whether a compound enhances formation of a complex between a p66 and p51 subunit polypeptides of HIV-1 reverse transcriptase. This invention provides methods of determining whether a compound inhibits formation of a complex between two p66 subunit polypeptides of HIV-1 reverse transcriptase. This invention provides methods of determining whether a compound enhances formation of a complex between two p66 subunit polypeptides of HIV-1 reverse transcriptase.Type: ApplicationFiled: June 12, 2002Publication date: December 26, 2002Inventors: Stephen P. Goff, Gilda Tachedjian
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Publication number: 20020182709Abstract: 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 heteroulogous 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: ApplicationFiled: June 9, 2002Publication date: December 5, 2002Inventors: Dean Dawson, John Swindle
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Publication number: 20020160408Abstract: The present invention relates to a new yeast two-hybrid system for the detection of interactions between membrane proteins and lumenal proteins of the endoplasmic reticulum. The yeast two-hybrid system of the present invention uses the IRE1 gene of yeast which is a key element in the endoplasmic reticulum unfolded protein response to signal the interaction between proteins within the endoplasmic reticulum. The IRE1 gene codes for a type 1 membrane protein Ire1p, that has a kinase in the cytosolic domain and it signals the presence of unfolded proteins in the ER by oligomerization and transphosphorylation, this in turn activates a RNaseL that has as its unique (so far) substrate the HAC1 messenger RNA, that codes for the transcription factor that binds to the unfolded protein response element and upregulates transcription of ER protein required for folding proteins within the ER.Type: ApplicationFiled: April 10, 2002Publication date: October 31, 2002Applicant: NATIONAL RESEARCH COUNCIL OF CANADAInventors: Marc F. Pelletier, Gregor Jansen, John J.M. Bergeron, David Y. Thomas
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Publication number: 20020160380Abstract: The present invention relates to a method for producing combinatorial functional expression libraries using a combinatorial library of nucleic acids belonging to the same gene family, comprising a step of cloning by recombination in yeast. The invention also relates to a method for producing functional mosaic proteins and for analyzing a combinatorial functional expression library, by determining a sequential footprint for each of the mosaic proteins of the library.Type: ApplicationFiled: March 14, 2002Publication date: October 31, 2002Inventors: Gilles Truan, Valerie Abecassis, Denis Pompon
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Publication number: 20020155549Abstract: The present invention provides the promoter clone discovery of a glucoamylase gene of a starch utilizing yeast strain Schwanniomyces castellii. The isolated glucoamylase promoter is an inducible promoter, which can regulate strong gene expression in starch culture medium.Type: ApplicationFiled: August 2, 2001Publication date: October 24, 2002Inventors: Johnway Gao, Rodney S. Skeen, Brian S. Hooker, Daniel B. Anderson
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Publication number: 20020138880Abstract: A method of isolating genes conferring resistance to aluminium is provided and two particular aluminium tolerant genes are described. These genes are designated ALR1 and ALR2. The two tolerance genes were isolated from yeast strains but were found to have homology with bacterial genes responsible for divalent ion uptake. Hence a method of isolating divalent cation transporters is envisaged by using complementation of magnesium transporter mnutant strains of yeasts aluminium tolerance genes.Type: ApplicationFiled: January 12, 1998Publication date: September 26, 2002Inventors: RICHARD CLAGUE GARDNER, COLIN WHITI MACDIARMID, ROBERT JOHN HAY
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Patent number: 6451559Abstract: A new expression vector for the production of a polypeptide in yeast. The vector includes a sequence coding for the polypeptide and other sequences allowing expression of the polypeptide only in yeast. The other sequences lack any non-yeast sequences. Other embodiments include a yeast strain comprising such a vector, a method for the production of the vector, a method for the production of the yeast strain by transformation of a yeast strain with the new vector, and a method for the production of a polypeptide in the transformed yeast strain by fermentation thereof followed by isolation of the polypeptide.Type: GrantFiled: March 3, 2000Date of Patent: September 17, 2002Assignee: Pentapharm AGInventors: Thomas Schreier, Rainer Voegeli
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Publication number: 20020127724Abstract: The present invention provides the promoter clone discovery of an alpha-amylase gene of a starch utilizing yeast strain Schwanniomyces castellii. The isolated alpha-amylase promoter is an inducible promoter, which can regulate strong gene expression in starch culture medium.Type: ApplicationFiled: August 2, 2001Publication date: September 12, 2002Inventors: Johnway Gao, Rodney S. Skeen, Brian S. Hooker, Daniel B. Anderson
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Publication number: 20020123149Abstract: Yeast cells are mutagenized to obtain desirable mutants. Mutagenesis is mediated by a defective mismatch repair system which can be enhanced using conventional exogenously applied mutagens. Yeast cells with the defective mismatch repair system are hypermutable, but after selection of desired mutant yeast strains, they can be be rendered genetically stable by restoring the mismatch repair system to proper functionality.Type: ApplicationFiled: February 21, 2001Publication date: September 5, 2002Inventors: Nicholas C. Nicolaides, Philip M. Sass, Luigi Grasso, Bert Vogelstein, Kenneth W. Kinzler
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Publication number: 20020119439Abstract: A method for assaying trinucleotide repeat mutations in eukaryotic cells is provided.Type: ApplicationFiled: December 21, 2000Publication date: August 29, 2002Inventors: Robert S. Lahue, Juan J. Miret, Richard Pelletier
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Publication number: 20020115220Abstract: An object of the present invention is to provide a vector which can be integrated into a yeast chromosome in a high number of copies. Another object of the present invention is to provide a modified vector which can be integrated into the yeast chromosome in a high number of copies and of which expression units stably maintain on the chromosome. The vector according to the present invention comprises a marker gene for selecting transformants, a shortened promoter sequence which is operably linked to the marker gene and a sequence homologous to the chromosomal DNA of Candida utilis, and optionally a heterologous gene or a gene derived from C. utilis, wherein the vector is linearized by cleaving within said homologous DNA sequence or at both ends of the homologous DNA sequence with restriction enzymes, and wherein the heterologous gene or the gene derived from C. utilis can be integrated into the chromosomal DNA of C. utilis by homologous recombination.Type: ApplicationFiled: July 20, 2001Publication date: August 22, 2002Inventors: Keiji Kondo, Yutaka Miura
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Patent number: 6433137Abstract: The present invention provides TUP1 polynucleotides, including TUP1 polynucleotides encoding Tup1, and Tup1 polypeptides, from Candida albicans. Disruption of TUP1 function in C. albicans is associated with filamentous formation as well as low infectivity. These TUP1 polynucleotide and Tup1 polypeptide sequences (and anti-Tup1 antibodies derived from Tup1 polypeptides) may be used in methods of detecting C. albicans sequences in a biological sample. Further, the invention provides methods for screening agents which may control C. albicans virulence and compositions comprising these agents. The invention also provides methods of obtaining gene(s) and/or gene product(s) which are involved in a TUP1 pathway, as well as methods of controlling C. albicans virulence by comprising TUP1 function.Type: GrantFiled: July 1, 1998Date of Patent: August 13, 2002Assignee: The Regents of the University of CaliforniaInventors: Alexander D. Johnson, Burkhard Braun
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Publication number: 20020106698Abstract: A method for detecting protein-protein interactions is provided, in which two fusion proteins are prepared and allowed to interact with each other in yeast cells. The interaction between the two fusion proteins leads to protein trans-splicing, generating an active and detectable reporter.Type: ApplicationFiled: January 4, 2002Publication date: August 8, 2002Applicant: Myriad Genetics, IncorporatedInventor: John Manfredi
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Publication number: 20020106699Abstract: A method for detecting protein-protein interactions is provided, in which two fusion proteins are prepared and allowed to interact with each other. The interaction between the two fusion proteins leads to protein trans-splicing, generating an active and detectable reporter.Type: ApplicationFiled: January 4, 2002Publication date: August 8, 2002Inventors: John Manfredi, Jay Zhang
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Publication number: 20020103154Abstract: The present invention relates to methods of identifying genes in Saccharomyces cerevisiae which are essential for germination and proliferation of S. cerevisiae and using the identified genes or their encoded proteins as targets for highly specific antifungal agents, insecticides, herbicides and anti-proliferation drugs. The present invention also provides a method to systematically analyze the S. cerevisiae genome to identify essential genes for use as targets for antifungal agents, insecticides, herbicides and anti-proliferation drugs. The present invention provides antisense molecules and ribozymes comprising sequences complementary to the sequences of mRNAs of essential genes that function to inhibit the essential genes. The present invention also provides neutralizing antibodies to proteins encoded by essential genes that bind to and inactivate the essential gene products.Type: ApplicationFiled: September 27, 2001Publication date: August 1, 2002Applicant: Rosetta Inpharmatics, Inc.Inventor: Dago Dimster-Denk
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Patent number: 6410272Abstract: A method for the production and secretion of proteins with hirudin activity in an eukaryotic host organism is provided. There are also provided hybrid vectors comprising a DNA sequence encoding a signal peptide upstream of and in reading frame with the structural gene for desulphatohirudin, and eukaryotic host organisms transformed with said hybrid vectors.Type: GrantFiled: February 27, 1990Date of Patent: June 25, 2002Assignee: Ciba-Geigy Corporation a Corp. of New YorkInventors: Bernd Meyhack, Walter Märki, Jutta Heim
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Patent number: 6410271Abstract: Methods are provided for generating highly diverse libraries of expression vectors encoding fusion proteins such as single-chain antibodies via homologous recombination in yeast. The method comprises: transforming into yeast cells a linearized yeast expression vector having a 5′- and 3′-terminus sequence at the site of linearization and a library of insert nucleotide sequences that are linear and double-stranded; and having homologous recombination occur between the vector and the insert sequence such that the insert sequence is included in the vector in the transformed yeast cells.Type: GrantFiled: June 23, 2000Date of Patent: June 25, 2002Assignee: GeneTasTix CorporationInventors: Li Zhu, Shaobing Benjamin Hua
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Publication number: 20020068325Abstract: The invention provides methods and compositions for the highly efficient production of heterologous proteins in yeast and other fungi by overcoming the previous problems associated with failure of these proteins to fold properly. According to the invention, the quality control mechanism employed by fungi which returns misfolded proteins to the cytosol for degradation is manipulated so that these proteins are instead secreted.Type: ApplicationFiled: December 5, 2001Publication date: June 6, 2002Inventors: Davis T.W. Ng, Shilpa Vashist
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Patent number: 6391642Abstract: The invention is directed to a method of making a yeast artificial chromosome (YAC) comprising introducing into yeast cells a population of nucleic acids and a vector, wherein the vector comprises a yeast centromere, a selectable marker, a yeast telomere, and a sequence which can recombine with a region of a nucleic acid within the population of nucleic acids, whereby in vivo recombination makes the YAC. The invention is also directed to a method of making a YAC using two vectors and a method of making a circular YAC. The invention is also directed toward methods of making YACs with a selected nucleic acid insert from a mixed population of nucleic acids using transformation-associated recombination.Type: GrantFiled: April 14, 1998Date of Patent: May 21, 2002Assignee: The United States of America as represented by the Department of Health and Human ServicesInventors: Michael A. Resnick, Vladimir L. Larionov, Natalay Y. Kouprina, Edward L. Perkins
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Patent number: 6391582Abstract: 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: December 9, 1998Date of Patent: May 21, 2002Assignee: Rigel Pharmaceuticlas, Inc.Inventors: Ying Luo, Pei Wen Yu, James Lorens
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Patent number: 6379924Abstract: The use of a means to vary Ubc4p or Ubc5p activity in a fungal cell to control the copy number of a plasmid in the cell. The level of Ubc4p or Ubc5p activity may be reduced/abolished (for example by gene deletion, mutagenesis to provide a less active protein, production of antisense mRNA or production of competitive peptides) to raise the copy number and increase yield of a protein encoded by the plasmid.Type: GrantFiled: December 15, 1999Date of Patent: April 30, 2002Assignee: Delta Biotechnology Ltd.Inventor: Darrell Sleep
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Patent number: 6380465Abstract: The present invention provides, inter alia, nucleic acids which encode P450s in corn that, when expressed in the presence of a reductase, metabolize compounds exemplary of several distinct classes of insecticides and herbicides. The invention also includes amino acids encoded by the nucleic acids, as well as vectors, cells and eukaryotes comprising the nucleic or amino acid compounds. Also included are methods using the materials provided.Type: GrantFiled: July 11, 1999Date of Patent: April 30, 2002Assignee: University of Kentucky Research FoundationInventor: Michael Barrett
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Publication number: 20020045233Abstract: The invention provides cloned ketoreductase genes, vectors for expressing same, recombinant host cells that express said vector-borne genes, a method for stereospecifically reducing a ketone using a recombinant ketoreductase, or a recombinant host cell that expresses a cloned ketoreductase gene, as well as a method for generating polynucleotide sequences more conducive to recombinant expression.Type: ApplicationFiled: October 8, 1999Publication date: April 18, 2002Inventors: CHARLES LEE HERSHBERGER, ROBERT ALLEN PAYSON
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Patent number: 6365409Abstract: 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: September 23, 1999Date of Patent: April 2, 2002Assignee: Anadys Pharmaceuticals, IncInventors: John D. Bradley, Craig M. Thompson, Jeffrey B. Moore, C. Richard Wobbe, Judith M. Healy, Caroline E. Donnelly
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Patent number: 6361969Abstract: An expression system which provides heterologous proteins expressed by a non-native host organism but which have native-protein-like biological activity and/or structure. Disclosed are vectors, expression hosts and methods for expressing the heterologous proteins. The expression system involves co-expression of protein factor(s) which is/are capable of catalyzing disulphide bond formation and desired heterologous protein(s). The expression system is presented using yeast cells as the preferred host, protein disulphide isomerase (PDI) and thioredoxin (TRX) as the preferred examples of the protein factors and HCV-E2715 envelope glycoprotein and human FIGF as the preferred examples of the heterologous proteins.Type: GrantFiled: August 16, 1999Date of Patent: March 26, 2002Assignee: Chiron S.p.A.Inventor: Cesira Galeotti
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Publication number: 20020034788Abstract: A nucleic acid sequence including a CYP promoter operably linked to nucleic acid encoding a heterologous protein is provided to increase transcription of the nucleic acid. Expression vectors and host cells containing the nucleic acid sequence are also provided. The methods and compositions described herein are especially useful in the production of polycarboxylic acids by yeast cells.Type: ApplicationFiled: July 24, 2001Publication date: March 21, 2002Inventors: David L. Craft, C. Ron Wilson, Dudley Eirich, Yeyan Zhang
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Patent number: 6358705Abstract: The present invention relates to a method for expressing heterologous proteins or polypeptides in yeast by culturing a transformed yeast strain which does not contain a functional antibiotic resistance marker gene.Type: GrantFiled: July 6, 1999Date of Patent: March 19, 2002Assignee: Novo Nordisk A/SInventors: Thomas Kjeldsen, Knud Vad
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Patent number: 6342375Abstract: In the present system an adequate expression system for the production and secretion of biologically active human growth hormone (HGH) in its natural form in which a methylotrophic yeast such as Pichia pastoris is used as host organism has been developed. This invention includes a methylotrophic yeast transformed with at least one copy of a functional cDNA sequence encoding HGH, which is functionally associated with a second DNA sequence encoding the S. cerevisae alpha factor pre-pro sequence (including the proteolytic processing site: lys-arg), and in which both DNA sequences are under the regulation of a methylotrophic yeast gene promoter which is inducible with methanol. Methylotrophic yeasts containing in their genome at least one copy of the DNA sequence efficiently produce and secrete mature, correctly processes and biologically active HGH, into the culture medium.Type: GrantFiled: July 21, 1999Date of Patent: January 29, 2002Assignee: Universidad Autonoma de Nuevo LeonInventors: Martha Guerrero Olazaran, Hugo Barrera Saldana, Jose Maria Viader Salvado