Abstract: The present invention provides a method of producing a cloned insert that is representative of the ends of a large segment of DNA from the genome of an organism. Specifically, the present invention provides a clone insert strategy that comprising the steps of: a) isolating a nucleic acid molecule, or portion thereof, wherein said nucleic acid molecule is at least 20 kb; b) ligating said nucleic acid molecule into a vector molecule; c) cutting said nucleic acid molecule with a restriction endonuclease, wherein said restriction endonuclease cuts said isolated nucleic acid molecule of step (a) in at least two sites but does not cut said vector DNA, d) ligating said cut nucleic acid molecule; e) transforming a host cell with said ligated nucleic acid molecule to propagate said molecule; f) determining the sequence of the ends of the nucleic acid insert and the sequence from the restriction endonculease site.
Type:
Grant
Filed:
September 16, 2002
Date of Patent:
September 13, 2005
Assignee:
Applera Corporation
Inventors:
Robert Holt, Hamilton O. Smith, Cynthia Pfannkoch
Abstract: This invention is directed to a regulatory region obtained from a wheat aleurone gene LtpW1. This regulatory region, truncated derivatives, mutations, or deletions of this regulatory region, can be used to express heterologous genes of interest within aleurone cells of a plant. Furthermore, this invention is directed to a truncated LtpW1 regulatory region that exhibits constitutive activity with both monocot and dicot plants. This invention is also directed to vectors comprising these regulatory regions operatively linked with a heterologous gene of interest, as well as plant cell cultures and transgenic plants comprising these vectors. A method for the preparation of a plant using the regulatory regions of this invention are also disclosed.
Type:
Grant
Filed:
October 13, 2000
Date of Patent:
September 6, 2005
Assignee:
Her Majesty the Queen in Right of Canada, as represented by the Minister of Agriculture and Agri-Food
Inventors:
John Simmonds, Leslie Cass, Linda Harris, Sharon Allard, Kamal Malik, Teresa Martin-Heller, Dan Brown, Ming Hu, Brian Miki, Keqiang Wu
Abstract: A method for cloning a nucleic acid fragment into a vector by flanking the fragment with first and second adapter sequences, and contacting the fragment with the vector having sequences homologous to the first and second adapter sequences under conditions such that the nucleic acid fragment is incorporated into the vector by homologous recombination in vivo in a host cell. Additionally, a method for selecting for a successful transformation of a vector by a nucleic acid insert. Also, systems for cloning a nucleic acid fragment into a vector without at least one of a restriction enzyme, a ligase, a gyrase, a single stranded DNA binding protein, or other DNA modifying enzymes. Further, a kit for cloning a nucleic acid fragment into a vector.
Type:
Grant
Filed:
April 16, 2002
Date of Patent:
August 30, 2005
Assignee:
Gene Therapy Systems, Inc.
Inventors:
Xiaowu Liang, Andy Teng, Shizhong Chen, Dongyuan Xia, Philip L. Felgner
Abstract: The isolation of and methods of using a sub-genomic transcript (Sgt) promoter from Mirabilis mosaic virus (MMV) are described. A 333 bp MMV Sgt promoter fragment (sequence?306 to +27 from the transcription start site, TSS) was found to be sufficient for strongest promoter activity. This MMV Sgt promoter fragment shows comparable promoter activity to the MMV FLt promoter both in transgenic plants and in protoplasts. The MMV Sgt promoter also demonstrates much greater activity compared to Cauliflower mosaic virus (CaMV) 19S promoter and 35S promoter. The MMV Sgt promoter fragment and any chimeric gene to which it may be linked are usefull for plant geneic engineering to obtain transgenic plants, plant cells and seeds.
Type:
Grant
Filed:
June 5, 2002
Date of Patent:
August 16, 2005
Assignee:
University of Kentucky Research Foundation
Abstract: The present invention is directed to a polynucleotide comprising open reading frames defining a coding region encoding a retinoblastoma binding protein (RBP-7) as well as regulatory regions located both at the 5? end and the 3? end of said coding region. The present invention also pertains to a polynucleotide carrying the natural regulation signals of the RBP-7 gene which is useful in order to express a heterologous nucleic acid in host cells or host organisms as well as functionally active regulatory polynucleotides derived from said regulatory region. The invention also concerns polypeptides encoded by the coding region of the RBP-7 gene. The invention also deals with antibodies directed specifically against such polypeptides that are useful as diagnostic reagents.
Abstract: The increased use of nucleotide sequence data mining techniques has amplified the demand for efficient methods of producing recombinant proteins in eukaryotic cells. A strategy is provided for enhancing the synthesis of recombinant amino acid sequences by polymerizing expression cassettes in vitro before producing recombinant hosts.
Abstract: The invention provides a synthesis-inducible eucaryotic promoter having a transcriptional activity comprising, for example, a heavy metal-sensitive DNA sequence, derived from a natural promotor, of a base sequence of any one of SEQ ID NOS: 1 to 4; an inducer of the transcriptional activation activity thereof; a recombinant expression vector comprising a site for the insertion of an extraneous gene ligated to the above promotor: a eucaryotic expression kit comprising eucaryotic cells transfected with the above vector and the above inducer; and a method for expression of an extraneous gene in a eucaryotic system by using the same
Abstract: Accessory functions capable of supporting efficient recombinant AAV (rAAV) virion production in a suitable host cell are provided. The accessory functions are in the form of one or more vectors that are capable of being transferred between cells. Methods of producing rAAV virions are also provided. The methods can be practiced to produce commercially significant levels of rAAV particles without also generating significant levels of infectious helper virus or other contaminating by-products.
Abstract: This invention relates to the field of gene expression. In particular this invention relates to the use of heterologous phytochromes to translocate polypeptides into the nucleus of a cell. Where the polypeptides comprise transactivators or repressors this invention provides a system for light-directed gene expression.
Type:
Grant
Filed:
May 29, 2002
Date of Patent:
May 3, 2005
Assignee:
The Regents of the University of California
Inventors:
John Clark Lagarias, Takayuki Kochi, Nicole Frankenberg, Gregory A. Gambetta, Beronda L. Montgomery
Abstract: The present invention provides novel myeloid cell specific promoters, and cis-acting elements that influence the activity of a myeloid cell specific promoters, as well as promoter-heterologous gene constructs and transfected myeloid cells which include the novel promoters and/or the novel cis-acting elements. The present invention also provides a method of transfecting myeloid cells and a method of producing a selected product within the transfected cells. The invention also includes a method for identifying factors that can regulate myeloid cell specific transcription.
Abstract: Under the regulation of a methanol-inducible alcohol oxidase gene (AOX1) promoter and a transcription termination sequence originating from Pichia pastoris, Pichia yeast, which has been transformed with an expression vector containing an MK family protein gene ligated to an a1 factor signal sequence originating from Saccharomyces cerevisiae, is cultured and an intact MK family protein is thus expressed and secreted in a large amount into the culture supernatant.
Abstract: The invention encompasses Drosophila Recombination Associated Protein (DRAP) isolated from D. melanogaster and a nucleic acid sequence encoding DRAP. The Drosophila Recombination Associated Protein, its homologues from other organisms or active peptides derived therefrom, as well as DNA encoding such protein are useful for homology-dependent pairing of three DNA strands. The combination of strand-transfer and topoisomerase activities associated with DRAP permits directed pairing and cleavage at defined site(s) within DNA. This in turn makes possible the isolation and/or removal of a defined segment of DNA. DRAP is also useful in cloning, genomic cloning and gene mapping, in promoting gene disruptions or “knockout” mutations, in carrying out targeted mutagenesis of specific genes and in generating transgenic animals.
Type:
Grant
Filed:
January 28, 2003
Date of Patent:
February 22, 2005
Assignee:
Albert Einstein College of Medicine of Yeshiva University
Abstract: The invention provides methods and compositions relating to novel human cellular inhibitor of apoptosis proteins (c-IAP1/2) comprising a series of defined structural domain repeats and/or a RING finger domain; in particular, at least two of: a particular first domain repeat, a particular second domain repeat, and a particular third domain repeat, and/or a particular RING finger domain. The proteins provide a c-IAP specific function, with preferred proteins being capable of modulating the induction of apoptosis; for example, by binding a human tumor necrosis factor receptor associated factor (TRAF). The compositions include nucleic acids which encode the subject c-IAP and hybridization probes and primers capable of hybridizing with the disclosed c-IAP genes. The invention includes methods of using the subject compositions in therapy, in diagnosis and in the biopharmaceutical industry.
Abstract: The present invention relates to intracellular receptors, and methods for the modulation of transcription using same. More particularly, the invention relates to the Nur family of nuclear receptors. In general aspects, the present invention relates to the idenfication of a physiologically relevant response element (RE) for Nur family members, an ER-10 element as well as to the idenfication of the type of protein-protein interactions of Nur family member, enabling their specific interaction with this RE-10 and tfeir modulation of transcription at physiologically relevant sites. The invention further relates to methods for modulating processes mediated by such nuclear receptors.
Type:
Grant
Filed:
December 12, 1997
Date of Patent:
February 8, 2005
Assignee:
Institut de Recherches Cliniques de Montreal
Inventors:
Jacques Drouin, Alexandra Philips, Mario Maira
Abstract: A process for selecting human cells for the production of human proteins by endogenous gene activation allows human proteins to be produced in economically feasible quantities and in a form suitable for producing a pharmaceutical composition. Also disclosed is a process for producing human proteins in a cell line identified in this matter.
Type:
Grant
Filed:
April 2, 2002
Date of Patent:
January 25, 2005
Assignee:
Roche Diagnostics GmbH
Inventors:
Michael Brandt, Reinhard Franze, Ulrich Pessara
Abstract: Described herein are methods for the enhanced production of secreted proteins. The secretion of a protein of interest having a substantially non-polar carboxy tail is enhanced by the placement of charged amino acid residues at the carboxy terminus either by adding to the native peptide or by replacing, i.e., substituting, the terminal residues of the native peptide.
Abstract: The invention provides primary and secondary cells that are transfected with a nucleic acid molecule that encodes erythropoietin, clonal or heterogenous strains of such cells, and methods of producing these cell strains.
Type:
Grant
Filed:
June 8, 1999
Date of Patent:
January 25, 2005
Assignee:
Transkaryotic Therapies, Inc.
Inventors:
Richard F. Selden, Douglas Treco, Michael W. Heartlein
Abstract: The invention concerns a novel construct for expressing a gene coding for a recombinant protein of interest placed under the control of the Ptrp tryptophan operon promoter in a procaryotic host cell. The invention is characterised in that the construct comprises a nucleic sequence capable of inactivating the gene coding for a TnaA tryptophanase when said nucleic sequence is introduced in said host cell. The invention also concerns vectors containing said construct and host cells transformed by said vectors. The invention further concerns methods for producing said recombinant proteins using said novel constructs.
Abstract: The invention relates to a genetic unit, optionally present as a multiple-copy, for inhibiting RNA. The unit contains the transcription units necessary for transcription by polymerase III and a DNA coding for inhibiting RNA, which is arranged within the unit in such a way that the transcribed RNA is part of the polymerase III transcript. Using these units it is possible to achieve increased stability of the inhibiting RNA, which may occur in the form of ribozymes or antisense-RNAs, whilst maintaining an undiminished activity.
The invention further relates to a process for introducing the genetic units into the cell, the use of these units and pharmaceutical preparations containing them.
Type:
Grant
Filed:
September 21, 1992
Date of Patent:
December 14, 2004
Assignee:
Boehringer Inglheim International GmbH
Inventors:
Hartmut Beug, Max L. Birnstiel, Matthew Cotten, Ernst Wagner, Harald Kandolf