Abstract: A method of producing a protein of interest (POI) by culturing a recombinant eukaryotic cell line comprising an expression construct comprising a regulatable promoter and a nucleic acid molecule encoding a POI under the transcriptional control of said promoter, comprising the steps a) cultivating the cell line with a basal carbon source repressing the promoter, b) cultivating the cell line with a limited amount of a supplemental carbon source de-repressing the promoter to induce production of the POI at a transcription rate of at least 15% as compared to the native pGAP promoter, and c) producing and recovering the POI; and further an isolated regulatable promoter and a respective expression system.
Type:
Grant
Filed:
October 5, 2012
Date of Patent:
December 6, 2016
Assignee:
Lonza Ltd.
Inventors:
Diethard Mattanovich, Brigitte Gasser, Michael Maurer, Roland Prielhofer, Joachim Klein, Jana Wenger
Abstract: Compositions and methods for nitrogen sensitive regulation of expression of a transcribable nucleic acid molecule. One aspect provides a nitrogen-sensitive expression system that includes a transcription factor region comprising an NtcA binding site and a core promoter region comprising a RuBisCo promoter or a variant or a functional fragment thereof. Another aspect provides a method of transforming a host cell with an expression system. Also provided are expression cassettes, transformed host cells, and kits.
Type:
Grant
Filed:
March 26, 2012
Date of Patent:
November 29, 2016
Assignee:
Proterro, Inc.
Inventors:
Robert J. Turner, Valerie Sershon, John Aikens, Denise Holzle
Abstract: Disclosed are isolated nucleic acid molecules that have promoter activity specific to xylose. The synthetic promoters, SEQ ID NO: 2, SEQ ID NO: 3, and SEQ ID NO: 4, promote the expression of a coding region of interest in transformed yeast cells.
Type:
Grant
Filed:
July 10, 2015
Date of Patent:
November 29, 2016
Assignee:
The United States of America, as represented by the Secretary of Agriculture
Abstract: The present invention relates to methods for constructing a recombinant fungal host cell comprising one or more copies of a polynucleotide construct integrated in its genome, said method comprising transforming a fungal host cell with an integrative polynucleotide construct comprising a first polynucleotide encoding a selectable marker, wherein the first polynucleotide, a 5? untranslated region thereof and/or a riboswitch operably linked therewith comprises a spliceosomal intron which has 5 nucleotides or less between its branch site and its acceptor site; and a second polynucleotide encoding a polypeptide of interest; as well as suitable polynucleotide constructs, resulting fungal host cells and methods of manufacture.
Abstract: The invention relates to the fields of screening assays, compounds, and methods for altering gene expression and protein levels. In particular, the invention includes assays to screen for agents capable of modulating gene expression in a UTR-dependent manner and agents capable of modulating gene expression.
Type:
Grant
Filed:
May 22, 2015
Date of Patent:
October 25, 2016
Assignee:
PTC THERAPEUTICS, INC.
Inventors:
Liangxian Cao, Nikolai Naryshkin, Charles Romfo, Matthew C. Pellegrini, Anuradha Roy, Panayiota Trifillis, Christopher R. Trotta
Abstract: An object to be solved by the present invention is to provide, for example, a method for producing a Kluyveromyces marxianus transformant by a method for conveniently and efficiently connecting the ends of DNA fragments without using specific restriction enzymes or their recognition sequences, and a method for producing a useful substance using the transformant.
Type:
Grant
Filed:
February 1, 2011
Date of Patent:
September 27, 2016
Assignee:
Yamaguchi University
Inventors:
Rinji Akada, Hisashi Hoshida, Babiker Mohamed Ahmed Abdel-Banat, Jun Asakawa
Abstract: According to one embodiment, a reporter vector presenting an extracellular binding capacity to metallic compounds contains a nucleotide sequence exhibiting a promoter activity depending on a specific condition, a nucleotide sequence encoding a metallic compound-binding peptide presented extracellularly, and a nucleotide sequence encoding transcription termination signals.
Abstract: The invention encompasses method of using quantitative PCR to detect fungal organisms in clinical and environmental samples to generate standards that allow the quantification of fungal organisms in the samples.
Type:
Grant
Filed:
May 12, 2014
Date of Patent:
September 6, 2016
Assignees:
THE TRANSLATIONAL GENOMICS RESEARCH INSTITUTE, ARIZONA BOARD OF REGENTS ON BEHALF OF NORTHERN ARIZONA UNIVERSITY
Inventors:
Cindy Liu, Sergey Kachur, Lance Price, Paul Keim
Abstract: A polynucleotide comprising a first region the 5? end of which is complementary to a portion of a target nucleic acid, a cleavable second region, a third region having a stem-loop structure, and a fourth region complementary to the 3? end of the first region, and use of the polynucleotide, as well as a composition comprising two such polynucleotides each of which hybridize different strands of a double-stranded target nucleic acid, and methods and kits using the same for amplifying targets.
Type:
Grant
Filed:
May 9, 2014
Date of Patent:
August 30, 2016
Assignee:
SAMSUNG ELECTRONICS CO., LTD
Inventors:
Sea-hee Kim, Joon-ho Kim, Joo-won Rhee, Sun-ok Jung, Ko-bong Choi, Kyu-youn Hwang
Abstract: The present invention relates to a method for producing natively folded disulfide bond containing proteins in a prokaryotic host. The method comprises that in the cytoplasm of a prokaryotic cell is expressed protein(s) of interest that naturally contain disulfide bonds and naturally occurring or inverted transmembrane enzyme, wherein the cysteines of the active site(s) are naturally or after genetic engineering located towards the prokaryotic cytoplasm. The enzyme is selected from the group of VKOR, inverted VKOR (iVKOR) and inverted Dsb B (iDsb B). In the prokaryotic cell is also expressed cytoplasmic DsbA or a corresponding protein being capable of providing electrons to the active site(s) of VKOR, iVKOR or iDsbB. The invention relates also to a prokaryotic host cell and a vector system for producing natively folded disulfide bond containing proteins.
Abstract: Disclosed herein are systems and methods for producing recombinant proteins utilizing mutant E. coli strains containing expression vectors carrying nucleic acids encoding the proteins, and secretory signal sequences to direct the secretion of the proteins to the culture medium. Host cells transformed with the expression vectors are also provided.
Type:
Grant
Filed:
July 27, 2011
Date of Patent:
August 9, 2016
Assignees:
WISCONSIN ALUMNI RESEARCH FOUNDATION, CORNELL UNIVERSITY
Inventors:
Sydnor T. Withers, III, Miguel A. Dominguez, Matthew P. DeLisa, Charles H. Haitjema
Abstract: The present invention concerns a method of producing a desired heterologous gene product wherein said heterologous gene product is expressed from a strong promoter, said method comprising expressing said gene using a mutant mRNA leader which comprises one or more mutations which enhance transcription of the gene. The invention also provides a mutant Pm mRNA leader sequence, and a vector and a library comprising the leader sequence. Methods of obtaining an mRNA mutant leader and identifying an mRNA mutant leader are encompassed, along with a vector for selection or identification of an mRNA leader mutant and a use thereof for screening.
Type:
Grant
Filed:
August 3, 2007
Date of Patent:
July 12, 2016
Assignee:
VECTRON BIOSOLUTIONS AS
Inventors:
Svein Valla, Rahmi Lale, Laila Berg, Ingrid Bakke
Abstract: The present disclosure provides yeast transformed with a nucleic acid molecule (GAT1) to reduce nitrogen catabolite repression of asparagine transport/degradation and/or overexpress genes (ASP1 or ASP3) encoding cell-wall or extracellular proteins involved in asparagine degradation and/or genes (AGP1 or GNP1 or GAP1) encoding proteins involved in asparagine transport under food preparation/processing conditions. The genetically modified yeast has enhanced ability to reduce acnlamide concentration in foods prepared by heating. Also provided are methods and uses of the transgenic yeast for reducing acnlamide in a food product and food products having reduced acrylamide content prepared using the transgenic yeast.
Abstract: Reduced genome bacteria with improved genetic stability are provided. Also provided are methods of producing polypeptides using the reduced genome bacteria with improved genetic stability.
Type:
Grant
Filed:
October 19, 2012
Date of Patent:
May 17, 2016
Assignee:
SCARAB GENOMICS, LLC
Inventors:
Frederick R. Blattner, Balint Csorgo, Gyorgy Posfai
Abstract: The present invention relates to novel engineered lower eukaryotic host cells for expressing heterologous proteins and to methods of generating such strains.
Type:
Grant
Filed:
October 23, 2012
Date of Patent:
May 3, 2016
Assignee:
Merck Sharp & Dohme Corp.
Inventors:
Bo Jiang, Rebecca D. Argyros, Stephanie Nelson, Robert C. Davidson, Ronghua Chen, Jun Zhuang