Abstract: Provided herein are mutant DNA-dependent polymerases which are derived from, or otherwise related to, wild type RB69 DNA polymerase. These mutant polymerases are capable of selectively binding labeled nucleotides. These mutant polymerases are also capable of incorporating a variety of naturally occurring and modified nucleotides, including, for example, terminator nucleotides.
Type:
Grant
Filed:
July 1, 2016
Date of Patent:
March 14, 2017
Assignee:
Life Technologies Corporation
Inventors:
Marian Peris, Michael Phelan, Barnett Rosenblum, Stephen Hendricks
Abstract: The invention provides nucleic acids and polypeptides for a nucleic acid polymerase from a thermophilic organism, Thermus scotoductus. The invention also provides methods for using these nucleic acids and polypeptides.
Abstract: Fusion proteins comprising a single strand DNA binding protein and a nucleic acid polymerase (e.g. DNA polymerase or reverse transcriptase). These high fidelity proteins are suitable for use in nucleic acid amplification methods, including the polymerase chain reaction (PCR).
Type:
Grant
Filed:
August 4, 2014
Date of Patent:
February 28, 2017
Assignee:
LIFE TECHNOLOGIES CORPORATION
Inventors:
Jun Lee, Robert Potter, David Mandelman
Abstract: A recombinant polypeptide is described including at least one PUF RNA-binding domain capable of specifically binding to a cytosine RNA base. The PUF RNA-binding domain of the polypeptide includes at least one RNA base-binding motif of the general formula X1X2X3X4X5X6X7X8X9X10X11 wherein X1 is selected from a defined group and wherein the RNA base-binding motif is operably capable of specifically binding to a cytosine RNA base.
Abstract: Disclosed herein are conjugates comprising a biomolecule linked to a label that have biological activity and are useful in a wide variety of biological applications. For example, provided herein are labeled polymerase conjugates including a polymerase linked to one or more labels, wherein the conjugate has polymerase activity. Such conjugates can exhibit enhanced biological activity and/or superior detectability as compared to conventional labeled polymerases. Also disclosed herein are improved methods for preparing such conjugates, and methods and systems for using such conjugates in biological applications such as nucleotide incorporation, primer extension and single molecule sequencing.
Abstract: Compositions that include polymerases with features for improving entry of nucleotide analogues into active site regions and for coordinating with the nucleotide analogues in the active site region are provided. Methods of making the polymerases and of using the polymerases in sequencing and DNA replication and amplification as well as kinetic models of polymerase activity and computer-implemented methods of using the models are also provided.
Type:
Grant
Filed:
November 11, 2014
Date of Patent:
January 31, 2017
Assignee:
Pacific Biosciences of California, Inc.
Inventors:
David K. Hanzel, Geoff Otto, Paul Peluso, Thang Pham, David R. Rank, Paul Mitsis, Fred Christians, Arekadiusz Bibillo, Insil Park, Sonya Clark, John Lyle
Abstract: The present invention pertains to a mutated T7 RNA polymerase and its use, the T7 RNA polymerase being mutated at position 744, the glutamine (Q) being replaced by an amino acid selected from arginine (Q744R), leucine (Q744L) or proline (Q744P).
Type:
Grant
Filed:
October 8, 2012
Date of Patent:
January 10, 2017
Assignees:
bioMérieux, Commissariat à l'Energie Atomique et Aux Energies Alternatives
Inventors:
Jean-Claude Boulain, Janie Dassa, Frédéric Ducancel, Bruno H. Muller, Alain Troesch, Laurent Mesta
Abstract: The present invention provides mutants of DNA polymerases having an increased rate of incorporation of nucleotides into nucleic acids undergoing polymerization and having an enhanced resistance to inhibitors of DNA polymerase activity. The mutant polymerases are well suited for fast PCR applications, for PCR amplification of targets in samples that contain inhibitors of wild-type polymerases, and for fast PCR amplification of samples containing DNA polymerase inhibitors. In exemplary embodiments, the mutants are mutants of Taq DNA polymerase.
Type:
Grant
Filed:
August 2, 2010
Date of Patent:
December 20, 2016
Assignee:
Agilent Technologies Inc.
Inventors:
Holly Hogrefe, Michelle Cayouette, Jeffrey Fox, Connie J. Hansen, Jennifer Lapira, Bahram Arezi, Nancy McKinney
Abstract: The present invention relates to variant endoglucanases having improved thermoactivity, improved thermostability, and improved viscosity reduction activity over wild-type M. thermophila endoglucanase.
Type:
Grant
Filed:
November 14, 2014
Date of Patent:
December 6, 2016
Assignee:
CODEXIS, INC.
Inventors:
Xiyun Zhang, Sachin Patil, Jie Yang, Ish Kumar Dhawan
Abstract: The present invention provides multi-component inhibitors of nucleic acid polymerases, methods of making, and methods of using same. One component of the multi-component inhibitor is a molecule that binds to a polymerase (i.e., a polymerase-binding molecule (PBM)), but does not thereby substantially inhibit its polymerase activity. Another component is a molecule or complex of molecules that binds to a PBM (i.e., a PBM-binding molecule). The combination of the PBM and PBM-binding molecule/complex substantially inhibits polymerase activity. The disclosed multi-component inhibitors are useful for DNA sequencing, nucleic acid amplification, cloning and synthesis, and the like.
Abstract: There is presently provided a probe comprising an isolated sphingolipid binding domain (SBD) polypeptide, wherein the isolated SBD polypeptide is capable of binding to a sphingolipid, and methods and uses relating to such a probe.
Type:
Grant
Filed:
September 12, 2007
Date of Patent:
November 22, 2016
Assignee:
Agency for Science, Technology and Research
Abstract: Methods and compositions are provided wherein microorganisms are used to alter the interface of hydrocarbons and hydrocarbon-coated surfaces to increase oil recovery, for improved bioremediation and/or to benefit pipeline maintenance.
Type:
Grant
Filed:
November 4, 2014
Date of Patent:
November 22, 2016
Assignee:
E I DU PONT DE NEMOURS AND COMPANY
Inventors:
Edwin R Hendrickson, Abigail K Luckring, Michael P Perry
Abstract: This invention provides for an improved generation of novel nucleic acid modifying enzymes. The improvement is the fusion of a sequence-non-specific nucleic-acid-binding domain to the enzyme in a manner that enhances the ability of the enzyme to bind and catalytically modify the nucleic acid.
Abstract: The invention relates to modified polymerase enzymes which exhibit improved incorporation of nucleotide analogs bearing substituents at the 3? position of the sugar moiety that are larger in size than the naturally occurring 3? hydroxyl group. Also described are methods of using the polymerases to incorporate nucleotides into polynucleotides, particularly in the context of DNA sequencing.
Type:
Grant
Filed:
September 3, 2014
Date of Patent:
September 20, 2016
Assignee:
Illumina Cambridge Limited
Inventors:
Geoffrey Paul Smith, David Mark Dunstan Bailey, Raquel Maria Sanches-Kuiper, Harold Swerdlow, David James Earnshaw
Abstract: Disclosed are rationally-designed, non-naturally-occurring meganucleases in which a pair of enzyme subunits having specificity for different recognition sequence half-sites are joined into a single polypeptide to form a functional heterodimer with a non-palindromic recognition sequence. The invention also relates to methods of producing such meganucleases, and methods of producing recombinant nucleic acids and organisms using such meganucleases.
Abstract: This patent applications concerns compositions of matter that are DNA polymerases, where those polymerases have had one or more of their amino acids replaced at sites chosen by an analysis of patterns of conservation and replacement within homologous protein sequences. Disclosed here are sites within Family A DNA polymerases where amino acid replacement creates polymerases having utility, in an example where DNA nucleotides are incorporated having modified or unnatural nucleobases, and/or nucleotides whose sugar is unnatural or derivatized, including 3?-O-amino-2?-deoxyribonucleoside triphosphates. The claimed compositions include polymerases that hold amino acid replacements at claimed sites in Taq polymerase, and are prepared by site-directed mutagenesis that modifies a gene encoding a parent Taq gene (natural or already mutated) to change the codon encoding the amino acid at the claimed site, giving a variant gene that encodes a Taq polymerase protein that has a different amino acid at the claimed site.
Abstract: The present invention provides cold shock protein-containing compositions for improved DNA synthesis reactions with improved reactivity, methods for synthesizing DNA using such compositions, kits for use in such methods, and DNA compositions yielded by such methods. The present invention further provides cold shock protein-containing compositions for the identification of endoribonuclease cleavage sites, methods for identifying endoribonuclease cleavage sites using such compositions, and kits for use in such methods.
Type:
Grant
Filed:
March 2, 2009
Date of Patent:
August 23, 2016
Assignee:
Rutgers, The State University of New Jersey
Abstract: Disclosed are mutant DNA polymerases having increased 3?-mismatch discrimination relative to a corresponding, unmodified polymerase. The mutant polymerases are useful in a variety of disclosed primer extension methods. Also disclosed are related compositions, including recombinant nucleic acids, vectors, and host cells, which are useful, e.g., for production of the mutant DNA polymerases.
Type:
Grant
Filed:
April 16, 2014
Date of Patent:
August 9, 2016
Assignee:
Roche Molecular Systems, Inc.
Inventors:
Fred Reichert, Keith Bauer, Thomas W. Myers, Nancy J. Schoenbrunner, Joseph San Filippo
Abstract: Provided are compositions comprising recombinant DNA polymerases that include amino acid substitutions, insertions, deletions, and/or exogenous features that confer modified properties upon the polymerase for enhanced single molecule sequencing or nucleic acid amplification. Such properties include enhanced performance with large nucleotide analogs, increased stability, increased readlength, and improved detection of modified bases, and can also include resistance to photodamage, enhanced metal ion coordination, reduced exonuclease activity, reduced reaction rates at one or more steps of the polymerase kinetic cycle, decreased branching fraction, altered cofactor selectivity, increased yield, increased accuracy, altered speed, increased cosolvent resistance, and the like. Also provided are nucleic acids which encode the polymerases with the aforementioned phenotypes, as well as methods of using such polymerases to make a DNA or to sequence a DNA template.