专利号:US-7041479-B2 优先权日:2000-09-06 标题 :Enhanced in vitro synthesis of active proteins containing disulfide bonds 发明人:SWARTZ JAMES ROBERT; KIM DONG-MYUNG 权利人:TRUSTESS OF THE LELAND STANFOR 摘要:Compositions and methods are provided for the enhanced in vitro synthesis of polypeptides containing disulfide bonds. In order to improve the performance of in vitro protein synthesis reactions, pre-treatment and redox buffering of the reaction mix is performed in order to optimize the redox potential. Exogenous enzymes that enhance protein folding and disulfide bond formation may also be added to the reaction.
专利号:US-6451543-B1 优先权日:1998-08-31 标题:Lipid matrix-assisted chemical ligation and synthesis of membrane polypeptides 发明人:KOCHENDOERFER GERD G; HUNTER CHRISTIE L; KENT STEPHEN B H; BOTTI PAOLO 权利人:GRYPHON SCIENCES 摘要:The present invention relates to methods and compositions for lipid matrix-assisted chemical ligation and synthesis of membrane polypeptides that are incorporated in a lipid matrix. The invention is exemplified in production of a prefolded membrane polypeptide embedded within a lipid matrix via stepwise chemoselective chemical ligation of unprotected peptide segments, where at least one peptide segment is embedded in a lipid matrix. Any chemoselective reaction chemistry amenable for ligation of unprotected peptide segments can be employed. Suitable lipid matrices include liposomes, micelles, cell membrane patches and optically isotropic cubic lipidic phase matrices. Prefolded synthetic and semi-synthetic membrane polypeptides synthesized according to the methods and compositions of the invention also permit site-specific incorporation of one or more detectable moieties, such as a chromophore, which can be conveniently introduced during synthesis. The methods and compositions of the invention have multiple uses. For example, they can be used to assay ligand binding to membrane polypeptides and domains comprising a receptor, and thus are extremely useful for structure/function studies, drug screening/selection/design, and diagnostics and the like, including high-throughput applications. The methods and compositions of the invention are particularly suited for FRET analyses of previously inaccessible membrane polypeptides.
专利号:US-7541165-B2 优先权日:2001-10-30 标 题 :Molecular detection systems utilizing reiterative oligonucleotide synthesis 发明人:HANNA MICHELLE M 权利人:RIBOMED BIOTECHNOLOGIES INC 摘要:The present invention provides methods for detecting the presence of a target molecule by generating multiple detectable oligonucleotides through reiterative enzymatic oligonucleotide synthesis events on a defined polynucleotide sequence. The methods generally comprise using a nucleoside, a mononucleotide, an oligonucleotide, or a polynucleotide, or analog thereof, to initiate synthesis of an oligonucleotide product that is substantially complementary to a target site on the defined polynucleotide sequence; optionally using nucleotides or nucleotide analogs as oligonucleotide chain elongators; using a chain terminator to terminate the polymerization reaction; and detecting multiple oligonucleotide products that have been synthesized by the polymerase. In one aspect, the invention provides a method for detecting a target protein, DNA or RNA by generating multiple detectable RNA oligoribonucleotides by abortive transcription.
专利号:US-2004054162-A1 优先权日:2001-10-30 标 题:Molecular detection systems utilizing reiterative oligonucleotide synthesis 发明人:HANNA MICHELLE M 摘要:The present invention provides methods for detecting the presence of a target molecule by the use of nucleotide analogs containing moieties that enable detection. Such analogs may be incorporated into nucleic acids. In one embodiment, nucleotide analogs are used in a process generating multiple detectable oligonucleotides through reiterative enzymatic oligonucleotide synthesis events on a defined polynucleotide sequence. The methods generally comprise using a nucleoside, a mononucleotide, an oligonucleotide, or a polynucleotide, or analog thereof, to initiate synthesis of an oligonucleotide product that is substantially complementary to a target site on the defined polynucleotide sequence; optionally using nucleotides or nucleotide analogs as oligonucleotide chain elongators or chain terminators to terminate the polymerization reaction; and detecting multiple oligonucleotide products that have been synthesized by the polymerase.
专利号:US-9732112-B2 优先权日:2010-05-28 标 题:Synthesis of 2′,3′-dideoxynucleosides for automated DNA synthesis and pyrophosphorolysis activated polymerization 发明人:MA ZHAOCHUN; MULLAH KHAIRUZZAMAN BASHAR; EASON ROBERT G 权利人:LIFE TECHNOLOGIES CORP 摘要:Methods for preparation of 2′,3′-dideoxynucleotides support structures, such as 2′,3′-dideoxyguanosine, 2′,3′-dideoxyadenosine, and 3′-deoxythymidine support structures are disclosed. Various methods of using such structures are also provided, such as their use for automated DNA synthesis and pyrophosphorolysis activated polymerization.
专利号:US-9034576-B2 优先权日:2009-09-24 标题 :Systems and methods for measuring translation of target proteins in cells 发明人:SMILANSKY ZEEV; COOPERMAN BARRY S; GOODMAN YALE E 权利人:SMILANSKY ZEEV; COOPERMAN BARRY S; GOODMAN YALE E; ANIMA CELL METROLOGY INC 摘要:The present invention relates to systems and methods for measuring the rate of translation of a target protein in cells, which are based on the detection of translation of one or more predetermined codon pairs during synthesis of the target protein. The detection is provided by a FRET signal emitted from labeled tRNA molecules which are juxtaposed during synthesis of the protein.
[参考文献]: Adrienne Clements, Et Al. Fluorescence-Based Peptide Labeling And Fractionation Strategies For Analysis Of Cysteine-Containing Peptides. Anal Chem. 2005 Jul 15;77(14):4495-502. [参考文献]: Agnese S Minazzo, Et Al. Loop Dynamics Of The Extracellular Domain Of Human Tissue Factor And Activation Of Factor Viia. Biophys J. 2009 Jan;96(2):681-92. [参考文献]: Christopher R Orton, Et Al. Analysis Of Protein Adduction Kinetics By Quantitative Mass Spectrometry: Competing Adduction Reactions Of Glutathione-S-Transferase P1-1 With Electrophiles. Chem Biol Interact. 2007 Jun 30;168(2):117-27. [参考文献]: Jason R Waggoner, Et Al. Phospholamban Inhibits Ca-Atpase Conformational Changes Involving The E2 Intermediate. Biochemistry. 2007 Feb 20;46(7):1999-2009. [参考文献]: Manunya Nuth, Et Al. Iron-Sulfur Cluster Biosynthesis: Characterization Of Iscu-Iscs Complex Formation And A Structural Model For Sulfide Delivery To The [2Fe-2S] Assembly Site. J Biol Inorg Chem. 2009 Aug;14(6):829-39.
合成参考文献
参考文献:10.1093/oxfordjournals.jbchem.a022433 摘要:Takahashi T, Fukukawa C, Naraoka C, Katoh T, Yazawa M. Conformations of Vertebrate Striated Muscle Myosin Monomers in Equilibrium with Filaments. Journal of Biochemistry. 1999 Jul 01;126(1):34–40. doi: 10.1093/oxfordjournals.jbchem.a022433. 参考文献:10.1371/journal.pone.0020073 摘要:Czupryna J, Tsourkas A. Firefly Luciferase and Rluc8 Exhibit Differential Sensitivity to Oxidative Stress in Apoptotic Cells. PLoS ONE. 2011 May 13;6(5):e20073. doi: 10.1371/journal.pone.0020073. 参考文献:10.1006/abbi.1995.9917 摘要:Liu YC, Sowdal LH, Robinson NC. Separation and quantitation of cytochrome c oxidase subunits by Mono-Q fast protein liquid chromatography and C18 reverse-phase high-performance liquid chromatography. Arch Biochem Biophys. 1995 Dec 01;324(1):135–42. doi: 10.1006/abbi.1995.9917. 参考文献:10.1016/0197-0186(93)90130-w 摘要:Roth GA, Gonzalez MD, Monferran CG, De Santis ML, Cumar FA. Myelin basic protein domains involved in the interaction with actin. Neurochem Int. 1993 Nov;23(5):459–65. doi: 10.1016/0197-0186(93)90130-w. 参考文献:10.1021/bi991343g 摘要:Prochniewicz E, Thomas DD. Differences in structural dynamics of muscle and yeast actin accompany differences in functional interactions with myosin. Biochemistry. 1999 Nov 09;38(45):14860–7. doi: 10.1021/bi991343g.