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-12188072-B2 优先权日:2018-03-19 标题 :Compositions and methods for rapid in vitro synthesis of bioconjugate vaccines in vitro via production and N-glycosylation of protein carriers in detoxified prokaryotic cell lysates 发明人:JEWETT MICHAEL CHRISTOPHER; STARK JESSICA CAROL; DELISA MATTHEW P; JAROENTOMEECHAI THAPAKORN 权利人:UNIV NORTHWESTERN; UNIV CORNELL 摘要:Disclosed are methods, systems, components, and compositions for cell-free synthesis of glycosylated carrier proteins. The glycosylated carrier proteins may be utilized in vaccines, including anti-bacterial vaccines. The glycosylated carrier proteins may include a bacterial polysaccharide conjugated to a carrier, which may be utilized to generate an immune response in an immunized host against the polysaccharide conjugated to the carrier. The glycosylated carrier proteins may be synthesized in cell-free glycoprotein synthesis (CFGpS) systems using prokaryote cell lysates that are enriched in components for glycoprotein synthesis such as oligosaccharyltransferases (OSTs) and lipid-linked oligosaccharides (LLOs) including OSTs and LLOs associated with synthesis of bacterial O antigens.
专利号:US-6168931-B1 优先权日:1999-03-17 标题 :Enhanced in vitro synthesis of biological macromolecules using a novel ATP regeneration system 发明人:SWARTZ JAMES R; KIM DONG-MYUNG 权利人:UNIV LELAND STANFORD JUNIOR 摘要:Compositions and methods are provided for the enhanced in vitro synthesis of biological molecules where ATP is required for synthesis. Of particular interest is the synthesis of polymers, e.g. nucleic acids, polypeptides, and complex carbohydrates. A homeostatic system is used for production of ATP, where the required high energy phosphate bonds are generated in situ, e.g. through coupling with an oxidation reaction. The homeostatic energy source will typically lack high energy phosphate bonds itself, and will therefore utilize free phosphate in the reaction mix during generation of ATP. Since inorganic phosphate can be an inhibitory by-product of synthesis, the period of time when synthesis is maintained in vitro can be extended. The homeostatic energy source is provided in combination with an enzyme that catalyzes the creation of high energy phosphate bonds and with an enzyme that can use that high energy phosphate bond to regenerate ATP.
专利号:US-6783957-B1 优先权日:1999-03-25 标题:Method for synthesis of polypeptides in cell-free systems 发明人:BIRYUKOV SERGEY VLADIMIROVICH; SIMONENKO PETER NIKOLAEVICH; SHIROKOV VLADIMIR ANATOLIEVICH; SPIRIN ALEXANDER SERGEYEVICH 权利人:ROCHE DIAGNOSTICS GMBH; INST OF PROTEIN RES 摘要:The method of polypeptide synthesis in eukaryotic or porkaryotic cell-free systems based on a modified verion of synthesis in the continuous flow or continuous exchange modes when, in addition to input into the reaction mixture of components maintaining the synthesis and removal from the reaction mixture of low molecular weight components inhibiting the synthesis, the concentration of at least one of the components selected from the group consisting of Mg 2+ , K + , NTP, polyamines or their combinations determining the productivity of the synthesis is continuously changed within the given range of concentrations, while the concentrations of the other components are maintained constant.
专利号:EP-1309698-B1 优先权日:2000-07-21 标 题:A cytochrome p450 enzyme associated with the synthesis of delta-12 -epoxy groups in fatty acids of plants 发明人:CAHOON EDGAR B 权利人:DU PONT 摘要:An isolated nucleic acid fragment encoding a cytochrome P450 enzyme associated with the synthesis of plant DELTA12-epoxy fatty acids is disclosed. The invention also relates to making a chimeric construct encoding all or a portion of the cytochrome P450 enzymes associated with the synthesis of plant DELTA12-epoxy fatty acids, in sense or antisense orientation, wherein expression of the chimeric construct results in production of altered levels of the cytochrome P450 enzymes associated with the synthesis of plant DELTA12-epoxy fatty acids in a transformed host cell.
专利号:US-12421628-B2 优先权日:2018-07-23 标题:Massively parallel enzymatic synthesis of nucleic acid strands 发明人:HORGAN ADRIAN; GODRON XAVIER; YBERT THOMAS; NICOL ROBERT 权利人:DNA SCRIPT 摘要:The invention is directed to methods for massively parallel template-free enzymatic synthesis of a plurality of different polynucleotides of predetermined sequences. In one aspect, methods of the invention employ large scale arrays of reaction sites each associated with at least one working electrode for controlling deprotection and deblocking steps at predetermined user selected sites. In another aspect, the invention provides template-free enzymatic synthesis with proofreading, wherein completed polynucleotides at predetermined reaction sites are sequenced using a sequencing by synthesis technique, particularly employing electrochemically labile blocking groups.
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合成参考文献
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