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-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:US-2025049961-A1 优先权日:2021-12-23 标题 :Scalable and high-purity cell-free synthesis of closed-ended dna vectors 发明人:MONDS RUSSELL; CIPI JORIS; BLACKSTOCK DANIEL JASON; DURANT JOHN CHESTER 权利人:GENERATION BIO CO 摘要:This disclosure provides methods for scalable and high-purity cell-free synthesis of DNA vectors, particularly closed-ended DNA vectors (e.g., ceDNA vectors) having linear and continuous structure for delivery and expression of a transgene. The cell-free synthesis includes digesting a double-stranded DNA construct with at least one restriction endonuclease that is capable of cleaving the construct at cleavage sites, which are distinct from the recognition sites, to release an insert having unique overhangs that regulate the high specificity of the subsequent ligation reaction. The insert is then ligated with inverted terminal repeat (ITR) oligonucleotides to form the closed-ended DNA vector. Corresponding DNA vectors prepared by these methods and related products as well other base and intermediate vectors and constructs associated with the methods are also provided in this disclosure.
专利号:US-5571677-A 优先权日:1993-07-02 标题:Convergent synthesis of branched and multiply connected macromomolecular structures 发明人:GRYAZNOV SERGEI M 权利人:LYNX THERAPEUTICS INC 摘要:The invention provides methods and compositons for convergent synthesis of branched polymers useful as molecular probes. The invention also includes several novel branched polymeric structures particularly useful for detecting target polynucleotides. Branched polymers of the invention comprise at least two branches: at least one branch is a target binding moiety capable of specifically binding to a target molecule of interest and one or more branches are signal generation moities capable of directly or indirectly generating a detectable signal. In accordance with the method of the invention branched polymers are assembled from components having phosphorothioate groups and/or haloacyl- or haloalkylamino groups. The phosphorothioate and haloacyl- or haloalkylamino groups react rapidly and efficiently when brought into contact to form thiophosphorylacyl- or thiophosphorylalkylamino bridges which.complete the assembly of a branched polymer. The method of the invention permits thorough purification and isolation of the components prior to final assembly. Incomplete branched polymers are readily separated from the desired product by standard means. The method of the invention permits the synthesis of several novel branched polymer configurations.
专利号:WO-2024047178-A1 优先权日:2022-09-02 标题 :A method for simultaneous synthesis of a plurality of oligonucleotides 发明人:ZIV OMER; THORPE CAMERON; TANPURE ARUN; ERLICH YANIV 权利人:ELEVEN THERAPEUTICS LTD 摘要:Disclosed is a method and compounds useful for performing said method for simultaneous synthesis of a plurality of oligonucleotide molecules, and more specifically for orthogonal synthesis of at least two different oligonucleotide molecules at the same time with applications in combinatorial chemistry and DNA encoded libraries (DEL).
参考文献:10.1016/j.brainres.2010.02.025 摘要:Hackman JC, Holohean AM. The effects of polyamine agonists and antagonists on N-methyl-D-aspartate-induced depolarizations of amphibian motoneurons in situ. Brain Res. 2010 Apr 14;1325():10–8. doi: 10.1016/j.brainres.2010.02.025. 参考文献:10.4196/kjpp.2008.12.2.59 摘要:Kim YC, Sim JH, Choi W, Kim CH, You R, Xu W, Lee SJ. Relaxant Effect of Spermidine on Acethylcholine and High K+-induced Gastric Contractions of Guinea-Pig. Korean J Physiol Pharmacol. 2008;12(2):59. doi: 10.4196/kjpp.2008.12.2.59. 参考文献:10.3390/ijms12063723 摘要:Arimura G, Ozawa R, Maffei ME. Recent advances in plant early signaling in response to herbivory. Int J Mol Sci. 2011;12(6):3723–39. 参考文献:10.3390/ijms12063846 摘要:Dai XH, Zhang P, Xiao MF, Zhou RR, Zhang BX, Hu GS, Huang ZB, Fan XG. Protective role of α2HS-glycoprotein in HBV-associated liver failure. Int J Mol Sci. 2011;12(6):3846–56. 参考文献:10.1007/s11010-011-0949-4 摘要:Kreutzer JN, Olsen BB, Lech K, Issinger O, Guerra B. Role of polyamines in determining the cellular response to chemotherapeutic agents: modulation of protein kinase CK2 expression and activity. Molecular and Cellular Biochemistry. 2011 Jul 13;356(1-2):149. doi: 10.1007/s11010-011-0949-4.