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1. 发明申请

US20110104781A1 ISOLATED PHOSPHOLIPID-PROTEIN PARTICLES 审中-公开
标题翻译：分离的磷脂 - 蛋白质颗粒
公开(公告)号：US20110104781A1
公开(公告)日：2011-05-05
申请号：US12890386
申请日：2010-09-24
申请人： FEDERICO KATZEN , JULIA FLETCHER , WIESLAW KUDLICKI , JOSEPH BEECHEM , LILIN WANG
发明人： FEDERICO KATZEN , JULIA FLETCHER , WIESLAW KUDLICKI , JOSEPH BEECHEM , LILIN WANG
IPC分类号： C12N9/00 , C07K14/00 , C12N5/071 , C12N1/16
CPC分类号： C12P21/02 , C07K1/02
摘要： Systems and methods are provided for producing a protein of interest that is typically not amenable to expression in soluble form in in vitro expression systems. In some aspects, the invention provides methods of synthesizing proteins using in vitro protein synthesis systems that include a scaffold protein such as apolipoprotein or an amphipathic alpha helix containing (“AAHC”) protein, in which higher yields of soluble protein are produced than in the absence of the scaffold protein. The scaffold proteins may be provided in an in vitro protein synthesis system associated with lipid or not associated with lipid. The scaffold protein may be provided as a protein per se or may be encoded by a nucleic acid template and co-expressed with the protein of interest. The invention also provides compositions and kits for synthesis of proteins in soluble form, in which the compositions and kits include cell extracts for protein expression and isolation.
摘要翻译：提供的系统和方法用于产生通常不适于在体外表达系统中以可溶形式表达的目的蛋白质。在一些方面，本发明提供了使用体外蛋白质合成系统合成蛋白质的方法，所述体外蛋白质合成系统包括诸如载脂蛋白或含有（“AAHC”）蛋白质的两亲性α螺旋的支架蛋白，其中产生比在缺少支架蛋白。支架蛋白质可以提供在与脂质相关的体外蛋白质合成系统中或与脂质不相关。支架蛋白质可以作为蛋白质本身提供，或者可以由核酸模板编码并与感兴趣的蛋白质共表达。本发明还提供用于合成可溶形式的蛋白质的组合物和试剂盒，其中组合物和试剂盒包括用于蛋白质表达和分离的细胞提取物。

2. 发明申请

US20110195450A1 In Vitro Protein Synthesis Systems for Membrane Proteins that Include Adolipoproteins and Phospholipid Adolipoprotein Particles 审中-公开
标题翻译：包含载脂蛋白和磷脂脂蛋白颗粒的膜蛋白的体外蛋白质合成系统
公开(公告)号：US20110195450A1
公开(公告)日：2011-08-11
申请号：US12848017
申请日：2010-07-30
申请人： WIESLAW KUDLICKI , JULIA FLETCHER , FEDERICO KATZEN
发明人： WIESLAW KUDLICKI , JULIA FLETCHER , FEDERICO KATZEN
IPC分类号： C12P21/00 , C12N9/00
CPC分类号： C12P21/00
摘要： In vitro protein synthesis systems and methods are provided that produce membrane proteins in soluble form. In some aspects, the invention provides methods of synthesizing proteins using in vitro protein synthesis systems that include an apolipoprotein, in which higher yields of soluble protein are produced than in the absence of the apolipoprotein. Apolipoproteins useful in the present invention include naturally occurring apolipoproteins, as well as sequence variants of wild-type apolipoproteins, and engineered apolipoproteins. The apolipoproteins can be provided in an in vitro protein synthesis system associated with lipid or not associated with lipid. The invention also provides compositions and kits for synthesis of proteins in soluble form, in which the compositions and kits include cell extracts for protein translation and at least one apolipoprotein biomolecule.
摘要翻译：提供体外蛋白质合成系统和方法，其产生可溶形式的膜蛋白质。在一些方面，本发明提供使用体外蛋白质合成系统合成蛋白质的方法，所述体外蛋白质合成系统包括与不存在载脂蛋白时产生更高产率的可溶性蛋白质的载脂蛋白。可用于本发明的载脂蛋白包括天然存在的载脂蛋白，以及野生型载脂蛋白和工程化载脂蛋白的序列变体。载脂蛋白可以在与脂质相关的体外蛋白质合成系统中提供，或与脂质不相关。本发明还提供用于合成可溶形式的蛋白质的组合物和试剂盒，其中组合物和试剂盒包括用于蛋白质翻译的细胞提取物和至少一种载脂蛋白生物分子。

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