公开(公告)号：EP2118129A2

公开(公告)日：2009-11-18

申请号：EP08731178.3

申请日：2008-02-29

申请人： Life Technologies Corporation

发明人： KATZEN, Federico ,  FLETCHER, Julia ,  KUDLICKI, Wieslaw ,  BEECHEM, Joseph ,  WANG, Lilin

IPC分类号： C07K14/47 ,  C07K1/00 ,  C12Q1/68 ,  C12P21/06

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： 提供了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的系统和方法。 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支架蛋白质例如载脂蛋白或含有（AAHC）蛋白质的两亲性α螺旋，其中产生的可溶性蛋白质产率高于 缺少支架蛋白。 可以在与脂质相关的体外蛋白质合成系统中提供支架蛋白质，或者不提供与脂质相关的支架蛋白质。 支架蛋白本身可以作为蛋白质提供，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。