公开(公告)号：EP2876159B1

公开(公告)日：2019-08-07

申请号：EP13810803.0

申请日：2013-06-17

Applicant: Japan Science and Technology Agency

Inventor： YOMO, Tetsuya ,  MATSUURA, Tomoaki ,  SOGA, Haruka ,  WATANABE, Hajime ,  FUJII, Satoshi

IPC: C12N15/09 ,  C12P21/02 ,  C12Q1/68

公开(公告)号：EP2876159A1

公开(公告)日：2015-05-27

申请号：EP13810803.0

申请日：2013-06-17

Applicant: Japan Science and Technology Agency

Inventor： YOMO, Tetsuya ,  MATSUURA, Tomoaki ,  SOGA, Haruka ,  WATANABE, Hajime ,  FUJII, Satoshi

IPC: C12N15/09 ,  C12P21/02 ,  C12Q1/68

CPC classification number: C12N15/11 ,  C12N15/102 ,  C12N15/1034 ,  C12P21/02 ,  C12Q1/686

Abstract: The objective of the present invention is to improve the efficiency of screening/selection of a membrane protein in molecular evolutionary engineering (for example, an enzyme evolutionary method).
The above-described objective is achieved by providing a unilamellar liposome comprising:
(a) a DNA comprising a promoter sequence, a translational initiation sequence, and a sequence encoding a membrane protein;
(b) an RNA polymerase;
(c) a ribonucleotide; and
(d) a cell-free protein synthesis system. In one aspect of the present invention, the membrane protein is a transporter, and the unilamellar liposome further comprises
(e) a factor that binds to a ligand transported by the membrane protein.

Abstract translation: 本发明的目的是提高分子进化工程（例如酶进化方法）中筛选/选择膜蛋白的效率。 上述目的通过提供单层脂质体来实现，所述单层脂质体包含：（a）包含启动子序列，翻译起始序列和编码膜蛋白的序列的DNA; （b）RNA聚合酶; （c）核糖核苷酸; 和（d）无细胞蛋白质合成系统。 在本发明的一个方面中，膜蛋白是转运蛋白，并且单层脂质体还包含（e）结合由膜蛋白转运的配体的因子。