公开(公告)号：US09862942B2

公开(公告)日：2018-01-09

申请号：US14411892

申请日：2013-06-17

Applicant: Japan Science and Technology Agency

Inventor： Tetsuya Yomo ,  Tomoaki Matsuura ,  Haruka Soga ,  Hajime Watanabe ,  Satoshi Fujii

IPC: C12N15/11 ,  C12P21/02 ,  C12N15/10 ,  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.

公开(公告)号：US20150176004A1

公开(公告)日：2015-06-25

申请号：US14411892

申请日：2013-06-17

Applicant: Japan Science and Technology Agency

Inventor： Tetsuya Yomo ,  Tomoaki Matsuura ,  Haruka Soga ,  Hajime Watanabe ,  Satoshi Fujii

IPC: C12N15/11 ,  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中膜蛋白的筛选/选择的效率（例如酶进化法）。 上述目的通过提供一种单层脂质体来实现，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，所述膜蛋白是转运蛋白，所述单层脂质体还包含（e）结合由所述膜蛋白转运的配体的因子。