Title:
標的タンパク質に作用するポリペプチドのスクリーニング方法
Document Type and Number:
Japanese Patent JP7412043
Kind Code:
B2
Abstract:
The present invention relates to a method for screening for a polypeptide that acts on a target protein. The present invention relates more specifically, to a method for screening for a polypeptide that acts on a target protein, comprising the steps of:(1) providing a polynucleotide library constituted of a plurality of expression vectors that can be expressed in a gram-negative bacterium, wherein the plurality of expression vectors each comprise a first polynucleotide encoding a polypeptide different from one another, a secretory signal sequence positioned upstream of the first polynucleotide, and a second polynucleotide encoding a target protein;(2) transforming a gram-negative bacterium with the expression vector to express the polypeptide in a periplasmic space and the target protein on an inner membrane surface;(3) contacting the polypeptide with the target protein in the periplasmic space;(4) allowing the gram-negative bacterium to form a spheroplast to measure activity of the polypeptide on the target protein by an electrophysiological technique; and(5) identifying a polypeptide that acts on the target protein based on the measured activity.
Inventors:
Tadashi Kimura
Application Number:
JP2022541588A
Publication Date:
January 12, 2024
Filing Date:
August 04, 2021
Export Citation:
Assignee:
National Institute of Advanced Industrial Science and Technology
International Classes:
C12Q1/02; C07K14/435; C12N1/21; C12N15/70; C12P21/00; C40B40/06; C40B40/08; G01N33/15
Domestic Patent References:
| JP2014207905A | ||||
| JP200811A |
Other References:
KIKUCHI, Kyoko et al.,The application of the Escherichia coli giant spheroplast for drug screening with automated planar p,Biotechnology Reports,2015年,7,17-23
FERNANDEZ-RECIO, Juan et al.,Optimal Docking Area: A New Method for Predicting Protein-Protein Interaction Sites,PROTEINS,2005年,58,134-143
FERNANDEZ-RECIO, Juan et al.,Optimal Docking Area: A New Method for Predicting Protein-Protein Interaction Sites,PROTEINS,2005年,58,134-143
Attorney, Agent or Firm:
Seiji Ohno
Yuko Matsutoya
Shinsuke Umeda
Hiroshi Morita
Yuko Matsutoya
Shinsuke Umeda
Hiroshi Morita