公开(公告)号：US20230296616A1

公开(公告)日：2023-09-21

申请号：US17904498

申请日：2021-02-09

Applicant: East China University of Science and Technology

Inventor： Yi Yang ,  Yuzheng Zhao ,  Rui Li ,  Yejun Zou ,  Li Huang ,  Xie Li ,  Kun Jiang

IPC: G01N33/68 ,  G01N33/533 ,  C07K14/00

CPC classification number: G01N33/6812 ,  G01N33/533 ,  C07K14/001

Abstract: Provided is an arginine fluorescent probe, comprising a polypeptide B that responds to arginine and a fluorescent protein A that expresses arginine; the fluorescent protein A is inserted into the polypeptide B, B is divided into an upper structural part and a lower structural part, B1 and B2, and a probe structure represented by the formula B1-A-B2 is formed; optimized mutants are likewise obtained by truncation and site-directed mutagenesis at different positions, and specific binding of the polypeptide B and arginine leads to a change in the fluorescence signal of the fluorescent protein A; and the polypeptide B is an arginine binding protein or a mutant thereof. The arginine fluorescent probe provided by the present invention has a relatively small protein molecular weight, is easy to express, experiences large dynamic changes in fluorescence, has good specificity, can be expressed, by means of genetic manipulation, in different subcellular organelles of cells, and can be used for the high-throughput, quantitative detection of arginine inside and outside of cells.