公开(公告)号：US20240027344A1

公开(公告)日：2024-01-25

申请号：US16981088

申请日：2019-03-15

Applicant: East China University of Science and Technology

Inventor： Yi Yang ,  Yuzheng Zhao ,  Xie Li ,  Yejun Zou ,  Changcheng Zhang

IPC: G01N21/64

CPC classification number: G01N21/6428

Abstract: The disclosure provides a fluorescent sensor, comprising a) a responsive polypeptide, and b) an optically active polypeptide, wherein the optically active polypeptide is inserted into the responsive polypeptide. The present disclosure also relates to a nucleic acid sequence encoding the fluorescent sensor according to any embodiment, or a complementary sequence thereof. The disclosure also provides an expression vector comprising the nucleic acid sequence or a complementary sequence thereof of the disclosure operably linked to an expression control sequence. The disclosure also provides a cell containing the expression vector of the disclosure. The present disclosure also provides a method for preparing the fluorescent sensor of this disclosure, comprising the following steps: providing a host cell comprising an expression vector that expresses the fluorescent sensor of the disclosure, culturing the host cell under conditions suitable for the expression of the host cell, and separating the fluorescent sensor. The disclosure also provides uses of the fluorescent sensor described in the disclosure or the fluorescent sensor prepared according to the method of the disclosure in detecting BCAAs. In one embodiment, the branched chain amino acids are selected from leucine, isoleucine and valine. The detection can be conducted in vitro, in vivo, in situ, or at subcellular level. The disclosure also provides a kit, comprising the fluorescent sensor described in the disclosure or the fluorescent sensor prepared according to the method of the disclosure.

公开(公告)号：US20230296617A1

公开(公告)日：2023-09-21

申请号：US17904503

申请日：2021-02-09

Applicant: East China University of Science and Technology

Inventor： Yi Yang ,  Yuzheng Zhao ,  Chenxia Zhang ,  Li Huang ,  Yejun Zou ,  Xie Li ,  Shuning Liu

IPC: G01N33/68 ,  G01N33/58 ,  C12N15/63

CPC classification number: G01N33/6812 ,  G01N33/582 ,  C12N15/63

Abstract: Disclosed is a tryptophan optical probe, a preparation method therefor and the use thereof. Disclosed is an optical probe, comprising a tryptophan-sensitive polypeptide or a functional variant thereof and an optically active polypeptide or a functional variant thereof, wherein the optically active polypeptide or the functional variant thereof is located in the sequence of the tryptophan-sensitive polypeptide or the functional variant thereof. Furthermore, disclosed are a method for preparing the above-mentioned probe and the use thereof in the detection of tryptophan.

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.