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    Fluorescent Probe for Branched Chain Amino Acids and Use Thereof
    2.
    发明公开
    Fluorescent Probe for Branched Chain Amino Acids and Use Thereof 审中-公开

    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.

    Nicotinamide Adenine Dinucleotide Gene Encoding Fluorescent Probe, Preparation Method Therefor and Application Thereof
    6.
    发明申请
    Nicotinamide Adenine Dinucleotide Gene Encoding Fluorescent Probe, Preparation Method Therefor and Application Thereof 有权
    烟酰胺腺嘌呤二核苷酸基因编码荧光探针及其制备方法及应用

    公开(公告)号：US20140329718A1

    公开(公告)日：2014-11-06

    申请号：US14347575

    申请日：2012-09-26

    Applicant: East China University of Science and Technology

    Inventor： Yi Yang ,  Jing Jin ,  Yuzheng Zhao ,  Qingxun Hu

    IPC: G01N33/573 ,  G01N33/58 ,  G01N33/50

    CPC classification number: G01N33/5735 ,  C07K14/195 ,  C07K14/32 ,  C07K2319/00 ,  C07K2319/60 ,  G01N33/5011 ,  G01N33/582 ,  G01N2500/02 ,  G01N2800/7028

    Abstract: The invention relates to a genetically encoded fluorescent sensor for nicotinamide adenine dinucleotide, as well as methods of preparation and uses thereof. In one aspect, this invention relates to a sensor for detecting nicotinamide adenine dinucleotide, particularly, a recombinant fluorescent fusion protein sensor for detecting nicotinamide adenine dinucleotide. In one specific aspect, this invention relates to a recombinant fluorescent fusion protein sensor for detecting reduced nicotinamide adenine dinucleotide (NADH); in another specific aspect, this invention relates to a recombinant fluorescent fusion protein sensor for detecting oxidized nicotinamide adenine dinucleotide (NAD+); in yet another aspect, the invention relates to a recombinant fluorescent fusion protein sensor for detecting the ratio of reduced to oxidized nicotinamide adenine dinucleotide. This invention also relates to the method of preparing the sensors, and uses of the sensors in detecting NADH, NAD+, NADH/NAD+ ratio, screening drugs and measuring NADH metabolism.

    Abstract translation: 本发明涉及用于烟酰胺腺嘌呤二核苷酸的遗传编码荧光传感器，以及其制备方法和用途。 一方面，本发明涉及用于检测烟酰胺腺嘌呤二核苷酸的传感器，特别是用于检测烟酰胺腺嘌呤二核苷酸的重组荧光融合蛋白传感器。 在一个具体方面，本发明涉及用于检测还原型烟酰胺腺嘌呤二核苷酸（NADH）的重组荧光融合蛋白传感器。 在另一具体方面，本发明涉及用于检测氧化烟酰胺腺嘌呤二核苷酸（NAD +）的重组荧光融合蛋白传感器。 在另一方面，本发明涉及用于检测还原氧化烟酰胺腺嘌呤二核苷酸比例的重组荧光融合蛋白传感器。 本发明还涉及传感器的制备方法，以及传感器检测NADH，NAD +，NADH / NAD +比例，筛选药物和测定NADH代谢的用途。

    ARGININE FLUORESCENT PROBE, PREPARATION METHOD THEREFOR AND APPLICATION THEREOF
    8.
    发明公开
    ARGININE FLUORESCENT PROBE, PREPARATION METHOD THEREFOR AND APPLICATION THEREOF 审中-公开

    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.

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