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    Nicotinamide Adenine Dinucleotide Gene Encoding Fluorescent Probe, Preparation Method Therefor and Application Thereof
    3.
    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
    5.
    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.

    Light-switchable gene expression system and the methods for controlling gene expression in prokaryotic bacterium
    6.
    发明授权
    Light-switchable gene expression system and the methods for controlling gene expression in prokaryotic bacterium 有权

    公开(公告)号：US10184126B2

    公开(公告)日：2019-01-22

    申请号：US14419106

    申请日：2013-08-01

    Applicant: EAST CHINA UNIVERSITY OF SCIENCE AND TECHNOLOGY

    Inventor： Yi Yang ,  Xianjun Chen ,  Zhengcai Ma ,  Renmei Liu

    IPC: C12N15/62 ,  C12N15/70 ,  C12N15/74 ,  C12N15/63 ,  C12N15/75 ,  C12P21/02 ,  C12N9/02 ,  C12N13/00

    Abstract: Provided is an optically controlled gene expression system of prokaryotic bacterium, comprising: a) a photosensitive recombinant transcription factor encoding gene, the photosensitive recombinant transcription factor is one fusion protein comprising a first polypeptide as the DNA bonding domain and a second polypeptide as the photosensitive domain; b) a target transcription unit comprising promoter or promoter-reaction element or reaction element-promoter containing at least one reaction element recognized/bound by the first polypeptide and the nucleic acid sequence to be transcribed. Also provided is a prokaryotic expression vector comprising said optically controlled gene expression system, and a method for regulating gene expression in a prokaryotic host cell by using the optically controlled gene expression system. Also provided is a reagent kit containing different components of the optically controlled gene expression system. The optically controlled gene expression system of prokaryotic bacterium has a quick, effective and powerful induction, is safer than other inducers, is of little or no toxicity, and can control gene expression both spatially and temporally, and can regulate many life processes of prokaryotic bacterium.

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