Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Tryptophan optical probe as well as preparation method and application thereof

An optical probe, tryptophan technology, applied in the field of optical probes, can solve the problems of inability to monitor the change of tryptophan concentration in living cells in real time, complicated steps, and prone to human errors.

Pending Publication Date: 2021-09-03
EAST CHINA UNIV OF SCI & TECH
View PDF5 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These detection and analysis methods may require professional analytical equipment, or the steps are cumbersome, prone to human error, and are only suitable for in vitro detection, and cannot monitor the change of tryptophan concentration in living cells in real time

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Tryptophan optical probe as well as preparation method and application thereof
  • Tryptophan optical probe as well as preparation method and application thereof
  • Tryptophan optical probe as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0139] Example 1: Tryptophan-binding protein particles

[0140] The 3lft gene in the Streptococcus pneumoniae gene was amplified by PCR, and the PCR product was recovered after gel electrophoresis and digested with HindIII and XhoI, and the pCDF vector was subjected to corresponding double digestion. After ligation with T4 DNA ligase, the product was used to transform DH5α, and the transformed DH5α was spread on LB plates (streptomycin 100ug / mL), and cultured at 37°C overnight. The growing DH5α transformant was extracted from the plasmid and then identified by PCR. After the positive plasmid is correctly sequenced, the subsequent plasmid construction is carried out.

Embodiment 2

[0141] Example 2: Expression and detection of cpYFP optical probes at different insertion sites

[0142] In this example, based on pCDF-3lft, the following sites were selected to insert cpYFP according to the crystal structure of tryptophan-binding protein, and the corresponding pCDF-3lft-cpYFP plasmids were obtained: 114 / 115, 114 / 116, 114 / 117, 114 / 118, 115 / 116, 115 / 117, 115 / 118, 116 / 117, 116 / 118, 117 / 118, 233 / 234, 233 / 235, 234 / 235, 263 / 264, 263 / 265, 263 / 266 , 263 / 267, 263 / 268, 264 / 265, 264 / 266, 264 / 267, 264 / 268, 265 / 266, 265 / 267, 265 / 268, 266 / 267, 266 / 268 or 267 / 268. Exemplary sequences are shown in Table 1.

[0143] Table 1. Sequences of Optical Probes

[0144] sequence insertion site SEQ ID NO:6 263 / 265 SEQ ID NO:7 263 / 266 SEQ ID NO:8 263 / 267 SEQ ID NO:9 263 / 268

[0145] The cpYFP DNA fragment was generated by PCR, and the cpYFP terminal homologous sequence was introduced through the 5' end of the primer, and the pCDF-tryptophan b...

Embodiment 3

[0148] Example 3: Expression and detection of cpGFP optical probes at different insertion sites

[0149] According to the method in Example 2, cpYFP was replaced with cpGFP to construct a tryptophan green fluorescent protein fluorescent probe. Such as image 3 As shown, the detection results of the broken supernatant show that the optical probes that respond to tryptophan more than 1.5 times are implemented at the 263 / 265, 263 / 266, 263 / 267 and 263 / 268 positions or the corresponding amino acid positions of their family proteins Insert the optical probe.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention relates to a tryptophan optical probe as well as a preparation method and application thereof. In one aspect, the invention relates to an optical probe. The optical probe comprises a tryptophan-sensitive polypeptide or functional variant thereof and an optically active polypeptide or functional variant thereof, wherein the optically active polypeptide or functional variant thereof is located within the sequence of the tryptophan-sensitive polypeptide or functional variant thereof. The invention also relates to a preparation method of the probe and application of the probe in tryptophan detection.

Description

technical field [0001] The invention relates to the technical field of optical probes, in particular to a tryptophan optical probe and its preparation method and application. Background technique [0002] L-tryptophan is one of the essential amino acids indispensable for human growth and development. Among the 20 kinds of amino acids that make up proteins, tryptophan occupies the smallest proportion in proteins and cytoplasm. In addition to regulating protein synthesis, it and its metabolites also participate in multiple physiological functions, and are important for the development of many organs in the body. And functionality is very important. [0003] The metabolism of tryptophan in different tissues is related to various physiological functions. The kynurenine pathway is the main pathway for tryptophan metabolism in humans and mammals, mostly in the liver. In this pathway, tryptophan generates N under the action of tryptophan 2.3-dioxygenase (TDO) (mainly located in ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/62C09K11/06G01N33/68G01N33/58
CPCC07K14/3156C09K11/06G01N33/6812G01N33/582C07K2319/60C09K2211/10C12N15/70C12N15/85C12N15/63
Inventor 杨弋赵玉政张晨霞黄立邹叶君李写
Owner EAST CHINA UNIV OF SCI & TECH
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com