Fluorescent protein, fusion protein, isolated nucleic acid, vector and application thereof

A fluorescent protein and nucleic acid technology, applied in the field of biomedical optics and molecular imaging, can solve the problems of difficult to achieve simultaneous imaging of multiple fluorescent proteins, difficult to achieve synchronous control, and high cost, to reduce crosstalk, reduce high requirements, and improve The effect of sensitivity

Active Publication Date: 2018-09-07
SHENZHEN INST OF ADVANCED TECH CHINESE ACAD OF SCI
View PDF3 Cites 10 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in multiphoton imaging, adding a secondary Ti:sapphire laser or optical parametric oscillator is expensive and difficult to tune together, so simultaneous imaging of multiple fluorescent proteins is difficult to achieve
[0004] Current micr...

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
  • Fluorescent protein, fusion protein, isolated nucleic acid, vector and application thereof
  • Fluorescent protein, fusion protein, isolated nucleic acid, vector and application thereof
  • Fluorescent protein, fusion protein, isolated nucleic acid, vector and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] The amino acid sequence of the fluorescent protein provided in this example is shown in SEQ ID NO: 2, which includes the following amino acid mutation site: C158D relative to the fluorescent protein mCrimson (whose amino acid sequence is shown in SEQ ID NO: 1).

[0061] The mutation site C158D, the cysteine ​​at position 158 of the mCrimson amino acid sequence is replaced by aspartic acid, which changes the fluorescence spectrum properties of the fluorescent protein of the present invention and has a larger Stokes shift characteristic , its excitation peak and emission peak are 451nm and 613nm respectively, therefore, this fluorescent protein can be used together with the existing fluorescent protein column such as green fluorescent protein, can be used in single photon or multiphoton microscopic multicolor imaging, can effectively Reducing the crosstalk between the fluorescence spectra of different fluorescent proteins helps to improve the sensitivity of imaging, ensure...

Embodiment 2

[0063] The amino acid sequence of the fluorescent protein provided in this example is shown in SEQ ID NO: 3, which includes the following amino acid mutation sites relative to the fluorescent protein mCrimson: C158D, M61F and F174I.

[0064] The fluorescent protein provided in this embodiment not only has the same effect as the fluorescent protein in Example 1, but also through the introduction of M61F and F174I mutation sites, these two mutation sites reflect the brightness characteristics of the fluorescent protein, making the fluorescent protein provided by the present invention Fluorescent proteins also have higher brightness due to their large Stokes shift.

Embodiment 3

[0066] The fluorescent protein provided in this example has an amino acid sequence as shown in SEQ ID NO: 4, which includes the following amino acid mutation sites relative to the fluorescent protein mCrimson: C158D, R10T, Q23Y, G75D, Q111R, E114V, L121V, Q135R and Y218H .

[0067] The fluorescent protein provided in this example not only has the same effect as the fluorescent protein in Example 1, but also through the introduction of multiple mutation sites R10T, Q23Y, G75D, Q111R, E114V, L121V, Q135R and Y218H, these sites reflect the The maturation rate characteristic of the fluorescent protein makes the fluorescent protein provided by the present invention have a higher maturation rate in terms of the characteristic of a large Stokes shift.

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 discloses a fluorescent protein, a fusion protein, an isolated nucleic acid, a vector and application thereof, belonging to the technical field of biomedical optics and molecular imaging. The fluorescent protein disclosed in the invention further comprises the amino acid mutation site C158D compared with the conventional fluorescent protein mCrimson. The mutation site enables the fluorescence spectral properties of the fluorescent protein of the invention to be changed, and the fluorescent protein has substantial Stokes shift characteristics, is applicable to single-photon or multi-photon microscopic multicolor imaging, can effectively reduce crosstalk between the fluorescence spectra of different fluorescent proteins, enhances the sensitivity of imaging, ensures simultaneousfluorescent imaging of the fluorescent proteins with different spectra and reduces high requirements on optical devices.

Description

technical field [0001] The invention relates to the technical field of biomedical optics and molecular imaging, in particular to a fluorescent protein, fusion protein, isolated nucleic acid, carrier and application. Background technique [0002] When using fluorescent proteins for in vivo imaging, it is often difficult to achieve multi-channel simultaneous imaging. If this limitation can be broken through, the detection of multiple biological rapid response events can be realized, which will greatly promote the development of neuroscience. [0003] In single-photon imaging, multiple fluorescent proteins can be excited by light with different excitation wavelengths to achieve multi-channel simultaneous imaging. However, in multiphoton imaging, the cost of adding a secondary Ti:sapphire laser or optical parametric oscillator is high, and it is difficult to tune them together, so simultaneous imaging of multiple fluorescent proteins is difficult to achieve. [0004] Current mi...

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
IPC IPC(8): C07K14/435C07K19/00C12N15/12G01N21/64
CPCG01N21/6428C07K14/43595C07K2319/00
Inventor 储军郭育奇张书张楚秋刘丰
Owner SHENZHEN INST OF ADVANCED TECH CHINESE ACAD OF SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap