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

Recombinant plasmid and cell line for monomolecular positioned super-resolution imaging of exosome and applications of recombinant plasmid and cell line

A technology for super-resolution imaging and recombinant plasmids, which is applied in the field of preparation of recombinant plasmids and cell lines, and can solve the problem of biological samples being inactive

Inactive Publication Date: 2019-01-11
DALIAN UNIV OF TECH
View PDF6 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This kind of labeling technology has certain disadvantages. The indirect immunofluorescence technique needs to carry out the sample fixation process for the labeling of biological samples, resulting in the inactivity of the biological samples themselves.

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
  • Recombinant plasmid and cell line for monomolecular positioned super-resolution imaging of exosome and applications of recombinant plasmid and cell line
  • Recombinant plasmid and cell line for monomolecular positioned super-resolution imaging of exosome and applications of recombinant plasmid and cell line
  • Recombinant plasmid and cell line for monomolecular positioned super-resolution imaging of exosome and applications of recombinant plasmid and cell line

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] 1. Acquisition of exosome marker protein CD63 gene and PAmcherry protein gene

[0026] CD63 protein is an iconic tetraspanning protein on the exosome membrane, and both the C-terminus and the N-terminus of the CD63 protein are in the exosome membrane. The PAmcherry protein gene sequence is fused to the 3' end of the CD63 gene fragment by genetic engineering technology, and the C-terminal of the CD63 protein contains the PAmcherry protein sequence, which realizes fluorescent labeling of the CD63 protein. Using the purchased commercial plasmid containing the CD63 target gene fragment (CD63 gene such as SEQ ID No.5) and the PAmcherry target gene fragment (PAmcherry gene such as SEQ ID No.6) as a template, design upstream and downstream primers to amplify by PCR technology CD63 gene fragment containing restriction endonuclease EcoRV, XhoI site and PAmcherry gene fragment containing restriction endonuclease NotI, XhoI site, such as Figure 4 . in:

[0027] The upstream pr...

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 recombinant plasmid and cell line for monomolecular positioned super-resolution imaging of exosome and applications of the recombinant plasmid and the cell line. The recombinant plasmid is formed by fusing and constructing a fluorescence protein gene with a photoactivation property and a marking CD63 protein gene on an exosome film; Hela cells are transfected through a liposome-mediated plasmid transfection technology; and the Hela cells capable of stably expressing the photoactivation fluorescence protein marked exosome are obtained through drug screening. Monomolecular positioned super-resolution imaging researches of the separated and purified exosome and CD63 protein distribution in cells are carried out, with the help of the photoactivation fluorescence protein. Compared with a traditional microscope, the monomolecular positioned super-resolution imaging provides a better space resolution ratio and the monomolecular positioned super-resolution imaging ofthe exosome with activity is realized; and meanwhile, monomolecular positioned imaging researches of the CD63 protein distribution in the cells are also realized.

Description

technical field [0001] The invention belongs to the technical field of biomarkers, and in particular relates to a preparation method and application of a recombinant plasmid and a cell line for super-resolution imaging of exosome single-molecule positioning. Background technique [0002] Exosomes are small extracellular vesicles formed by cells through the process of "endocytosis-fusion-efflux". Lipid bilayer-structured vesicles with a diameter of 30-150 nm. As an important intercellular information transfer molecule and genetic material transfer carrier, exosomes contain a variety of proteins and RNA substances, which are widely distributed in human blood, urine, saliva and other body fluids as well as tumor cell culture fluid. Exosomes secreted by tumor cells can promote tumor angiogenesis, and can also affect tumor growth and metastasis by regulating the tumor microenvironment. Exosomes play an important role in the development of tumors. Exosome-related fluorescence im...

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): C12N15/63C12N5/10G01N21/64
CPCC07K14/43595C07K14/70596C07K2319/00C12N15/63C12N2510/00G01N21/6428
Inventor 肖义李宁叶智伟陈令成张新富
Owner DALIAN UNIV OF 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