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

Transgenic rabbit with enhanced green fluorescent protein gene marker, and construction method thereof

A technology of green fluorescent protein and green fluorescence, which is applied to transgenic rabbits carrying enhanced green fluorescent protein and its construction field, can solve the problems of affecting the early development of embryos, reducing the birth rate of transgenic animals, failing to obtain transgenic animals, etc. Strong strength and photostability, shortening promoter sequence, increasing birth rate

Active Publication Date: 2017-05-31
NANJING NORMAL UNIVERSITY
View PDF3 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

GFP transgenic rabbits were produced in France in 2000, but so far there has been no report of producing EGFP transgenic rabbits
Compared with GFP, EGFP has enhanced fluorescence intensity and photostability, and is more suitable for in vivo labeling; but strong EGFP expression will affect the early development of embryos, reduce the birth rate of transgenic animals, and even fail to obtain transgenic animals
Therefore, the construction of enhanced green fluorescent protein transgenic animals remains to be studied

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
  • Transgenic rabbit with enhanced green fluorescent protein gene marker, and construction method thereof
  • Transgenic rabbit with enhanced green fluorescent protein gene marker, and construction method thereof
  • Transgenic rabbit with enhanced green fluorescent protein gene marker, and construction method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] (1) Preparation and optimization of EGFP expression sequence

[0027] The structure of pCAG-GFP plasmid (Addgene, http: / / www.addgene.org / 11150 / ) is as follows figure 1 As shown, it includes a composite promoter sequence composed of CMV enhancer, chicken β promoter protein and CAG promoter, EGFP coding cDNA, and rabbit myosin polyA sequence; the pCAG-GFP plasmid was amplified in Escherichia coli and extracted in large quantities.

[0028] To compare the effect of the promoter sequence on embryonic development, two expression constructs were prepared. Conventional promoter expression components (hereinafter referred to as conventional components): pCAG-GFP plasmid was double digested with SpeI+hindIII, and a 3.1kb fragment was recovered, including a composite promoter sequence composed of CMV enhancer, chicken β promoter protein and CAG promoter, EGFP The coding cDNA, rabbit myosin polyA sequence. Optimized promoter expression component (hereinafter referred to as optim...

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 transgenic rabbit with an enhanced green fluorescent protein gene marker, and a construction method thereof. EGFP (Enhanced Green Fluorescent Protein) expressed by the transgenic rabbit marks all cells of a rabbit individual; green fluorescent is emitted under the excitation of blue light, and a prominent genetic marker sample is used for distinguishing a non-marker sample. The construction method comprises the following steps: introducing a recombinant EGFP coding gene optimized by a promoter into a ribbit fertilized ovum to obtain an EGFP transgenic rabbit; carrying out EGFP gene detection or green fluorescent detection on a baby rabbit, wherein a positive result shows that the baby rabbit is a founder; carrying out hybridization on the founder and a normal rabbit to obtain an EGFP transgenic rabbit strain with genetic stability, and irradiating green fluorescent from rabbit tissues and culture cells subjected to line establishment. A promoter sequence is optimized, the EGFP is applied to manufacture the transgenic rabbit, and the rabbit marked by the EGFP can provide a visually visible biomarker sample from a macroscopic (individual) level to a microcosmic (cell) level.

Description

technical field [0001] The invention relates to a transgenic rabbit carrying an enhanced green fluorescent protein marker and a construction method thereof, in particular to constructing an enhanced green fluorescent protein transgenic rabbit through microinjection, and expressing an enhanced green fluorescent protein labeled rabbit individual All cells emit green fluorescence under excitation conditions of blue light. Background technique [0002] Green fluorescent protein (green fluorescent protein, GFP) is a tracer widely used in life science and medical research today. It is encoded by the GFP gene and emits green fluorescence when excited by light in the blue wavelength range. The optical imaging technology based on GFP enables people to directly observe the rich and colorful life phenomena from microcosmic to macrocosmic levels. [0003] GFP consists of 238 amino acids and has a molecular weight of 26.9kDa. It was originally isolated from Aequorea victoria and emits g...

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): A01K67/027C12N15/89
CPCA01K67/0278C12N15/89A01K2227/107A01K2267/0393
Inventor 杜福良安礼友杨澜
Owner NANJING NORMAL UNIVERSITY
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