Supercharge Your Innovation With Domain-Expert AI Agents!

Establishment method for transgenic zebrafish model with glucose-6-phosphate dehydrogenase deficiency

A technology of phosphate dehydrogenase and establishment method, applied in the field of establishment of transgenic zebrafish model

Inactive Publication Date: 2016-09-07
GUIZHOU MEDICAL UNIV
View PDF2 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there is no ideal animal experimental model that can be well realized g6pd Research on the relationship between gene structure change and function

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
  • Establishment method for transgenic zebrafish model with glucose-6-phosphate dehydrogenase deficiency
  • Establishment method for transgenic zebrafish model with glucose-6-phosphate dehydrogenase deficiency
  • Establishment method for transgenic zebrafish model with glucose-6-phosphate dehydrogenase deficiency

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] The invention provides a method for obtaining transgenic zebrafish by microinjecting plasmids into embryos, and constructs gata1- g6pd M118-144 -egfp-PBSK-Isce I Plasmid, the newly constructed plasmid and Isce I enzyme were injected into zebrafish 1-cell stage embryos together by microinjection, reaching gata1 promoter driver mutation g6pd gene and egfp Effects of gene specificity on erythroid cell expression.

[0029] Wherein, the method for preparing the G6PD transgenic zebrafish model comprises the following steps:

[0030] (1) Prepare mutants by overlapping extension site-directed mutagenesis zg6pd M118-144 Gene:

[0031] A. Determine the 40th-48th amino acid of mutagenic zebrafish G6PD by comparing with human G6PD protein sequence. According to the zebrafish cDNA (NCBI Reference Sequence: XM_694076.5) sequence provided by NCBI, two pairs of primers were designed respectively. The upstream of the first pair of primers added BamHI restriction sites and ...

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 an establishment method for a transgenic zebrafish model with glucose-6-phosphate dehydrogenase (g6pd) deficiency. The establishment method comprises the following steps: (1) performing enzyme digestion and connecting on g6pd<M118-144>-egfp-PCS2+ and gata-1-PBSK-Isce I plasmids to establish gata1-g6pd<M118-144>-egfp-PBSK-Isce I plasmids; (2) injecting the gata1-g6pd<M118-144>-egfp-PBSK-Isce I plasmids into an animal pole embryonic disc of zebrafish in a first cell stage in a micro-injection mode; (3) observing that whether green fluorescence is expressed in embryos in a 24hpf stage to screen transgenic embryos, and hatching and culturing the embryos to adult fish to serve as F0 generation transgenic zebrafish; (4) hatching and culturing the embryos which are produced after the copulation of the F0 generation transgenic zebrafish and wild type zebrafish and express the green fluorescence to adult fish to serve as F1 generation transgenic zebrafish; (5) authenticating the expression and insertion situations of the transferred g6pd<M118-144>-egfp gene by observing the EGFP fluorescence situation and a genome PCR method. The establishment method provides possibility for application of the model to screening medicaments for treating the G6PD deficiency.

Description

technical field [0001] The invention relates to a method for establishing a transgenic zebrafish model, belonging to the technical field of genetic engineering. Background technique [0002] Glucose-6-phosphate dehydrogenase deficiency (glucose-6-phosphate dehydrogenase deficiency, referred to as G6PD deficiency), commonly known as favism, is the most common hereditary hemolytic erythrocyte enzyme deficiency in the world. Red blood cells lose their nuclei and ribosomes during maturation, so they cannot synthesize proteins and cannot carry out the citric acid cycle. Therefore, mature red blood cells mainly rely on the pentose phosphate pathway to provide energy. G6PD is the rate-limiting enzyme of the pentose phosphate metabolic pathway. Glucose 6-phosphate is catalyzed by G6PD to produce nicotinamide adenine dinucleotide phosphate (NADPH) and ribose-5-phosphate (ribose-5-phosphate, R5P). NADPH is extremely important to maintain the reduced state of glutathione (GSH) to pr...

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): C12N15/85C12N15/65A01K67/027A01K67/02
CPCC12N15/8509A01K67/02A01K67/0275A01K2217/05A01K2227/40A01K2267/0306C12N15/65C12N2800/106
Inventor 舒莉萍何志旭周艳华庹媛媛夏海雄崔冬冰
Owner GUIZHOU MEDICAL UNIV
Features
  • R&D
  • Intellectual Property
  • Life Sciences
  • Materials
  • Tech Scout
Why Patsnap Eureka
  • Unparalleled Data Quality
  • Higher Quality Content
  • 60% Fewer Hallucinations
Social media
Patsnap Eureka Blog
Learn More

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

© 2025 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com