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Application of transgenic zebrafish model to screening of drugs for treating G6PD deficiency

A zebrafish, transgenic technology, applied in the field of genetic engineering

Inactive Publication Date: 2016-09-21
GUIZHOU MEDICAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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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

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  • Application of transgenic zebrafish model to screening of drugs for treating G6PD deficiency
  • Application of transgenic zebrafish model to screening of drugs for treating G6PD deficiency
  • Application of transgenic zebrafish model to screening of drugs for treating G6PD deficiency

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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 ...

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Abstract

The invention discloses application of a transgenic zebrafish model to screening of drugs for treating G6PD deficiency. According to the transgenic zebrafish model, an erythroid cell specific promoter gatal drives mutant g6pd genes deficient at sites 118-144, and meanwhile the transgenic zebrafish model expresses EGFP and is traceable. The zebrafish model is about glucose-6-phosphate dehydrogenase deficiency (G6PDD), is established through a dominant negative effect and is used for traceable observation of changes of erythrocytes in the drug treatment process to achieve application of the model in the process of screening the drugs for treating G6PD deficiency. A brand new convenient mode for screening the drugs for treating G6PD deficiency is obtained, and the drug screening efficiency is improved.

Description

technical field [0001] The invention relates to the application of a transgenic zebrafish model in screening drugs for treating G6PD deficiency, and belongs 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 t...

Claims

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Application Information

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IPC IPC(8): C12N15/85G01N21/84A01K61/00
CPCA01K61/00A01K2227/40A01K2267/0393C12N15/8509G01N21/84G01N2500/10Y02A40/81
Inventor 何志旭舒莉萍周艳华夏海雄宋锦吴西军杨燕
Owner GUIZHOU MEDICAL UNIV
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