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Method for regulating SGLT1 (sodium dependent glucose transporter) and GLUT5 (facilitated glucose transporter 5) expression in chicken intestines through transcription factor GATA3

A sugar transporter and transcription factor technology, applied in biochemical equipment and methods, recombinant DNA technology, and the use of vectors to introduce foreign genetic material, etc., can solve the problems of lack of understanding of the function and regulation mechanism of GATA3 transcription factors, and achieve simple sequencing work Effect

Inactive Publication Date: 2016-04-06
SHANXI AGRI UNIV
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Problems solved by technology

Although GATA3 is expressed in intestinal epithelial cells, the function and regulatory mechanism of GATA3 transcription factors in intestinal epithelial cells are poorly understood, and further studies are needed

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  • Method for regulating SGLT1 (sodium dependent glucose transporter) and GLUT5 (facilitated glucose transporter 5) expression in chicken intestines through transcription factor GATA3
  • Method for regulating SGLT1 (sodium dependent glucose transporter) and GLUT5 (facilitated glucose transporter 5) expression in chicken intestines through transcription factor GATA3
  • Method for regulating SGLT1 (sodium dependent glucose transporter) and GLUT5 (facilitated glucose transporter 5) expression in chicken intestines through transcription factor GATA3

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[0032] A method for the transcription factor GATA3 to regulate the expression of sugar transporters SGLT1 and GLUT5 in the chicken intestine. First, the epithelial cells of the chicken small intestine are isolated, cultured and identified in vitro, and the cultured cells are obtained, and then the 5′ upstream of the SGLT1 and GLUT5 genes are searched using bioinformatics software. The binding site of GATA3 gene, the eukaryotic expression vector pcDNA3.1-GATA3 of transcription factor GATA3 was constructed, and the eukaryotic expression vector pcDNA3.1-GATA3 was transfected into chicken small intestinal epithelial cells by lipofection method, and transfected Afterwards, the chicken small intestinal epithelial cells were collected and cDNA was obtained, and finally the absolute expression levels of the target genes GATA3, SGLT1 and GLUT5 in the cDNA were detected by real-time fluorescent quantitative PCR.

[0033] Specifically include the following steps:

[0034] 1. In vitro iso...

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Abstract

The invention belongs to the technical field of molecular biology and genetic engineering, and aims at providing a method for regulating SGLT1 (sodium dependent glucose transporter) and GLUT5 (facilitated glucose transporter 5) expression in chicken intestines through a transcription factor GATA3. The method comprises the steps of performing separation and cultivation in vitro on chicken intestinal epithelial cells to obtain culture cells, searching GATA3 gene binding sites of SGLT1 gene 5minute and GLUT5 gene 5minute upstream by using bioinformatics software, constructing a eukaryotic expression vector pcDNA 33.1-GATA3, transfecting the eukaryotic expression vector to the chicken intestinal epithelial cells, collecting the cells, obtaining cDNA, and detecting the absolute expression quantity of target genes GATA3, SGLT1 and GLUT5 by real-time fluorescent quantitative PCR. According to the method provided by the invention, information of partial regulation of gene expression is provided for the research on nutrition and breeding of poultry, a theoretical foundation is provided for explaining an interaction mechanism among various genes in a glucose metabolism process, a research basis on a biomolecule level is provided for breeding excellent chicken breeds with a genetic characteristic of high feed conversion ratio, and moreover, a theory reference is provided for breeding practice with the purpose of improving the feed conversion ratio.

Description

technical field [0001] The invention belongs to the technical fields of molecular biology and genetic engineering, and in particular relates to a method for regulating the expression of sugar transporters SGLT1 and GLUT5 in chicken intestinal tract by transcription factor GATA3. Background technique [0002] Monosaccharides are indispensable nutrients for animal growth and development. It is mainly produced by the gluconeogenesis pathway, and some exogenous monosaccharides are also absorbed by the intestine. Dietary soluble carbohydrates are digested in the gut and eventually broken down into simple sugars such as glucose, fructose, and galactose. As the main energy substance in the body, carbohydrates have a vital impact on the metabolism of the body and the homeostasis of the internal environment. Therefore, the study of intestinal carbohydrate absorption mechanism is of great significance to animal nutrition and health, and is a hot research topic at home and abroad. A...

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Application Information

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IPC IPC(8): C12N15/85C12N15/67
CPCC12N15/67C12N15/85C12N2800/107
Inventor 张利环李慧锋朱芷葳张瑜李玲香
Owner SHANXI AGRI UNIV
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