鸡转录因子GATA2影响单糖转运蛋白基因GLUT2、GLUT5和SGLT1表达的研究

发布时间：2018-07-01 09:50

本文选题：GATA2 + 糖类转运结合蛋白　；参考：《山西农业大学》2015年硕士论文

【摘要】：GLUT2、GLUT5和SGLT1这三种营养物质转运蛋白基因的表达直接影响着肠道营养吸收功能。为了从分子水平上揭示转录因子GATA2对以上三种功能基因调控作用。本试验研究了增加GATA2转录因子表达量对三种功能基因表达量的影响情况。首先,利用生物信息学软件ModuleMaster1.4、http://tfbind.hgc.jp等软件预测转录因子GATA2与三种单糖转运蛋白基因GZUT2、GLUT5和SGLT1上游调控区内的调控位点。通过构建重组质粒pcDNA3.1-GATA2。其次,将构建的重组质粒通过脂质体介导法转染入鸡小肠上皮细胞中。最后,通过绝对荧光定量PCR检测转染后不同的三组细胞中GATA2以及单糖转运蛋白基因(GUT2、SGLT1和GLUT5的mRNA表达量。研究发现,试验组的GATA2基因的mRNA表达量与阴性对照组和空白对照组相比高6倍,经数据统计分析试验组与两个对照组分别呈极显著性差异(P0.01)。表明重组质粒pcDNA3.1-GATA2转染成功。试验组的GLUT2基因的mRNA表达量与阴性对照组和空白对照组相比降低。经数据统计分析试验组与两个对照组分别呈极显著性差异(P0.01)。表明GATA2转录因子表达量增加,会抑制单糖转运蛋白基因GLUT2的表达。同样的条件下,三组中SGLT1和GLUT5的mRNA表达量却无明显变化。经数据统计分析试验组与两个对照组分别呈不显著性差异(P0.05)。表明仅增加GATA2转录因子表达量不能影响单糖转运蛋白基因SGLT1、GLUT5表达量变化。本研究结果为进一步研究GATA2转录因子在鸡肠道中的生物学功能和选择适宜的转录结合位点提供理论基础。
[Abstract]:GLUT2, GLUT5 and SGLT1 are three kinds of nutrient transporter genes, which directly affect the nutrient absorption function of intestinal tract. In order to reveal the regulatory effects of GATA2 on the above three functional genes at the molecular level. The effect of increasing GATA2 transcription factor expression on the expression of three functional genes was studied. Firstly, the bioinformatics software, Module Master 1.4 / http / tfbind.hgc.jp, was used to predict the regulatory sites of transcription factor GATA2 and three monosaccharide transporter genes GZUT2GLUT5 and SGLT1. The recombinant plasmid pcDNA3.1-GATA2 was constructed. Secondly, the constructed recombinant plasmid was transfected into chicken intestinal epithelial cells by liposome mediated transfection. Finally, the mRNA expressions of GATA2 and GUT5 (GUT2, SGLT1 and GLUT5) were detected by absolute fluorescence quantitative PCR. It was found that the mRNA expression of GATA2 gene in the experimental group was 6 times higher than that in the negative control group and the blank control group, and there was a significant difference between the experimental group and the two control groups by statistical analysis (P0.01). The recombinant plasmid pcDNA3.1-GATA2 was transfected successfully. GLUT2 mRNA expression in experimental group was lower than that in negative control group and blank control group. There was a significant difference between the experimental group and the two control groups by statistical analysis (P0.01). The results showed that GATA2 transcription factor increased the expression of GLUT2 and inhibited the expression of monosaccharide transporter gene GLUT2. Under the same conditions, the expression of SGLT1 and GLUT5 mRNA did not change significantly in the three groups. There was no significant difference between the experimental group and the two control groups by statistical analysis (P0.05). The results showed that only increasing the expression of GATA2 transcription factor could not affect the expression of SGLT1 and GLUT5 in monosaccharide transporter gene. The results provide a theoretical basis for the further study of the biological function of GATA2 transcription factors in chicken intestinal tract and the selection of appropriate transcription binding sites.
【学位授予单位】：山西农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S831

【参考文献】