本文選題：GATA2 + 糖類轉(zhuǎn)運結(jié)合蛋白�。� 參考：《山西農(nóng)業(yè)大學(xué)》2015年碩士論文

【摘要】：GLUT2、GLUT5和SGLT1這三種營養(yǎng)物質(zhì)轉(zhuǎn)運蛋白基因的表達直接影響著腸道營養(yǎng)吸收功能。為了從分子水平上揭示轉(zhuǎn)錄因子GATA2對以上三種功能基因調(diào)控作用。本試驗研究了增加GATA2轉(zhuǎn)錄因子表達量對三種功能基因表達量的影響情況。首先,利用生物信息學(xué)軟件ModuleMaster1.4、http://tfbind.hgc.jp等軟件預(yù)測轉(zhuǎn)錄因子GATA2與三種單糖轉(zhuǎn)運蛋白基因GZUT2、GLUT5和SGLT1上游調(diào)控區(qū)內(nèi)的調(diào)控位點。通過構(gòu)建重組質(zhì)粒pcDNA3.1-GATA2。其次,將構(gòu)建的重組質(zhì)粒通過脂質(zhì)體介導(dǎo)法轉(zhuǎn)染入雞小腸上皮細胞中。最后,通過絕對熒光定量PCR檢測轉(zhuǎn)染后不同的三組細胞中GATA2以及單糖轉(zhuǎn)運蛋白基因(GUT2、SGLT1和GLUT5的mRNA表達量。研究發(fā)現(xiàn),試驗組的GATA2基因的mRNA表達量與陰性對照組和空白對照組相比高6倍,經(jīng)數(shù)據(jù)統(tǒng)計分析試驗組與兩個對照組分別呈極顯著性差異(P0.01)。表明重組質(zhì)粒pcDNA3.1-GATA2轉(zhuǎn)染成功。試驗組的GLUT2基因的mRNA表達量與陰性對照組和空白對照組相比降低。經(jīng)數(shù)據(jù)統(tǒng)計分析試驗組與兩個對照組分別呈極顯著性差異(P0.01)。表明GATA2轉(zhuǎn)錄因子表達量增加,會抑制單糖轉(zhuǎn)運蛋白基因GLUT2的表達。同樣的條件下,三組中SGLT1和GLUT5的mRNA表達量卻無明顯變化。經(jīng)數(shù)據(jù)統(tǒng)計分析試驗組與兩個對照組分別呈不顯著性差異(P0.05)。表明僅增加GATA2轉(zhuǎn)錄因子表達量不能影響單糖轉(zhuǎn)運蛋白基因SGLT1、GLUT5表達量變化。本研究結(jié)果為進一步研究GATA2轉(zhuǎn)錄因子在雞腸道中的生物學(xué)功能和選擇適宜的轉(zhuǎn)錄結(jié)合位點提供理論基礎(chǔ)。
[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.
【學(xué)位授予單位】：山西農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S831

【參考文獻】

相關(guān)期刊論文 前10條

1 劉芳寧;張彥明;;哺乳動物腸上皮細胞的原代培養(yǎng)[J];動物醫(yī)學(xué)進展;2007年04期

2 趙小剛,安娟;葡萄糖轉(zhuǎn)運蛋白的研究進展[J];飼料工業(yè);2005年14期

3 祁建生,廖承濟;葡萄糖轉(zhuǎn)運體家族分子生物學(xué)研究進展[J];福建中醫(yī)學(xué)院學(xué)報;2000年02期

4 翁梅倩;腸發(fā)育與糖消化、吸收的分子學(xué)研究進展[J];國外醫(yī)學(xué)(兒科學(xué)分冊);1999年03期

5 武會娟;成軍;羅軍;張麗娟;;順式作用元件的預(yù)測[J];醫(yī)學(xué)分子生物學(xué)雜志;2006年03期

6 王曉亮;顧憲紅;鄭冬梅;;雞小腸上皮細胞的分離及體外培養(yǎng)研究進展[J];中國畜牧獸醫(yī);2010年09期

7 李艷;彭春燕;梁榕旺;趙國琦;金曉君;;雞小腸上皮細胞的分離培養(yǎng)與鑒定[J];中國畜牧獸醫(yī);2011年02期

8 楊文平;高峰;許輝堂;施傳信;趙國剛;周光宏;;雞腸上皮細胞體外原代培養(yǎng)研究[J];江西農(nóng)業(yè)學(xué)報;2007年05期

9 裴沛;熊慧;崔鵬;關(guān)偉軍;馬月輝;;雞小腸上皮干細胞的分離培養(yǎng)[J];華北農(nóng)學(xué)報;2014年05期

10 張未麗;劉智皓;吳風(fēng)瑞;黃寶鋒;韓飛;孫運侶;王德壽;;轉(zhuǎn)錄因子GATA-2在脊椎動物發(fā)育過程中的作用[J];四川動物;2009年06期

相關(guān)碩士學(xué)位論文 前2條

1 蔣易楠;GATA2與Smad4相互作用對TGFβ信號通路負調(diào)控的初步研究[D];安徽醫(yī)科大學(xué);2011年

2 王海霞;山羊小腸上皮細胞系的建立及其對氮素的利用研究[D];揚州大學(xué);2008年

，

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