本文選題：家蠅　切入點：MdSirt1　出處：《河北大學(xué)》2016年碩士論文

【摘要】：隨著對Sirtuins家族蛋白研究的不斷深入,關(guān)于其功能的報道也越來越廣泛。通過轉(zhuǎn)錄組和數(shù)字表達(dá)譜分析,我們發(fā)現(xiàn)家蠅在受到細(xì)菌刺激后體內(nèi)Sirt1和Sirt4基因表達(dá)水平發(fā)生顯著變化,提示它們可能參與家蠅免疫應(yīng)答過程。本文基于此,本工作以此為出發(fā)點對家蠅應(yīng)激過程中Sirt1和Sirt4基因表達(dá)的變化規(guī)律及其可能存在的調(diào)控機(jī)制進(jìn)行了研究。主要內(nèi)容包括:1、克隆了家蠅Sirt1和Sirt4基因c DNA序列,分別命名為Md Sirt1和Md Sirt4,并對其進(jìn)行了生物信息學(xué)分析。兩條基因所編碼的蛋白Md SIRT和Md SIRT4各具有一段保守的SIR2保守結(jié)構(gòu)域,兩者氨基酸序列與其它物種中的同源蛋白之間均具有較高的同源性。2、利用實時熒光定量PCR和蛋白免疫印跡方法對Md Sirt1和Md Sirt4在家蠅不同發(fā)育時期、不同組織和應(yīng)激過程中的表達(dá)量變化進(jìn)行了分析。Md Sirt1和Md Sirt4在家蠅受到細(xì)菌刺激時分別上調(diào)和下調(diào),同時檢測到Md SIRT4在蛋白水平也發(fā)生了相似的變化。3、Md Sirt1和Md Sirt4在家蠅受到熱激處理時m RNA表達(dá)量分別上調(diào)和下調(diào),同時檢測到Md SIRT4在蛋白水平也發(fā)生了相似的變化。4、Md Sirt1和Md Sirt4在家蠅受到重金屬影響后m RNA表達(dá)量均上調(diào),同時檢測到Md SIRT4在蛋白水平也發(fā)生了相似的變化。5、在對Md Sirt1干擾后我們發(fā)現(xiàn)家蠅幼蟲應(yīng)對細(xì)菌刺激的能力下降;而對Md Sirt4進(jìn)行了干擾后,我們發(fā)現(xiàn)家蠅核轉(zhuǎn)錄因子Md NF-κB表達(dá)量上升,同時檢測到家蠅抗菌肽的表達(dá)量上調(diào)。
[Abstract]:With the further study of Sirtuins family proteins, more and more reports about its function have been reported.By the analysis of transcriptome and digital expression profile, we found that the expression level of Sirt1 and Sirt4 genes in housefly was significantly changed after bacterial stimulation, suggesting that they may be involved in the immune response of housefly.Based on this work, the changes of Sirt1 and Sirt4 gene expression and their possible regulatory mechanisms during housefly stress were studied.The main contents include: 1, cloned the c DNA sequence of housefly Sirt1 and Sirt4 gene, named Md Sirt1 and Md Sirt4 respectively, and analyzed them by bioinformatics.The proteins Md SIRT and Md SIRT4 encoded by the two genes each have a conserved SIR2 conserved domain.The amino acid sequence of Md Sirt1 and Md Sirt4 in Musca domestica at different developmental stages were analyzed by real-time fluorescence quantitative PCR and Western blot.The changes of expression of Md Sirt1 and Md Sirt4 in different tissues and stress were analyzed. Md Sirt1 and Md Sirt4 were up-regulated and down-regulated respectively when housefly was stimulated by bacteria.It was also found that Md SIRT4 had a similar change in protein level. Md Sirt1 and Md Sirt4 were up-regulated and down-regulated respectively during heat shock treatment of Musca domestica.It was also found that Md SIRT4 had a similar change in protein level. 4Md Sirt1 and Md Sirt4 were up-regulated after heavy metal exposure in Musca domestica.At the same time, Md SIRT4 had a similar change in protein level. 5. After interfering with Md Sirt1, we found that the ability of housefly larvae to respond to bacterial stimulation was decreased, while Md Sirt4 was interfered with.We found that the expression of nuclear transcription factor Md NF- 魏 B was increased and the expression of antimicrobial peptide in housefly was up-regulated.
【學(xué)位授予單位】：河北大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：Q78

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1 顧冀海;家蠅MdSirt1和MdSirt4基因的克隆及功能分析[D];河北大學(xué);2016年

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本文編號：1724429