發(fā)布時間：2018-03-10 22:09

  本文選題：豬　切入點：表觀遺傳學(xué)　出處：《華中農(nóng)業(yè)大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

【摘要】：表觀遺傳學(xué)又稱“擬遺傳學(xué)”、“表遺傳學(xué)”、“外遺傳學(xué)”、“后遺傳學(xué)”,是軟遺傳的重要組成部分。雖然表觀遺傳涉及多個內(nèi)容,但DNA甲基化卻是表觀遺傳研究的最核心、最主要的內(nèi)容之一。DNMT1是一種甲基化轉(zhuǎn)移酶,在細胞中主要承擔(dān)DNA甲基化維持的生物學(xué)功能。目前,在人、小鼠、擬南芥等動植物中,對DNMT1基因開展了大量的研究,得到了一系列豐碩的結(jié)果。不過,這研究主要集中在模式動植物,在家畜中的研究則不夠深入,而系統(tǒng)地從免疫角度探討DNMT1基因功能的研究則更加不足。DNMT1基因參與了機體內(nèi)絕大多數(shù)生物學(xué)過程的調(diào)控,與免疫調(diào)控也有密切的關(guān)系。鑒于此,本研究主要以豬為研究對象,以免疫為重點視角,擬對豬的DNMT1基因開展初步的研究,分別從系統(tǒng)發(fā)育樹、可變剪切體、偶聯(lián)轉(zhuǎn)錄組等角度,對豬DNMT1基因的系統(tǒng)發(fā)生關(guān)系,該基因在鼻粘膜、腦淋巴結(jié)、骨髓、胸腺、脾等免疫組織中的可變剪切情況,以及基于外周血細胞模型的DNMT1基因的偶聯(lián)轉(zhuǎn)錄組開展了較為系統(tǒng)的研究,初步得到研究結(jié)果如下:1.用mega3.1構(gòu)建DNMT1基因在多個物種間的系統(tǒng)發(fā)生樹,從進化角度揭示了豬DNMT1基因與其他物種DNMT1基因的分化關(guān)系,發(fā)現(xiàn)DNMT1基因在各物種間的系統(tǒng)遺傳距離各不相同,但在家養(yǎng)農(nóng)業(yè)動物之間具有更高的保守性。2.利用RT-PCR技術(shù),鑒定了豬DNMT1基因的可變剪切體DNMTo1和DNMTo2,并系統(tǒng)地檢測了鼻粘膜、腦淋巴結(jié)、骨髓、胸腺、脾等免疫組織及對照組織中的DNMT1基因m RNA及其可變剪切體的表達情況,結(jié)果發(fā)現(xiàn)Dnmt1s、Dnmt1o1、Dnmt1o2在各個免疫組織中均有表達,但其豐度和相互間比例在各免疫組織間呈現(xiàn)出差異。3.以豬外周血細胞為模型,利用豬Affymetrix表達譜芯片建立了DNMT1基因的偶聯(lián)轉(zhuǎn)錄組(coupling transcriptome)的分析框架,包括基于DNMT1基因?qū)��?yīng)探針組(probesets)的分布檢測、質(zhì)量控制分析、差異表達基因鑒定、聚類分析、GO注釋等。4.以探針組Ssc.24344.1.S1_a_at表達值的兩尾為基礎(chǔ),鑒定出差異表達轉(zhuǎn)錄本共161個;以探針組Ssc.24344.1.S1_at表達值為協(xié)變量,鑒定出差異表達轉(zhuǎn)錄本135個。5.利用降維策略初步探討了DNMT1基因表達變異對基因組表達模式的影響。我們采用非負矩陣因子化方法對經(jīng)過非特異性過濾后的表達矩陣進行降維處理,構(gòu)建出10個新的因子化“性狀”,并估計了DNMT1基因與各因子之間的相關(guān)系數(shù)與回歸系數(shù),從宏觀上揭示了DNMT1基因的表達變異與基因組的整體表達模式相偶聯(lián)。本研究以免疫生物學(xué)為視角,從多個角度對豬DNMT1基因開展了研究,得到了多個有意義的研究結(jié)果,積累了豬DNMT1基因的基礎(chǔ)數(shù)據(jù),為進一步深入研究豬DNMT1基因的免疫調(diào)控功能奠定了數(shù)據(jù)基礎(chǔ)。
[Abstract]:Epigenetics, also known as "pseudogenetics", "epigenetics", "extrinsic genetics" and "post-genetics", is an important part of soft genetics. Although epigenetics involves many aspects, DNA methylation is the core of epigenetic research. DNMT1 is a kind of methylation transferase, which is mainly responsible for the biological function of DNA methylation in cells. At present, DNMT1 gene has been widely studied in animals and plants, such as human, mouse, Arabidopsis thaliana and so on. A series of fruitful results have been obtained. However, this study is mainly focused on model animals and plants, but not enough on domestic animals. However, the systematic study of the function of DNMT1 gene from the immune angle is even less. DNMT1 gene is involved in the regulation of most biological processes in the organism and has close relationship with immune regulation. In view of this, the study mainly focuses on pigs. From the point of view of immunity, this paper intends to carry out a preliminary study on the DNMT1 gene of pigs. From the perspectives of phylogenetic tree, variable shearing body and coupling transcriptome, the phylogenetic relationship of porcine DNMT1 gene was studied. The gene was found in nasal mucosa, brain lymph node, bone marrow, and so on. The variable shearing of thymus and spleen and the coupling transcriptome of DNMT1 gene based on peripheral blood cell model have been studied systematically. The results are as follows: 1. The phylogenetic tree of DNMT1 gene was constructed with mega3.1, and the relationship between porcine DNMT1 gene and DNMT1 gene of other species was revealed from the perspective of evolution. It was found that the phylogenetic distance of DNMT1 gene varied from species to species, but there was a higher conservation among farm animals raised at home. Using RT-PCR technique, the variable shearing bodies DNMTo1 and DNMTo2 of porcine DNMT1 gene were identified, and the nasal mucosa was systematically detected. The expression of DNMT1 gene m RNA and its variable shearing bodies in brain lymph nodes, bone marrow, thymus, spleen and control tissues were observed. The results showed that Dnmt1snmt1o1, Dnmt1o1 and Dnmt1o2 were expressed in all immune tissues. However, the abundance and proportion of DNMT1 gene were different among different immune tissues. Using porcine peripheral blood cells as a model, the analysis framework of coupling transcriptome of DNMT1 gene was established by using porcine Affymetrix expression microarray. Including distribution detection, quality control analysis, differential expression gene identification, cluster analysis and so on. Based on the two ends of Ssc.24344.1.S1_a_at expression value of probe group, 161 differentially expressed transcripts were identified. The Ssc.24344.1.S1_at expression value of probe group was used as covariable. 135 differentially expressed transcripts were identified. The effects of DNMT1 gene expression variation on genomic expression patterns were preliminarily discussed by dimensionality reduction strategy. We used non-negative matrix factorization method to analyze the expression moments after non-specific filtering. The array is processed by dimensionality reduction, Ten new factorization "traits" were constructed, and the correlation coefficient and regression coefficient between DNMT1 gene and each factor were estimated. It is revealed that the expression variation of DNMT1 gene is coupled with the overall expression pattern of the genome. From the perspective of immunobiology, the porcine DNMT1 gene was studied from several angles, and many meaningful results were obtained. The basic data of porcine DNMT1 gene were accumulated, which laid a data foundation for further study on the immunological regulation function of porcine DNMT1 gene.
【學(xué)位授予單位】：華中農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S828

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本文編號：1595282