本文選題：山羊 + 胚胎　； 參考：《西北農(nóng)林科技大學》2017年碩士論文

【摘要】：轉座(Transposition)作為一種普遍的生物現(xiàn)象,意為將某些特定的遺傳因子從一個基因位點復制/剪切、粘貼至另一個基因位點,而這些能夠自主復制和移動的遺傳因子則稱為轉座子(Tansposable Elements,TE)。按照轉座模式將其分為兩大類:一類是DNA轉座子,以DNA為媒介進行轉座;另一類則是反轉錄轉座子(retrotransposon),又稱逆轉錄元件,通過RNA進行轉座。前者在哺乳動物中存在極少,后者則已被證實在人、小鼠、牛等哺乳動物的早期胚胎發(fā)育、胎盤發(fā)育等多項生理過程中發(fā)揮重要的調控作用。內(nèi)源性逆轉錄病毒元件ERVs(Endogenous Retrovirous)作為一類重要的反轉錄轉座子,它們的整合、表達更是對早期胚胎發(fā)育起著重要的調控作用,本實驗旨在探究山羊基因組中對其早期胚胎發(fā)育的合子激活過程中有調控作用的ERV元件,實驗內(nèi)容主要包括以下幾部分:1.對山羊全基因組分析,通過Reaptmasker、muscl等軟件,挑選出600余條ERV序列,構建ERV數(shù)據(jù)庫。對它們進行一級、二級分類,分析它們的結構,計算各類ERV所占比例并將它們定位至染色體上。2.收集山羊卵母細胞,進行體外受精(IVF)處理,發(fā)育至2-Cell/8-Cell期,將IVF 2-cell期、IVF 8-cell期進行單細胞測序,建立轉錄組數(shù)據(jù)庫,包括LncRNA和mRNA兩部分。3.將兩組數(shù)據(jù)進行雙向blast分析,對IVF 8-cell期特異性高表達而在IVF2-cell期低表達的源于ERVs的轉錄序列,進行QRT-PCR檢測,以此篩選在8-cell期高表達的源于ERVs序列的LncRNA/m RNA,通過顯微注射siRNA進行選定序列的干擾,最終得到兩條在早期胚胎發(fā)育過程中有作用的序列l(wèi)ncRNA TCONS_00460156和mRNA hsd17b11。
[Abstract]:Transposition) as a universal biological phenomenon, it means to copy / cut and paste certain genetic factors from one gene site to another, and these genetic factors which can replicate and move independently are called transposable elements. The transposon is divided into two categories according to transposon mode: one is DNA transposon with DNA as the medium, the other is retrotransposon, also called retrotransposon, which is transposed through RNA. The former is rare in mammals, while the latter has been proved to play an important role in the early embryonic development, placental development and other physiological processes of mammals such as human, mouse, cattle and so on. As a kind of important retrotransposons, endogenous retrovirus elements ERVs(Endogenous retrovirousplay an important role in regulating early embryonic development. The aim of this study was to explore the ERV elements in the goat genome that regulate the activation of zygotes during the development of the goat genome. The main contents of the experiment include the following parts: 1: 1. Based on the whole genome analysis of goat, more than 600 ERV sequences were selected by software such as Reaptmaskermuscl, and the ERV database was constructed. They were classified at the first and second levels, their structures were analyzed, the proportions of various ERV were calculated and mapped to the chromosomes. Goat oocytes were collected for in vitro fertilization (IVF) and developed to 2-Cell/8-Cell stage. Single cell sequencing was carried out at IVF 2-cell stage and IVF 8-cell phase, and transcriptional database was established, including LncRNA and mRNA. 3. Two sets of data were analyzed by bidirectional blast. The transcriptional sequences derived from ERVs were detected by QRT-PCR, which had high specific expression in IVF 8-cell phase but low expression in IVF2-cell phase. In this way, the highly expressed LncRNA/m RNAs derived from ERVs sequences were screened in the 8-cell phase, and two sequences lncRNA TCONS_00460156 and mRNA hsd17b11 were obtained by microinjection of siRNA to interfere with the selected sequences in the process of early embryonic development.
【學位授予單位】：西北農(nóng)林科技大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：S827

【參考文獻】

相關期刊論文 前1條

1 武元峰;欒洋;;內(nèi)源性逆轉錄病毒生物學功能及與腫瘤的關系[J];生命科學;2014年09期

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本文編號：1882890