山羊基因组中源于ERVs的lncRNA在早期胚胎发育中的作用

发布时间：2018-05-13 11:05

本文选题：山羊 + 胚胎　；参考：《西北农林科技大学》2017年硕士论文

【摘要】：转座(Transposition)作为一种普遍的生物现象,意为将某些特定的遗传因子从一个基因位点复制/剪切、粘贴至另一个基因位点,而这些能够自主复制和移动的遗传因子则称为转座子(Tansposable Elements,TE)。按照转座模式将其分为两大类:一类是DNA转座子,以DNA为媒介进行转座;另一类则是反转录转座子(retrotransposon),又称逆转录元件,通过RNA进行转座。前者在哺乳动物中存在极少,后者则已被证实在人、小鼠、牛等哺乳动物的早期胚胎发育、胎盘发育等多项生理过程中发挥重要的调控作用。内源性逆转录病毒元件ERVs(Endogenous Retrovirous)作为一类重要的反转录转座子,它们的整合、表达更是对早期胚胎发育起着重要的调控作用,本实验旨在探究山羊基因组中对其早期胚胎发育的合子激活过程中有调控作用的ERV元件,实验内容主要包括以下几部分:1.对山羊全基因组分析,通过Reaptmasker、muscl等软件,挑选出600余条ERV序列,构建ERV数据库。对它们进行一级、二级分类,分析它们的结构,计算各类ERV所占比例并将它们定位至染色体上。2.收集山羊卵母细胞,进行体外受精(IVF)处理,发育至2-Cell/8-Cell期,将IVF 2-cell期、IVF 8-cell期进行单细胞测序,建立转录组数据库,包括LncRNA和mRNA两部分。3.将两组数据进行双向blast分析,对IVF 8-cell期特异性高表达而在IVF2-cell期低表达的源于ERVs的转录序列,进行QRT-PCR检测,以此筛选在8-cell期高表达的源于ERVs序列的LncRNA/m RNA,通过显微注射siRNA进行选定序列的干扰,最终得到两条在早期胚胎发育过程中有作用的序列lncRNA 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.
【学位授予单位】：西北农林科技大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S827

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