綿羊囊胚和孵化囊胚的轉(zhuǎn)錄組測(cè)序分析
本文選題:綿羊 + 囊胚; 參考:《新疆農(nóng)業(yè)大學(xué)》2015年碩士論文
【摘要】:本文基于RNA-Seq技術(shù),對(duì)綿羊的囊胚和孵化囊胚轉(zhuǎn)錄組進(jìn)行了測(cè)序和分析,獲得了一系列的數(shù)據(jù)資料,并對(duì)數(shù)據(jù)資料進(jìn)行加工整理,得到了主要的研究結(jié)果如下:(1)通過Illumina/Hiseq2000測(cè)序平臺(tái)對(duì)囊胚和孵化囊胚的轉(zhuǎn)錄組進(jìn)行高通量測(cè)序,并對(duì)數(shù)據(jù)進(jìn)行過濾、重組、注釋等,分別得到47664034和47821016條Clean reads,比對(duì)到參考基因組上的reads分別達(dá)到71.48%和72.90%。(2)囊胚和孵化囊胚轉(zhuǎn)錄組分別得到了19318和18288個(gè)轉(zhuǎn)錄本,其中覆蓋度達(dá)到90%-100%的轉(zhuǎn)錄本分別為8135和7823個(gè)。囊胚預(yù)測(cè)到2879個(gè)新轉(zhuǎn)錄本,孵化囊胚預(yù)測(cè)到1277個(gè)新轉(zhuǎn)錄本。(3)差異表達(dá)基因共有7700個(gè),囊胚-VS-孵化囊胚表達(dá)上調(diào)基因3129個(gè),表達(dá)下調(diào)基因4571個(gè)。(4)囊胚和孵化囊胚發(fā)生可變剪切的類型中外顯子跳躍、內(nèi)含子保留、5’端剪切位點(diǎn)替換、3’端剪切位點(diǎn)替換的數(shù)量分別為29117和11332,6864和5698,14899和8190,16321和9431個(gè)。(5)囊胚和孵化囊胚篩選出的SNP位點(diǎn)數(shù)量為97620和67471個(gè),且篩選出的SNP類型中轉(zhuǎn)換類突變明顯要多于顛換類突變。測(cè)序結(jié)果中對(duì)基因的結(jié)構(gòu)優(yōu)化結(jié)果為:囊胚期優(yōu)化基因數(shù)為5641個(gè),孵化囊胚優(yōu)化基因數(shù)5904個(gè)。本研究進(jìn)行的綿羊轉(zhuǎn)錄組測(cè)序分析涉及轉(zhuǎn)錄組的基礎(chǔ)數(shù)據(jù)分析和高級(jí)分析,為綿羊胚胎轉(zhuǎn)錄組的進(jìn)一步研究提供了豐富的基礎(chǔ)數(shù)據(jù)資料,并且為綿羊胚胎發(fā)育過程中基因的調(diào)控提供了一定理論基礎(chǔ),同時(shí)差異表達(dá)基因的篩選和分析反映了基因的表達(dá)情況,可以為相關(guān)生長(zhǎng)過程中分子調(diào)控機(jī)理的深入研究提供基礎(chǔ)。新轉(zhuǎn)錄本預(yù)測(cè)、可變剪切和SNP位點(diǎn)的篩選拓寬了綿羊轉(zhuǎn)錄組的相關(guān)信息,為進(jìn)一步研究綿羊胚胎發(fā)育尤其是囊胚和孵化囊胚在著床前的過程中的基因調(diào)控提供了數(shù)據(jù)參考。
[Abstract]:Based on RNA-Seq technology, the transcriptome of blastocysts and hatched blastocysts of sheep were sequenced and analyzed. A series of data were obtained and processed. The main results are as follows: (1) High-throughput sequencing of blastocysts and hatched blastocysts by Illumina/Hiseq2000 sequencing platform, filtering, recombination, annotation, etc. 47664034 and 47821016 Clean transcripts were obtained respectively, and the reads compared to the reference genome reached 71.48% and 72.90.1%, respectively. 19318 and 18288 transcripts were obtained from blastocyst and hatched blastocyst transcriptome, respectively, of which 8135 and 7823 transcripts with coverage of 90-100% were obtained, respectively. The differentially expressed genes of blastocyst and hatched blastocyst were estimated to be 2879 new transcripts and 1277 new transcripts respectively. There were 7700 differentially expressed genes in blastocyst and 3129 up-regulated genes in blastocyst -VS- hatching blastocyst. The expression of down-regulated gene 4571. T4) the exon jumps in the types of blastocysts and hatched blastocysts with variable shear. The number of SNP site substitutions at the 3'end of the intron reserved 5 'end splicing sites were 29117 and 11332n6864 and 5698899 and 81908116321 and 9431 respectively) the number of SNP loci screened from blastocysts and hatched blastocysts were 97620 and 67471, respectively. Moreover, the transmutation of SNP was more than that of transverse-class mutation. The results of sequencing showed that the number of optimal genes in blastocyst stage was 5 641 and that of hatching blastocyst was 5 904. The analysis of sheep transcriptome sequencing in this study involves basic data analysis and advanced analysis of transcriptome, which provides abundant basic data for further study of sheep embryo transcriptome. It provides a theoretical basis for the regulation of genes during the development of sheep embryos, and the screening and analysis of differentially expressed genes reflect the expression of genes. It can provide the basis for the further study of molecular regulation mechanism in the process of growth. New transcripts predict that the screening of variable shear and SNP sites broadens the information of sheep transcriptome and provides a data reference for further study on the gene regulation of blastocysts and hatched blastocysts in sheep before implantation.
【學(xué)位授予單位】:新疆農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】:碩士
【學(xué)位授予年份】:2015
【分類號(hào)】:S826
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