【摘要】：本研究對內(nèi)蒙古絨山羊與薩能奶山羊的四個時期的皮膚樣品進行了轉(zhuǎn)錄組測序。利用生物信息學分析方法對相同時期的絨山羊與奶山羊的差異表達基因和絨山羊不同時期之間的差異表達基因進行了分析,以期能夠篩選出與次級毛囊周期性生長相關(guān)的候選基因,為深入探索絨毛生長發(fā)育機理提供理論依據(jù)。主要研究結(jié)果如下：(1)利用Illumina Hiseq2000對絨山羊與奶山羊四個時期的皮膚樣品進行了RNA-Seq測序。測序共獲得38G數(shù)據(jù),平均每個樣品4.7G。每個樣品的原始reads均達到3100萬以上,Clean比例在77%-89%以上。檢出的山羊基因總數(shù)為20780,檢出率達到97.15%。(2)分析差異表達基因發(fā)現(xiàn),絨山羊與奶山羊間相同時期共有2409個差異表達基因,其中3月份共有350個差異表達基因,6月份共有220個差異表達基因,9月份共有427個差異表達基因,12月份共有1412個差異表達基因。(3)絨山羊不同時期差異表達基因共550個,其中6月Vs 3月有146個差異表達基因,9月Vs6月有98個差異表達基因,12月Vs9月有83個差異表達基因,3月Vs 12月有223個差異表達基因。(4)GO功能注釋分類顯示,差異表達基因主要富集在細胞、細胞部分、細胞器部分,通過結(jié)合、催化功能參與生物調(diào)控、細胞過程、代謝過程等生物過程。(5)KEGG pathway分析發(fā)現(xiàn),絨山羊與奶山羊差異表達基因有39個富集在與毛囊生長相關(guān)的WNT、SHH、TGFβ、TNF、MAPK和Notch信號通路中。絨山羊不同時期差異基因有10個富集在與毛囊生長相關(guān)的WNT、SHH、TGFβ、TNF和MAPK信號通路中,可作為次級毛囊周期性生長相關(guān)候選基因,進行后續(xù)功能研究。(6)采用實時定量PCR對10個差異表達進行基因表達水平驗證,發(fā)現(xiàn)實時定量與轉(zhuǎn)錄組分析結(jié)果基本一致,表明測序結(jié)果準確可靠。
[Abstract]:In this study, the skin samples of four periods of Inner Mongolia cashmere goat and sanen goat were sequenced. The differential expression genes between cashmere goats and dairy goats at the same time were analyzed by bioinformatics analysis, and the differential expression genes between cashmere goats and different periods were analyzed in order to be able to screen the secondary hair follicles. The candidate genes related to cyclical growth provide a theoretical basis for exploring the mechanism of villus growth and development. The main results are as follows: (1) RNA-Seq sequencing of skin samples of cashmere goats and dairy goats by Illumina Hiseq2000 was carried out with 38G data, and the original reads of each sample of each sample was obtained by sequencing, and the original reads of each sample of each sample was RNA-Seq. The total number of Clean and Clean was above 77%-89%. The total number of goat genes was 20780, and the detection rate was 97.15%. (2). There were 2409 differentially expressed genes in the same period between cashmere goats and dairy goats. There were 350 differentially expressed genes in March, and 220 differentially expressed genes in June, September. There were 427 differentially expressed genes in December, and there were 1412 differentially expressed genes in December. (3) there were 550 differentially expressed genes in cashmere goats, including 146 differentially expressed genes in June Vs March, 98 differentially expressed genes in September months in September, 83 differentially expressed genes in December month December, and 223 differentially expressed genes in March in March and Vs December. (4) GO function (4). The annotated classification showed that the differentially expressed genes were mainly enriched in cells, cell parts and organelles, and were involved in biological processes such as biological regulation, cell process and metabolic process through combination and catalytic function. (5) KEGG pathway analysis found that the differentially expressed genes of cashmere goats and milk goats were enriched in 39 of WNT, SHH, TGF beta, TNF, which were associated with hair follicle growth. In MAPK and Notch signaling pathways, there are 10 different genes in cashmere goats enriched in WNT, SHH, TGF beta, TNF and MAPK signaling pathways related to hair follicle growth, which can be used as candidate genes for periodic growth of secondary follicles and carry out subsequent functional studies. (6) validation of 10 differential expressions by real-time PCR. It was found that the results of real-time quantitative analysis and transcriptome analysis were basically consistent, indicating that the sequencing results were accurate and reliable.
【學位授予單位】：內(nèi)蒙古農(nóng)業(yè)大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：S827

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