细毛羊次级毛囊形态发生诱导期转录组差异表达分析

发布时间：2018-05-02 11:47

本文选题：细毛羊 + 次级毛囊　；参考：《甘肃农业大学》2015年硕士论文

【摘要】：次级毛囊决定着细羊毛产量并对其纤维直径具有重要的影响,阐明调控绵羊次级毛囊形态发生的分子机制,找到特异性调控绵羊次级毛囊生长发育的调控因子和功能基因,将为利用分子育种技术改良羊毛品质提供可靠的科学依据。大量研究已经证实LncRNA对器官形成、细胞分化和疾病发生等各种生理和病理过程都有重要的调控作用,但LncRNA对毛囊形态发生中的调控作用机制还知之甚少。因此本研究应用基于高通量的链特异RNA-seq技术比较次级毛囊形态发生不同阶段(形态发生前期-87d、基板期-96d)的皮肤转录组特征,将得到的Clean reads与绵羊参考基因组(Sheep Genome v3.1)进行比对。主要研究结果如下:(1)次级毛囊形态发生中主要RNA类型为mRNA,分别占Reads的61.92%、62.51%,未知的、潜在为LncRNA的Reads占36.41%、35.08%。用cufflinks筛选出了1288个候选LncRNA转录本,然后用CNCI分析、Pfam蛋白结构域分析、phyloCSF分析剔除具有编码潜能的候选转录本,共获得LncRNA 884个,其中LincRNA 622个、intronic LncRNA188个、anti-sense LncRNA74个。(2)用EdgeR进行差异基因、LncRNA表达分析,显著差异基因192个,其中上调67个,下调125个;显著差异LncRNA 15个,其中上调6个、下调9个。选取10个差异基因、LncRNA进行链特异RT-PCR验证,结果与测序结果基本吻合。(3)通过GO、KEGG等对差异表达基因进行功能富集类分析和调控通路分析,筛选得到的差异基因、LncRNA分别被富集到1023、191个GO term,136、12个Pathway。显著富集的GO term分别157、32个,主要为甲酸脱氢酶复合物、转录调节、DNA依赖、阳离子结合等,显著富集的信号通路为PPAR通路(padj0.05)。结果初步表明,在毛囊形态发生不仅受mRNA调控,而且LncRNA也参与其中,这些差异基因、LncRNA的发现将为进一步阐明毛囊形态发生的分子调控机制提供理论依据。
[Abstract]:The secondary hair follicles determine the yield of fine wool and have an important effect on the fiber diameter. The molecular mechanism of regulating the morphogenesis of the secondary hair follicles of sheep is clarified, and the regulatory factors and functional genes that specifically regulate the growth and development of the secondary hair follicles of sheep are found. It will provide reliable scientific basis for using molecular breeding technology to improve wool quality. A large number of studies have confirmed that LncRNA plays an important role in various physiological and pathological processes, such as organ formation, cell differentiation and disease development, but little is known about the regulatory mechanism of LncRNA on hair follicle morphogenesis. Therefore, a high-throughput strand specific RNA-seq technique was used to compare the skin transcriptional characteristics of secondary hair follicle morphogenesis at different stages (premorphogenetic stage -87 d, substrate stage -96 d). The obtained Clean reads was compared with sheep reference genome sheet Genome v3.1. The main results were as follows: (1) the main type of RNA in secondary hair follicle morphogenesis was mRNAs, which accounted for 61.92% of Reads and 62.51% of Reads, respectively, and 36.41% of Reads with unknown potential for LncRNA accounted for 35.08%. 1288 candidate LncRNA transcripts were screened by cufflinks, and 884 candidate transcripts with encoding potential were eliminated by CNCI analysis of Pfam protein domain. Among them, LincRNA 622 intronic LncRNA188 (anti-sense LncRNA74. 2) EdgeR was used to analyze the expression of differentially expressed LncRNAs. There were 192 significant differentially expressed genes, including 67 up-regulated genes and 125 down-regulated genes, and 15 significantly different LncRNA genes, including 6 up-regulated genes and 9 down-regulated genes. Ten differentially expressed genes (LncRNAs) were selected for RT-PCR validation. The results were in good agreement with the sequencing results. LncRNA was enriched to 1023191 go terms136and 12 Pathways, respectively. The significant enrichment of go term was 157,32, mainly formate dehydrogenase complex, transcriptional regulatory DNA-dependent, cationic binding, etc. The significantly enriched signal pathway was PPAR pathway padj 0.05. The results showed that hair follicle morphogenesis was not only regulated by mRNA, but also LncRNA was involved. The discovery of these differentially expressed genes, LncRNA, will provide a theoretical basis for further elucidating the molecular regulation mechanism of hair follicle morphogenesis.
【学位授予单位】：甘肃农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S826

【共引文献】