绒山羊次级毛囊周期性生长相关基因的筛选

发布时间：2018-08-03 11:48

【摘要】：本研究对内蒙古绒山羊与萨能奶山羊的四个时期的皮肤样品进行了转录组测序。利用生物信息学分析方法对相同时期的绒山羊与奶山羊的差异表达基因和绒山羊不同时期之间的差异表达基因进行了分析,以期能够筛选出与次级毛囊周期性生长相关的候选基因,为深入探索绒毛生长发育机理提供理论依据。主要研究结果如下：(1)利用Illumina Hiseq2000对绒山羊与奶山羊四个时期的皮肤样品进行了RNA-Seq测序。测序共获得38G数据,平均每个样品4.7G。每个样品的原始reads均达到3100万以上,Clean比例在77%-89%以上。检出的山羊基因总数为20780,检出率达到97.15%。(2)分析差异表达基因发现,绒山羊与奶山羊间相同时期共有2409个差异表达基因,其中3月份共有350个差异表达基因,6月份共有220个差异表达基因,9月份共有427个差异表达基因,12月份共有1412个差异表达基因。(3)绒山羊不同时期差异表达基因共550个,其中6月Vs 3月有146个差异表达基因,9月Vs6月有98个差异表达基因,12月Vs9月有83个差异表达基因,3月Vs 12月有223个差异表达基因。(4)GO功能注释分类显示,差异表达基因主要富集在细胞、细胞部分、细胞器部分,通过结合、催化功能参与生物调控、细胞过程、代谢过程等生物过程。(5)KEGG pathway分析发现,绒山羊与奶山羊差异表达基因有39个富集在与毛囊生长相关的WNT、SHH、TGFβ、TNF、MAPK和Notch信号通路中。绒山羊不同时期差异基因有10个富集在与毛囊生长相关的WNT、SHH、TGFβ、TNF和MAPK信号通路中,可作为次级毛囊周期性生长相关候选基因,进行后续功能研究。(6)采用实时定量PCR对10个差异表达进行基因表达水平验证,发现实时定量与转录组分析结果基本一致,表明测序结果准确可靠。
[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.
【学位授予单位】：内蒙古农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S827

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