摘要
为了筛选不同被毛纤维直径的细毛羊皮肤组织中的差异表达基因,本研究利用Agilent公司绵羊表达谱芯片和Affymetrix公司牛表达谱芯片对不同被毛纤维直径的细毛羊皮肤组织mRNA进行了芯片杂交;运用Sig-nificance Analysis of Microarrays(SAM)法对基因表达谱进行了差异分析;并通过分子注释系统平台(MAS3.0)对差异表达基因进行了功能富集类分析和调控通路分析;通过实时定量PCR对部分差异表达基因进行了验证。芯片试验共检测到267个差异表达的基因,其中上调基因106个,下调基因161个。Kegg和GO分析表明,参与细胞凋亡、增殖及分化、蛋白质代谢、Wnt信号通路以及羊毛角蛋白结构等生物学过程的多个基因表达水平发生了显著变化。所验证的差异表达基因的荧光定量PCR结果与芯片结果基本一致。结果表明,不同被毛纤维直径的细毛羊皮肤组织的基因表达谱存在差别,这些基因的不同表达可能通过改变皮肤毛囊和羊毛分化过程影响被毛纤维直径。这些基因的发现将为进一步筛选影响羊毛纤维直径性状的重要候选基因/分子标记提供研究基础。
The study aimed to screen the differentially expressed genes of skin tissue in fine wool sheep with different fiber diameter. Affymetrix GeneChip ？ Bovine Genome Array and Agilent Sheep Oligo Microarray were used to screen differentially expressed genes of skin tissue in fine wool sheep with different fiber diameter. Significance Analysis of Microarray （SAM） methods was used to identify the differentially expressed genes. GO （Gene Ontology） and the pathway analyses were conducted on differentially expressed genes using a free web-based Molecular Annotation System 3.0 （MAS 3.0）. A total of 267 genes were identified to be differentially expressed genes from 2 different Microarray, 106 were up-regulated expression genes and 161 were down-regulated expression genes. GO and Kegg analysis revealed that the expression levels of the most differentially expressed genes were related to cell proliferation,differentiation,apoptosis, protein biosynthesis, Wnt signaling pathway and Keratin constitution etc. Subsequently, Real-time PCR was performed to validate seven differentially expressed genes which were screened out by the microarray approach and sufficient consistency was observed by the two methods. The results suggest that the gene expression profiles of fine wool sheep with different fiber diameter are variant,their different expression may alter the formation and differentiation of skin follicle and wool fiber, and some important genes identified may be useful in further study on molecu1ar markers of fiber diameter in fine wool sheep.
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2013年第5期681-689,共9页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
国家科技支撑项目(2011BAD28B05)
国家绒毛用羊产业技术体系(CARS-40)
新疆重大专项(200731132-7)
关键词
细毛羊
纤维直径
基因表达
基因芯片
fine wool sheep
fiber diameter
gene expression
microarray
作者简介
狄江（1969-），男，新疆巴里坤人，研究员，博士，主要从事家畜遗传育种领域研究，Tel：0991—4835204，E—mail：dijiang69@163．com
