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基于Solexa测序的獭兔(穴兔)皮肤表型转录组研究

Solexa-Sequencing Based Transcriptome Study of Plaice Skin Phenotype in Rex Rabbits (Oryctolagus cuniculus).

作者信息

Pan Lei, Liu Yan, Wei Qiang, Xiao Chenwen, Ji Quanan, Bao Guolian, Wu Xinsheng

机构信息

Animal Husbandry and Veterinary Institute, Zhejiang Academy of Agricultural Sciences, Hangzhou, Zhejiang, China; Chemistry and Life Science, Zhejiang Normal University, Jinhua, Zhejiang, China.

Animal Husbandry and Veterinary Institute, Zhejiang Academy of Agricultural Sciences, Hangzhou, Zhejiang, China.

出版信息

PLoS One. 2015 May 8;10(5):e0124583. doi: 10.1371/journal.pone.0124583. eCollection 2015.

Abstract

BACKGROUND

Fur is an important genetically-determined characteristic of domestic rabbits; rabbit furs are of great economic value. We used the Solexa sequencing technology to assess gene expression in skin tissues from full-sib Rex rabbits of different phenotypes in order to explore the molecular mechanisms associated with fur determination.

METHODOLOGY/PRINCIPAL FINDINGS: Transcriptome analysis included de novo assembly, gene function identification, and gene function classification and enrichment. We obtained 74,032,912 and 71,126,891 short reads of 100 nt, which were assembled into 377,618 unique sequences by Trinity strategy (N50=680 nt). Based on BLAST results with known proteins, 50,228 sequences were identified at a cut-off E-value ≥ 10-5. Using Blast to Gene Ontology (GO), Clusters of Orthologous Groups (KOG) and Kyoto Encyclopedia of Genes and Genomes (KEGG), we obtained several genes with important protein functions. A total of 308 differentially expressed genes were obtained by transcriptome analysis of plaice and un-plaice phenotype animals; 209 additional differentially expressed genes were not found in any database. These genes included 49 that were only expressed in plaice skin rabbits. The novel genes may play important roles during skin growth and development. In addition, 99 known differentially expressed genes were assigned to PI3K-Akt signaling, focal adhesion, and ECM-receptor interactin, among others. Growth factors play a role in skin growth and development by regulating these signaling pathways. We confirmed the altered expression levels of seven target genes by qRT-PCR. And chosen a key gene for SNP to found the differentially between plaice and un-plaice phenotypes rabbit.

CONCLUSIONS/SIGNIFICANCE: The rabbit transcriptome profiling data provide new insights in understanding the molecular mechanisms underlying rabbit skin growth and development.

摘要

背景

被毛是家兔重要的遗传决定特征;兔毛具有很高的经济价值。我们使用Solexa测序技术评估不同表型的全同胞獭兔皮肤组织中的基因表达,以探索与被毛决定相关的分子机制。

方法/主要发现:转录组分析包括从头组装、基因功能鉴定以及基因功能分类和富集。我们获得了74,032,912条和71,126,891条100 nt的短读段,通过Trinity策略将其组装成377,618条独特序列(N50 = 680 nt)。基于与已知蛋白质的BLAST结果,在截止E值≥10-5时鉴定出50,228条序列。通过对基因本体论(GO)、直系同源簇(KOG)和京都基因与基因组百科全书(KEGG)进行Blast分析,我们获得了几个具有重要蛋白质功能的基因。通过对有毛和无毛表型动物的转录组分析共获得308个差异表达基因;另外209个差异表达基因在任何数据库中均未找到。这些基因包括49个仅在有毛皮肤兔中表达的基因。这些新基因可能在皮肤生长和发育过程中发挥重要作用。此外,99个已知的差异表达基因被归类到PI3K-Akt信号通路、粘着斑和ECM-受体相互作用等通路中。生长因子通过调节这些信号通路在皮肤生长和发育中发挥作用。我们通过qRT-PCR证实了7个靶基因表达水平的改变。并选择一个关键基因进行SNP分析,以发现有毛和无毛表型兔之间的差异。

结论/意义:兔转录组图谱数据为理解兔皮肤生长和发育的分子机制提供了新的见解。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/43cd/4425669/77debaeaa442/pone.0124583.g001.jpg

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