深度测序鉴定绵羊microRNA转录组

本研究采集的Texel绵羊胎儿的背最长肌样品包括5个发育阶段:70、85、100、120和135 d。通过采用Solexa测序技术对混池样品进行了microRNA(miRNA)深度测序,获得了16532850条原始序列读数。使用ACGT101-miR v4.2分析软件对miRNA测序数据进行了深度发掘;通过与最新的哺乳动物成熟miRNA序列(miRNA数据库:miRBase v17.0)、miRNA前体序列、绵羊基因组序列(绵羊基因组数据库,2010年2月)的比对分析,对绵羊microRNA转录组数据库进行了大幅更新,pre-miRNA(miRNA前体)序列增至1529条,编码的miRNA成熟体序列增至1999条。通过实时荧光定量PCR技术对深度测序获得的5个miRNA在混池样品中进行了验证。绵羊miRNA的研究起步较晚,而miRNA的发现与鉴定将促进基因调控机制及miRNA功能的研究。

Identification of the microRNAome in Texel Sheep by Deep Sequencing

Information concerning the sheep microRNA (miRNA) had been long overdue, the discovery and identification of miRNAs would further promote the study of miRNA functions and gene regulatory mechanisms. Samples were culled that included five developmental stages: The longissimus dorsi muscles of Texel fetuses were at 70, 85, 100, 120 and 135 days. We performed a comprehensive search for Texel miRNA on the pool library by Solexa deep sequencing; then obtained 16532850 raw reads. This study used the ACGT101-miR v4.2 analysis software to explore the sequencing data. The sequencing results were analyzed using the latest mammalian mature miRNA (in miRBase v17.0), precursors (pre-miRNAs), and sheep genome (Ovis aries genome database, Feb 2010). Our results extend the repertoire of sheep microRNAome to 1529 pre-miRNAs encoding for 1999 mature miRNA. We preformed quantitative Real-time PCR experiments for selected 5 miRNA in the pool sample and found agreement between the sequencing and Real-time PCR data. These results were prelude to the advancement in sheep biology.