牦牛卵巢小RNA高通量测序及生物信息学分析

doi:10.11843/j.issn.0366-6964.2016.01.008

摘要/Abstract

摘要：

本研究旨在利用高通量测序技术描绘牦牛卵巢sRNA图谱，为探析牦牛繁殖性能提供基础。以牦牛卵巢组织作为研究对象，构建牦牛sRNA文库，进行高通量测序和生物信息学分析，最后利用RT-qPCR技术验证miRNA在牦牛卵巢组织中的表达。经测序后得到包含12 126 208 条clean reads，其中特异序列为212 757条；比对分析显示，只有7 383 536 (60.89%) 条总序列及80 981(38.06%) 条特异序列能与牦牛基因组匹配。与黄牛相比，牦牛sRNA中有56个表达量上调，其中miR-135a表达上调最显著；有33个表达下调，其中miR-2316表达下调最显著。RT-qPCR验证6个miRNA在牦牛卵巢组织中的表达情况，定量结果和测序结果基本一致。与牦牛EST序列信息比对后共预测出5个新miRNA。本研究成功绘制牦牛卵巢sRNA图谱，同时证实RNA-Seq高通量测序技术在完善sRNA信息及挖掘新miRNA方面的优势，对研究sRNA在牦牛遗传育种以及以牦牛为重要高原动物模型来进行高原适应性和抗逆性等相关领域的研究具有重要意义。

关键词: 牦牛, 卵巢, 高通量测序, 小RNA, 生物信息学

Abstract:

The small RNA transcriptome profile of yak ovary was described by Solexa deep sequencing，and provided the basic data for future study on yak breeding performance.In this study，yak ovaries were used for building small RNAs library，and then Solexa sequencing and bioinformatics were applied to analysis.Meanwhile，the expression of selected miRNAs was validated by RT-qPCR.After Solexa sequencing，a total of 12 126 208 clean reads were obtained from the ovary library，which contained 212 757 distinct sequences.Alignment analysis showed that 7 383 536 (60.89%) of total sequences and 80 981 (38.06%) distinct sequences were mapped to the yak reference genome.The results of differential expression analysis showed that 56 small RNAs were up-regulated and 33 small RNAs were down-regulated in yak ovary compared to the cattle counterparts.Among these RNA，miR-135a and miR-2316 had the largest fold-change.The expression of 6 selected miRNAs in yak ovary tissues obtained by RT-qPCR was consistent with the Solexa sequencing results.Furthermore，5 novel small RNAs were predicted after compared to the yak EST sequence information.All above suggested the small RNAs transcriptome profile of yak ovary was successfully described，and our results confirmed that Solexa sequencing technology has a great advantage in facilitating small RNA information and mining new miRNAs，and provide valuable small RNA data for further facilitate the yak genetic breeding and study the adaptation and resistance to high altitude and other related fields of highland animal.

Key words: yak, ovary, Solexa sequencing, small RNA, bioinformatics

中图分类号:

S823.8⁺5.2

熊显荣，兰道亮，李键，字向东，林亚秋，马力. 牦牛卵巢小RNA高通量测序及生物信息学分析[J]. 畜牧兽医学报, 2016, 47(1): 55-63.

XIONG Xian-rong，LAN Dao-liang，LI Jian，ZI Xiang-dong，LIN Ya-qiu，MA Li. Solexa Sequencing of Small RNAs in Yak (Bos grunniens) Ovaries and Bioinformatics Analysis[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(1): 55-63.

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