家兔肌肉组织miRNA Solexa测序与生物信息学分析

张翔宇 邝良德 李丛艳 张翠霞 郑洁 杨超 任永军 郭志强 唐丽 邓晓东 雷岷 黄邓萍 谢晓红

摘要：miRNA是一类长度约22 bp的小分子非编码RNA，在骨骼肌生长和肌细胞分化过程中起着重要的调控作用。利用Solexa测序技术对3只84日龄成年公兔的腿肌进行了miRNA测序，旨在评估高通量测序在发掘家兔miRNA中的数据质量以及解析家兔肌肉组织中miRNA的生物学特征。结果表明，除前4位碱基的错误率略高于0.5%外，其他位置的碱基错误率均低于0.5%，说明测序质量好、可信度高，获得的序列可用于后续miRNA的鉴定分析。纯净序列中其他类型RNA的比例仅占2.07%。肉兔肌肉中18～24 nt的小RNA首位碱基以U为主，其次为碱基A，具有很强的偏向性。miRNA序列长度主要分布在20～24 nt范围内，占纯净序列的90%以上。miRNA长度分布的不平衡和首位碱基的偏向性可能与miRNA的作用机理和具体生物学功能有关。

关键词：家兔；肌肉；高通量测序；miRNA

中图分类号：S829.1 文献标识码：A

文章编号：0439-8114（2018）16-0097-05

DOI：10.14088/j.cnki.issn0439-8114.2018.16.023 开放科学（资源服务）标识码（OSID）：

Solexa Sequencing and Bioinformatics Analysis on miRNA from Rabbit Muscle

ZHANG Xiang-yu，KUANG Liang-de，LI Cong-yan，ZHANG Cui-xia，ZHENG Jie，YANG Chao，REN Yong-jun，

GUO Zhi-qiang，TANG Li，DENG Xiao-dong，LEI Min，HUANG Deng-ping，XIE Xiao-hong

（Sichuan Animal Science Academy/Key Laboratory of Animal Genetics and Breeding in Sichuan Province，Chengdu 610066， China）

Abstract： miRNA as a kind of micromolecule non-coding RNA with the length of about 22 bp plays an important regulatory role in skeletal muscle growth and muscle cell differentiation. This research uses Solexa sequencing technology to conduct miRNA sequencing on three eight-four-day-old adult male rabbits， aiming at evaluating the quality of high-throughput sequencing in meat-rabbit miRNA data mining， and exploring the biological characteristics of miRNA in meat-rabbit muscular tissues. Results show that， except for bases in first three positions with an error rate slightly higher than 0.5%， bases in other positions have an error rate of lower than 0.5%， which indicates good sequencing quality， high reliability， and the applicability of obtained sequence in subsequent miRNA identification and analysis. Other types of RNA in pure sequences only take up 2.07%. The 18～24 nt small RNA first base in meat-rabbit muscle is mainly U base， and A base for the second with strong directivity. miRNA sequence length is mainly within the range of 20～24 nt， which takes up more than 90% of pure sequences. miRNA sequence length imbalance and the directivity of first-place base may be related to the action mechanism and specific functions of miRNA.

Key words： rabbit； muscle； high throughput sequencing； miRNA

家兔是節粮型食草小家畜，也是重要的人类肉食来源之一，还是许多人类疾病研究的理想动物模型。过去30年间，世界各国对家兔生长速度和肉质的遗传改良，已获得较大研究进展。欧洲的兔业发达国家已选育出齐卡（ZIKA）、伊拉（HYLA）和伊普吕（ELCO）等具有优良生长性能和肉用性能的肉兔配套系[1]。随着经济的发展和消费观念的改变，如何进一步提高兔肉产量和品质成为各国肉兔育种公司共同关注的焦点问题。对家兔肌肉生长发育机制认识的不足，尤其是对重要的调控因子及其作用机理知之甚少，极大地限制了家兔产肉性能的进一步提高。……