山羊肌肉组织microRNA Solexa测序与生物信息学分析

畜牧兽医学报

山羊肌肉组织microRNA Solexa测序与生物信息学分析

凌英会^1，2，张晓东^1，2，王丽娟^1，2，殷捷^1，2，向浩^1，2，李运生^1，2，张运海^1，2，张子军^1，2，丁建平^1，2*，章孝荣^1，2*

（1. 安徽农业大学动物科技学院，合肥 230036； 2. 安徽省地方畜禽遗传资源保护和生物育种重点实验室，合肥 230036）

收稿日期:2012-09-28 出版日期:2013-03-23 发布日期:2013-03-25
通讯作者: 章孝荣，教授，博士生导师，主要从事动物遗传育种与繁殖方面研究，Tel：0551-65785928，E-mail：zxr@ahau.edu.cn；丁建平，副教授，硕士生导师，主要从事动物生殖调控方面研究， E-mail：dingjianping@ahau.edu.cn
作者简介:凌英会（1981-），男，安徽安庆人，博士，主要从事动物遗传育种方面研究，E-mail：caaslyh@163.com
基金资助:
国家高技术研究发展计划(863)项目(2011AA100307);安徽省技术创新工程试点省专项 (11Z0101095);安徽省科技计划项目(11010302108)

Solexa Sequencing and Bioinformatics Analysis on MicroRNA from the Goat Muscle

LING Ying-hui^{1, 2}, ZHANG Xiao-dong ^{1, 2}, WANG Li-juan^{1, 2}, YIN Jie^1,2, XIANG Hao^1,2, LI Yun-sheng^1,2, ZHANG Yun-hai ^{1, 2}, ZHANG Zi-jun^1,2, DING Jian-ping^{1, 2*}, ZHANG Xiao-rong^1,2*

(1. College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036,China; 2. Local Animal Genetic Resources Conservation and Biobreeding Laboratory of Anhui Province, Hefei 230036, China)

Received:2012-09-28 Online:2013-03-23 Published:2013-03-25

摘要/Abstract

摘要：

本研究旨在挖掘山羊肌肉组织miRNA，获取山羊肌肉组织miRNA表达谱并分析其生物学特征，为进一步研究特定miRNA参与山羊肌肉细胞的增殖与分化等生长发育机制奠定理论基础。从山羊肌肉组织中提取总RNA，分离、构建小片段RNA文库，利用Solexa方法进行测序并进行生物信息学分析，最后利用q-PCR技术验证miRNA在山羊肌肉组织中的表达情况。共鉴定出517个物种间保守的和2个山羊基因组特有的miRNA。其中306个表达量在100以上。随机选择8个miRNA，利用q-PCR验证其在山羊肌肉组织中的表达情况，结果与测序结果一致。同时miRNA靶基因预测和功能分析表明，很多miRNA可能影响一些与动物肌肉发育相关的细胞信号通路。测序成功获取了山羊肌肉组织miRNA序列及其表达谱，山羊肌肉组织miRNA表达丰富且表达量各异。本研究也为进一步研究miRNA调控哺乳动物肌肉发育奠定了理论基础。

关键词: 山羊, 肌肉, microRNA, 测序, 特征

Abstract:

Characterization of microRNA expression in goat muscle was studied, with the purposes of providing basic information for further studies of goat growth and development activities such as goat muscle cell proliferation and differentiation. The small RNAs isolated from total RNA of goat muscle were sequenced by Solexa and then bioinformatics analysis was perfomed. Meanwhile, the expression of selected miRNA was validated by q-PCR. Based on Solexa sequencing and bioinformatics analysis, 517 muscle miRNAs, which are conservative on evolution of mammals (sheep, bovine, swine, horse, dog), and 2 goat genome-specific miRNA were identified. Among them, 306 exceeded 100 in the expression levels. The expression of 8 selected miRNA in muscle tissues obtained by q-PCR was agreement with Solexa sequencing results. MiRNA target gene prediction and functional analysis showed that many miRNAs among them may affect cell signaling pathways and animal muscle development. Succeed in construction of expression profiling of miRNA which are abundant and differentially expressed in goat muscle tissues. This study provides important information on the role of miRNA regulation in muscle growth and development.

Key words: goat, muscle, microRNA, sequencing, characteristic

中图分类号:

S827.2

凌英会，张晓东，王丽娟，殷捷，向浩，李运生，张运海，张子军，丁建平，章孝荣. 山羊肌肉组织microRNA Solexa测序与生物信息学分析[J]. 畜牧兽医学报.

LING Ying-hui, ZHANG Xiao-dong,WANG Li-juan1, YIN Jie, XIANG Hao, LI Yun-sheng, ZHANG Yun-hai,ZHANG Zi-jun, DING Jian-ping, ZHANG Xiao-rong. Solexa Sequencing and Bioinformatics Analysis on MicroRNA from the Goat Muscle[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA.

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