牛多个组织microRNASolexa测序与生物信息学分析

doi:10.16431/j.cnki.1671-7236.2015.06.013

《中国畜牧兽医》---唯一指定的官方网站 ›› 2015, Vol. 42 ›› Issue (6): 1409-1416.doi: 10.16431/j.cnki.1671-7236.2015.06.013

牛多个组织microRNASolexa测序与生物信息学分析

刘飞^1,2, 杨继业^1,3, 孙志颖^1,3, 胡鑫^2,4, 李娟², 孔令娜¹, 梁瑞圆¹, 宋振宇¹, 张路培^1,2, 陈燕², 高雪², 李俊雅², 孙少华¹

1. 河北农业大学动物科技学院, 保定 071000;
2. 中国农业科学院北京畜牧兽医研究所, 北京 100193;
3. 河北农业大学生命科学学院, 保定 071000;
4. 内蒙古民族大学动物科学技术学院, 通辽 028000

修回日期:2015-03-25 出版日期:2015-06-20 发布日期:2015-07-23
通讯作者: 李俊雅, 孙少华 E-mail:jll@iasaas.net.cn;shaohuasun@sina.com
作者简介:刘飞(1987-),男,河北保定人,硕士,研究方向:动物遗传育种,E-mail:1578577822@qq.com
基金资助:
牛MSCs成肌/成脂分化中差异miRNAs的分离及调控机制(31201782)

Solexa Sequencing and Bioinformatics Analysis of microRNA from Bovine Multiple Tissues

LIU Fei^1,2, YANG Ji-ye^1,3, SUN Zhi-ying^1,3, HU Xin^2,4, LI Juan², KONG Ling-na¹, LIANG Rui-yuan¹, SONG Zhen-yu¹, ZHANG Lu-pei^1,2, CHEN Yan², GAO Xue², LI Jun-ya², SUN Shao-hua¹

1. College of Animal Science and Technology, Agriculture University of Hebei, Baoding 071000, China;
2. Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, China;
3. College of Life Science, Agriculture University of Hebei, Baoding 071000, China;
4. College of Animal Science and Technology, Inner Mongolia University for the Nationalities, Tongliao 028000, China

Revised:2015-03-25 Online:2015-06-20 Published:2015-07-23

摘要/Abstract

摘要： 本研究旨在发掘牛更多的microRNA(miRNA)信息及新的miRNA序列,为进一步研究miRNA的生物学功能奠定理论基础.运用高通量测序技术对来自一头西门塔尔公牛和一头荷斯坦母牛的多个组织的小RNA进行混池测序,对测序结果进行了生物信息学分析.随机选取了一条成熟miRNA:bta-miR-2346-5p和两条新预测的miRNAs:novel_miR-48和novel_miR-86,采用茎环RT-PCR进行验证.共鉴定了604条miRNAs,其中429条为牛的已知miRNAs,175条为新预测的miRNAs.通过茎环RT-PCR发现选取的这3条miRNAs在背最长肌、心肌、肝脏、脾脏、肺脏、肾脏、大肠、小肠中均有表达,证明了高通量测序结果的准确性.本试验得到的数据在一定程度上丰富了牛miRNA的数量和种类,为后续牛或其他物种的miRNA相关生物学研究提供了更加详实的信息.

关键词: 牛; 多组织; microRNA; 高通量测序

Abstract: In order to provide more information for further studies of miRNA's biological function,the identification of microRNA of bovine was studied.A Solexa sequencing of multiple tissues from a Simmental bull and a Holstein cow was conducted,and then bioinformatics analysis was performed.Meanwhile,we randomly selected and validated 3 miRNAs (bta-miR-2346-5p,novel_miR-48,novel_miR-86) by stem-loop RT-PCR.From this data set,we identified 604 miRNAs,including 429 known miRNAs and 175 novel miRNAs.And the stem-loop RT-PCR result showed that the 3 miRNAs were ubiquitously expressed in longissimus dorsi muscle,cardiac muscle,liver,spleen,lung,kidney,large intestine and small intestine.The study enriched the amount and kinds of bovine miRNAs which provided more credible information for the miRNAs biological functions study in various species.

Key words: bovine; multiple tissues; microRNA; Solexa sequencing

中图分类号:

S858.23

刘飞, 杨继业, 孙志颖, 胡鑫, 李娟, 孔令娜, 梁瑞圆, 宋振宇, 张路培, 陈燕, 高雪, 李俊雅, 孙少华. 牛多个组织microRNASolexa测序与生物信息学分析[J]. 《中国畜牧兽医》---唯一指定的官方网站, 2015, 42(6): 1409-1416.

LIU Fei, YANG Ji-ye, SUN Zhi-ying, HU Xin, LI Juan, KONG Ling-na, LIANG Rui-yuan, SONG Zhen-yu, ZHANG Lu-pei, CHEN Yan, GAO Xue, LI Jun-ya, SUN Shao-hua. Solexa Sequencing and Bioinformatics Analysis of microRNA from Bovine Multiple Tissues[J]. , 2015, 42(6): 1409-1416.

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牛多个组织microRNASolexa测序与生物信息学分析

Solexa Sequencing and Bioinformatics Analysis of microRNA from Bovine Multiple Tissues

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