日本七鳃鳗物种特异性microRNAs及其前体识别与验证

doi:10.16288/j.yczz.14-411

遗传 ›› 2015, Vol. 37 ›› Issue (3): 283-291.doi: 10.16288/j.yczz.14-411

日本七鳃鳗物种特异性microRNAs及其前体识别与验证

刘欣^1,2,张洁^1,2,赵春晖^1,2,李铁松^1,2,王继红^1,2,李庆伟^1,2

1. 辽宁师范大学生命科学学院,大连 116081; 2. 辽宁师范大学七鳃鳗研究中心,大连 116081

收稿日期:2014-11-25 修回日期:2015-01-19 出版日期:2015-03-20 发布日期:2015-02-10
通讯作者: 李庆伟,教授,博士生导师,研究方向：细胞生物学。E-mail: liqw@263.net E-mail:liuxin@lnnu.edu.cn
作者简介:刘欣,教授,硕士生导师,研究方向：细胞生物学。E-mail: liuxin@lnnu.edu.cn
基金资助:
国家重大基础研究发展规划项目(973计划) (编号：2013CB835304)和国家自然科学基金项目(编号：31271323)资助

The identification and verification of species-specific microRNAs and their precursors in Lampetra japonica

Xin Liu^{1, 2}, Jie Zhang^{1, 2}, Chunhui Zhao^{1, 2}, Tiesong Li^{1, 2}, Jihong Wang^{1, 2}, Qingwei Li^{1, 2}

1. School of Life Science, Liaoning Normal Uuniversity, Dalian 116081, China;
2. Lamprey Research Center, Liaoning Normal University, Dalian 116081, China

Received:2014-11-25 Revised:2015-01-19 Online:2015-03-20 Published:2015-02-10

摘要/Abstract

摘要： MicroRNAs(miRNAs)对参与多种生物代谢过程的基因在转录及转录后水平进行负调控。近年来,随着深度测序及芯片技术的应用,有关miRNA的发现和功能分析在植物和动物中得到广泛研究。文章利用第二代测序技术对日本七鳃鳗(Lampetra japonica)白细胞的小RNA进行了高通量测序,共得到5 207 787条小RNA序列,其中4 739 346条序列可以拼接为10 989种miRNA变体。基于序列相似性分析,发现这10 989个变体序列与306个已知的保守miRNA家族成员序列相匹配;其中,6个保守miRNA家族成员呈极高丰度表达,表明miRNA在物种间具有保守性。70个未注释序列被预测为新的miRNA。通过miRNA微阵列技术鉴定与验证了34个新预测的miRNA在免疫处理的日本七鳃鳗白细胞中表达,其中16个miRNA前体的最低折叠自由能系数大于0.85,说明日本七鳃鳗存在特异性miRNA。这些物种特异性miRNAs的存在可能在日本七鳃鳗的白细胞生长、发育和对疾病的反应中发挥重要的调控作用。

关键词: 日本七鳃鳗, 物种特异性miRNA, 高通量测序, 基因芯片

Abstract: MicroRNAs (miRNAs) negatively regulate genes which are involved in various biological processes of metabolism at both transcriptional and post-transcriptional levels. In recent years, the existence and function of miRNAs have been extensively studied in plants and animals with the application of deep sequencing and microarray technology. In this study, small RNAs from leucocytes of Lampetra japonica (L. japonica) were sequenced using the second generation high-throughput sequencing technology. A total of 5 207 787 small RNA sequences were identified, and 4 739 346 of them assembled into 10 989 variants. Based on sequence similarity analysis, the sequences of these variants matched known miRNAs of 306 conserved families, among which 6 conserved miRNA family members expressed at an extremely high level which reflected the conservatism of miRNAs among species. In addition, 70 unannotated sequences were predicted to be new miRNAs, and 34 of them were further verified expressing in antigen-treated L. japonica leucocytes by miRNA microarray assay. Moreover, the minimal folding free energy indexes for 16 of the 34 miRNA precursors exceed 0.85, indicating the existence of species-specific miRNAs in L. japonica which may play important roles in regulating, growth, development and disease response of L. japonica leukocytes.

Key words: Lampetra japonica, species-specific miRNA, high-throughput sequencing, microarray assay

刘欣, 张洁, 赵春晖, 李铁松, 王继红, 李庆伟. 日本七鳃鳗物种特异性microRNAs及其前体识别与验证[J]. 遗传, 2015, 37(3): 283-291.

Xin Liu, Jie Zhang, Chunhui Zhao, Tiesong Li, Jihong Wang, Qingwei Li. The identification and verification of species-specific microRNAs and their precursors in Lampetra japonica[J]. HEREDITAS(Beijing), 2015, 37(3): 283-291.

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日本七鳃鳗物种特异性microRNAs及其前体识别与验证

The identification and verification of species-specific microRNAs and their precursors in Lampetra japonica

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