[1]郑月,陈坤琳,李惠侠,等.热应激奶牛血清miRNA表达谱及miR-181a靶基因分析[J].南京农业大学学报,2014,37(6):130-136.[doi:10.7685/j.issn.1000-2030.2014.06.019]
 ZHENG Yue,CHEN Kunlin,LI Huixia,et al.Profiling of differential expression of microRNAs in serum of heat-stresseded cows and bioinformatic analyses of miR-181a target genes[J].Journal of Nanjing Agricultural University,2014,37(6):130-136.[doi:10.7685/j.issn.1000-2030.2014.06.019]
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热应激奶牛血清miRNA表达谱及miR-181a靶基因分析()
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《南京农业大学学报》[ISSN:1000-2030/CN:32-1148/S]

卷:
37卷
期数:
2014年6期
页码:
130-136
栏目:
出版日期:
2014-11-17

文章信息/Info

Title:
Profiling of differential expression of microRNAs in serum of heat-stresseded cows and bioinformatic analyses of miR-181a target genes
作者:
郑月 陈坤琳 李惠侠 王根林
南京农业大学动物科技学院, 江苏 南京 210095
Author(s):
ZHENG Yue CHEN Kunlin LI Huixia WANG Genlin
College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, China
关键词:
荷斯坦奶牛热应激miRNAs靶基因信号通路
Keywords:
Holstein cowsheat-stressedmiRNAstarget genessignal pathway
分类号:
S813.3
DOI:
10.7685/j.issn.1000-2030.2014.06.019
摘要:
通过分析热应激奶牛和正常奶牛血清差异表达miRNA,及对重要的候选miRNA进行靶基因预测及相关生物信息学分析,探索miRNA在荷斯坦奶牛发生热应激过程中的作用及其调控机制。利用miRNA高通量测序技术对奶牛血清中miRNA表达谱进行检测和分析以筛选差异表达miRNA;用RT-qPCR方法验证4个在热应激奶牛血清与正常奶牛血清中显著差异表达的候选miRNAs;采用Targetscan生物信息学计算方法预测目标miR-181a(对应激以及免疫反应可能起重要调节作用)的靶基因,并对靶基因进行GO注释描述、富集分析及KEGG富集。结果表明:miRNA高通量测序分析发现共有52个差异表达极显著miRNA(P<0.01)。候选的4个miRNAs(miR-19a、miR-19b、miR-181a、miR-1246)的RT-qPCR检测结果与测序分析结果具有很好的一致性。GO和KEGG分析结果显示,miR-181a的靶基因与应激反应以及免疫功能密切相关,并且靶基因在B细胞和T细胞受体信号通路中显著富集。结论:在热应激奶牛与正常奶牛血清中存在差异表达的miRNAs,某些重要的miRNA(如miR-181a)可能以独特的通路(如B细胞和T细胞受体信号通路)来调节奶牛热应激的发生过程。
Abstract:
In order to explore the role of microRNA(miRNA)in the heat-stresseded response process and its regulatory mechanism when Holstein cows occurred to heating stress,the profiling of differential expressed miRNA in serum of heat-stressed and normal Holstein cows was analyzed by Solexa deep sequencing approach and real-time PCR(RT-qPCR). The target genes and signal pathway of miR-181a were predicted by bioinformatics software and database. Deep sequencing of bovine miRNA was employed to detect the differentially expressed miRNA between heat-stressed and normal Holstein cows serum. Then the expression of four selected miRNAs(miR-19a,miR-19b,miR-181a and miR-1246)in serum of heat-stressed and normal Holstein cows was also validated by RT-qPCR method. Target scan algorithm was used to predict target gene of miR-181a,and the result of gene set was analyzed by bioinformatics of GO annotations,GO term enrichment and KEGG enrichment methods. The results revealed that there were 52 miRNAs with significant differential expression between serum of heat-stressed and normal Holstein cows(P<0.01). The expression of miR-19a,miR-19b,miR-181a and miR-1246 which were also detected by RT-qPCR was in high concordance with deep sequencing results. GO function analysis showed that target genes of miR-181a were related to stress response and immune function. The results of KEGG indicated that target genes of miR-181a were mostly enriched in B cell receptor signaling pathway and T cell receptor signaling pathway. Conclusion:There are specific miRNA expression profiles in serum of heat-stressed and normal Holstein cows. Some differentially expressed miRNA in serum of heat-stressed such as miR-181a may play important roles by distinct pathway in serum of heat-stressed and normal Holstein cows.

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备注/Memo

备注/Memo:
收稿日期:2014-4-3。
基金项目:江苏省科技计划自然科学基金项目(SBK201241530);"十二五"国家科技支撑计划项目(2011BAD28B02,2012BAD12B00);中央高校基本科研业务费专项资金(KYZ201413);江苏省博士后科研资助计划项目(1302002B)
作者简介:郑月,硕士研究生。
通讯作者:李惠侠,副教授,研究方向为动物遗传育种与繁殖,E-mail:lihuixia@njau.edu.cn。
更新日期/Last Update: 1900-01-01