小尾寒羊间情期和发情期microRNAs差异表达分析

发布时间：2018-02-01 01:46

本文关键词： 小尾寒羊间情期发情期卵巢 microRNA　出处：《畜牧兽医学报》2016年07期 　论文类型：期刊论文

【摘要】：本研究通过构建和分析小尾寒羊间情期和发情期卵巢组织microRNA(miRNA)表达谱,筛选两个时期差异表达的microRNAs,为研究microRNA调控小尾寒羊繁殖过程提供相应的理论基础。利用活体手术法在发情期(第1天)和间情期(第13天)分别采集一侧卵巢。从卵巢中提取总RNA,利用illumina Hiseq2000测序平台获取RNA数据,对表达谱和差异表达microRNAs进行生物信息学分析。实时荧光定量PCR(qRT-PCR)验证差异表达的microRNAs在小尾寒羊卵巢中表达水平。结果,成功地构建出小尾寒羊间情期和发情期microRNA的表达谱,oar-miR-99a和oar-miR-143分别是间情期和发情期表达量最高的microRNA,并筛选出在两个时期间3个显著差异表达的microRNAs,分别为oar-miR-200a、oar-miR-200b和oar-miR-200c。利用qRT-PCR对随机选择的2个显著差异表达的microRNAs进行验证,其表达水平和RNA-Seq分析结果一致。microRNA表达谱为后续的绵羊卵巢microRNA研究提供更详尽的信息。结合靶基因预测及通路富集分析,推测差异表达的microRNAs是通过代谢和免疫途径调控卵巢周期性活动。
[Abstract]:In this study, we constructed and analyzed the microRNAs miRNAs expression profiles in ovary tissues of small tail Han sheep during estrus and estrus to screen the differentially expressed microRNAs in the two stages. To provide a theoretical basis for the study of the regulation of small tail Han sheep reproductive process by microRNA. In vitro operation during estrus (1st day) and interoestrus (13th day). The total RNA was extracted from the ovary. Using illumina Hiseq2000 sequencing platform to obtain RNA data. Bioinformatics analysis of expression profile and differentially expressed microRNAs. Real-time quantitative PCR qRT-PCR. To verify the expression level of differentially expressed microRNAs in ovary of small tail Han sheep. The expression profiles of microRNA in estrus and estrus of small tail Han sheep were successfully constructed. Oar-miR-99a and oar-miR-143 were the most expressed microRNA in the period of interestrus and estrus, respectively. Three significant differences of microRNas were identified between the two periods, respectively, which were oar-miR-200a. Oar-miR-200b and oar-miR-200c. qRT-PCR were used to verify the two differentially expressed microRNAs in random selection. The microRNAs expression level is consistent with the results of RNA-Seq analysis. MicroRNAs provide more detailed information for further study of sheep ovarian microRNA. Combination of target gene prediction and pathway enrichment. Analysis. It is speculated that the differentially expressed microRNAs regulates ovarian periodic activity through metabolic and immune pathways.
【作者单位】：河北农业大学动物科技学院;
【基金】：河北省高等学校科学技术研究青年基金项目(QN2015162) 河北省首批青年拔尖人才支持计划(2013-2015) 河北农业大学青年学术带头人支持项目(2015-2017)
【分类号】：S826
【正文快照】： microRNA是一类长约22个核苷酸的非编码单链RNA分子,在哺乳动物的卵巢、输卵管、子宫、胎盘等繁殖组织器官中广泛表达[1-3]。它通过与靶mRNA特异性碱基互补配对,引起靶mRNA降解或者抑制其翻译,发挥基因转录后水平调控作用,从而参与细胞分化、增殖、凋亡等一系列生物学过程,而

【相似文献】