应用Solexa测序技术挖掘山羊卵巢组织microRNA

MicroRNAs in Ovaries of Goats (Capra hircus) Identified by Solexa Sequencing

【目的】旨在构建、分析山羊卵巢组织miRNA表达谱,为进一步研究特定miRNA与山羊卵泡发育及性激素分泌等相关生殖活动的关系奠定理论基础。【方法】首先从山羊卵巢组织总RNA中分离小片段RNA,然后进行Solexa测序和生物信息学分析,最后利用q-PCR技术验证miRNA在山羊卵巢组织中的表达。【结果】共鉴定出508个山羊卵巢组织表达的miRNA和19个对应的miRNA*,这些miRNA在哺乳动物(绵羊、牛、猪、马、狗)进化过程中高度保守;q-PCR验证了8个miRNA在山羊卵巢组织中的表情况,定量结果与测序结果一致。【结论】成功构建了山羊卵巢组织miRNA表达文库,山羊卵巢组织miRNA表达丰富且其表达量各异。

[Objective] Characterization ofmicroRNA expression in hircine ovary tissues was studied, with the purposes of providing basic information for further studies of relationships between specific miRNAs and hircine reproduction such as folliculogenesis and sex hormone secretion. [ Method ] The small RNAs isolated from total RNA of ovary tissues were sequenced by Solexa and then bioinformatics analysis were perfomed. Meanwhile, the expression of selected miRNAs was validated by q-PCR. [Result] Sequence analysis indicated that 508 ovary miRNAs and 19 corresponding miRNA＊s were identified which are conservative in evolution of mammals （sheep, bovine, swine, horse, dog）. The expression of 8 selected miRNAs in ovary tissues obtained by q-PCR was in agreement with Solexa sequencing results. [ Conclusion ] The expression profiling of miRNAs, which are abundant and differentially expressed in hircine ovary tissues, was constructed successfully.