Identification and expression of the target gene emx2 of miR-26a and miR-26b in Paralichthys olivaceus.
Identification and expression of the target gene emx2 of miR-26a and miR-26b in Paralichthys olivaceus.
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DOI：
10.1016/j.gene.2015.06.030
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发表时间：
2015-10
期刊：
影响因子：
3.5
通讯作者：
Cui Yin;Junling Zhang;Zhiyi Shi;Wenhui Sun;Hongmei Zhang;Yuanshuai Fu
中科院分区：
文献类型：
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作者：
Cui Yin;Junling Zhang;Zhiyi Shi;Wenhui Sun;Hongmei Zhang;Yuanshuai Fu
微小RNA（microRNAs，miRNAs）通过与靶mRNA结合进一步参与多种生物学过程来调控特定基因的表达。我们前期的miRNA测序结果显示pol-miR-26 a和pol-miR-26 b在牙鲆卵巢和精巢中的表达存在性别差异。并通过生物信息学预测，将emx 2基因作为候选靶点。在本研究中，我们克隆了P. olivaceusemx 2cDNA全长741 bp，3′非翻译区912 bp，5′非翻译区12 bp。Emx 2蛋白是高度保守的，特别是其同源结构域区域从硬骨鱼到哺乳动物是100%相同的。Real-time PCR结果表明，emx 2不仅在胚胎神经胚期和成体脑组织中有高表达，而且在成体性腺中也有丰富的表达，且在卵巢中的表达量高于精巢。为了确定emx 2和miRNAs之间的关系，进行了荧光素酶报告基因分析，并验证了emx 2是pol-miR-26 a和pol-miR-26 b的共同靶基因。因此，这些数据有助于进一步阐明miR-26 a和miR-26 b参与调节性腺发育，部分是通过其目标onemx 2在P中的表达。橄榄色。
MicroRNAs (miRNAs) can regulate specific gene expression by binding to target mRNA further involution to diverse biological processes. Our previous miRNA sequencing showed that pol-miR-26a and pol-miR-26b have a sex-biased expression in ovary and testis of Paralichthys olivaceus (P. olivaceus). And the geneemptyspiracleshomeobox2(emx2) was proposed to be a candidate target by bioinformatics prediction. In this study, we cloned theP. olivaceus emx2cDNA, including a coding region of 741 bp and a 3′-untranslated region (UTR) of 912 bp and the 5′-UTR of 12 bp. The Emx2 protein is highly conserved and especially its homeodomain region is 100% identical from teleosts to mammals. Real-time PCR results showed that theemx2is not only highly expressed in embryonic neurula stage and adult brain but also has abundant expression in adult gonad, moreover, it exhibits higher expression in ovary than testis. To determine the relationship betweenemx2and miRNAs, a luciferase reporter assay was performed and verified that theemx2is a common target gene of pol-miR-26a and pol-miR-26b. These data thus helps further clarify that miR-26a and miR-26b are involved in regulating gonad development partially through its target onemx2expression inP. olivaceus.