绵羊miR-150启动子的鉴定及生殖激素对其转录调控研究

doi:10.11843/j.issn.0366-6964.2019.07.005

Abstract

Abstract: The aim of this study was to identify the promoter of miR-150 gene and explore the effect of reproductive hormone on its transcription regulation in sheep. The miR-150 promoter region of ovary in female sheep (Small-tail Han sheep) and the reproduction-related transcription factor binding site were predicted by biological softwares, then recombinant plasmids were constructed and the dual luciferase reporter gene was used to detect the activity of miR-150 promoter. Moreover, the miR-150 promoter recombinant plasmid was transfected and miR-150 promoter activity was detected by adding FSH, LH and E₂ in serum-free medium. The miR-150 promoter region and transcription initiation site were respectively predicted using Promoter 2.0 and PROMO, and transcription factor binding sites associated with reproduction were obtained. The predicted length of the sheep miR-150 promoter fragment was 582 bp, and the miR-150 promoter plasmid was successfully constructed. The optimal transfection efficiency (liposomes:plasmid DNA=1:1) of the sheep granulosa cells and the optimal transfection proportion (50:1) of the plasmid were determined. Thus, the predicted region with significant promoter activity was verified. In addition, the activity of the miR-150 promoter was significantly down-regulated in the FSH, LH and E₂ treatment groups compared with the control group (hormone-free group) (P<0.05). The results indicated that reproductive hormone could regulate the expression of miR-150 by affecting the miR-150 promoter activity, which provided a new idea for studying the mechanism of miR-150 regulating ovine follicular development.

CLC Number:

S826.2

SONG Pengyan, LI Jiali, MIAO Yanping, LI Xiangyun, ZHOU Rongyan, XI Jianzhong. Identification of Ovine miR-150 Promoter and Its Transcriptional Regulation by Reproductive Hormone[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(7): 1358-1366.

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Identification of Ovine miR-150 Promoter and Its Transcriptional Regulation by Reproductive Hormone

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