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以辽宁绒山羊不同发育时期皮肤毛囊组织为材料,用生物信息学方法预测miR-1298-5p的靶基因,采用RTPCR对miR-1298-5p与其潜在靶基因TGF-βR1进行核酸水平表达检测,利用Weston blot对潜在靶基因TGF-βR1进行蛋白水平表达检测。生物信息学预测结果表明TGF-βR1的3′UTR存在miR-1298-5p种子区结合位点;miR-1298-5p在皮肤毛囊不同发育时期呈现差异性表达;TGF-βR1在皮肤毛囊不同发育时期无论在核酸水平还是蛋白水平均呈现差异性表达;miR-1298-5p与其潜在靶基因之间呈现一种负调控趋势,说明miR-1298-5p可能通过负调控TGF-βR1,从而对毛囊周期性发育起到调节作用。结合靶基因预测结果、miR-1298-5p在核酸水平表达规律及TGFβR1在核酸水平表达规律和蛋白水平表达规律初步确定TGF-βR1为miR-1298-5p的靶基因。本实验旨在探讨辽宁绒山羊皮肤毛囊不同发育时期miR-1298-5p与其靶基因的调控作用及其与靶基因的潜在关系,为miR-1298-5p与其靶基因对皮肤毛囊发育作用提供理论研究数据。
The hair follicle tissue of Liaoning cashmere goats at different developmental stages was used as material to predict the target gene of miR-1298-5p by bioinformatics method. The expression of miR-1298-5p and its potential target gene TGF-βR1 was detected by RTPCR. Weston blot detection of potential target gene TGF-βR1 protein level. Bioinformatics prediction results showed that the miR-1298-5p seed-binding site exists in the 3’UTR of TGF-βR1; miR-1298-5p is differentially expressed at different developmental stages of the skin follicle; TGF- There was a negative regulation between miR-1298-5p and its potential target genes, indicating that miR-1298-5p may negatively regulate TGF-βR1 and thus inhibit hair follicle cycle Development plays a regulatory role. Combined with the predicted results of target genes, the expression regularity of miR-1298-5p at the nucleic acid level and the expression regularity of TGFβR1 at the nucleic acid level and the protein expression level initially determined that TGF-βR1 is the target gene of miR-1298-5p. The purpose of this experiment is to investigate the regulation of miR-1298-5p and its target genes in different developmental stages of Liaoning Cashmere goat skin follicles and its potential relationship with target genes to provide a theoretical study on the role of miR-1298-5p and its target genes in the development of skin follicles data.