绒山羊皮肤毛囊miR-1298-5P靶基因预测及表达载体构建

发布时间：2018-02-28 22:21

本文关键词： 辽宁绒山羊皮肤毛囊 miR--P FgF 靶基因预测表达载体构建　出处：《中国畜牧杂志》2017年01期 　论文类型：期刊论文

【摘要】：探讨辽宁绒山羊皮肤毛囊不同发育时期miR-1298-5P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依据。以辽宁绒山羊皮肤毛囊组织为材料,通过miRNA的分离、表达、靶基因预测筛选表达及靶位点3'UTR表达载体构建,采用RT-PCR进行表达检测,用生物信息学方法预测miR-1298-5P的靶基因,并对其靶基因Fg F2的靶位点进行克隆测序、缺失突变。miR-1298-5P及其靶基因在皮肤毛囊不同发育时期均呈现差异性表达,二者之间呈现一种负调控趋势,其可能对皮肤毛囊周期性发育起到一定的调控作用。靶基因预测发现Fg F2的3'UTR存在mi R-1298-5P种子区结合位点,并成功构建了Fg F2 3'UTR表达载体,该载体的成功构建为后期过表达转染体系的建立和功能基因的验证奠定了基础。
[Abstract]:To investigate the regulation of miR-1298-5P and its potential relationship with target gene in Liaoning cashmere goat skin follicle at different developmental stages, and to provide a theoretical basis for the effect of miR-1298-5P and its target gene on the development of skin hair follicle. The skin hair follicle tissue of Liaoning cashmere goat was used as the material. Through the isolation, expression, target gene prediction, screening and expression of miRNA and the construction of target 3UTR expression vector, the target gene of miR-1298-5P was detected by RT-PCR, the target gene of miR-1298-5P was predicted by bioinformatics, and the target of FgF2 was cloned and sequenced. The deletion mutation. MiR-1298-5P and its target gene showed different expression at different stages of skin hair follicle development, and there was a trend of negative regulation between them. It may play a role in regulating the periodic development of skin hair follicles. The target gene predicted that the 3UTR of FgF2 had a seed region binding site of mi R-1298-5P, and successfully constructed the expression vector FgF23 UTR. The successful construction of the vector laid a foundation for the establishment of the late overexpression transfection system and the verification of functional genes.
【作者单位】：吉林农业大学动物科学技术学院;长春科技学院;
【基金】：吉林省科技发展计划项目(20150236) 吉林省教育厅项目(2015566)
【分类号】：S827

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