摘要
为探索牦牛Smad1基因与bta-miR-1434-5p是否存在靶向关系,探究bta-miR-1434-5p分子调控可能机制,利用RT-PCR技术对bta-miR-1434-5p和Smad1 mRNA在牦牛组织中的表达谱进行检测,并构建pmiR-RBREPORTTM双荧光素酶报告载体对牦牛Smad1基因3′端非翻译区(3′UTR)与bta-miR-1434-5p的靶向关系进行研究。结果表明:Smad1基因mRNA在牦牛下丘脑、垂体、心、肝、脾、肺、肾、骨骼肌、淋巴结、卵巢、输卵管、子宫组织中均有广泛表达;bta-miR-1434-5p除在牦牛输卵管中没有表达外,在其他组织均与Smad1基因mRNA共表达;获得牦牛Smad1基因3′UTR序列并成功构建野生型及突变型pmiR-RB-REPORTTM双荧光素酶报告质粒;荧光素酶活性检测发现bta-miR-1434-5p mimics对Smad1野生型质粒报告荧光没有明显的下调作用,由此推测btamiR-1434-5p与牦牛Smad1基因的该段3′UTR之间未发现明显互作。
The aim of this study was to determine the target relationship between yak Smad1 gene and bta-miR-1434-5 p,establishing foundation to clarify the molecular mechanism of bta-miR-1434-5 p regulation.The bta-miR-1434-5 p and Smad1 mRNA expression profiles of yak tissues were detected by RT-PCR,and the targeting relationship between the3′untranslated region（3′UTR）of Smad1 gene and bta-miR-1434-5 p was explored through the construction of pmiR-RBREPORTTM dual-luciferase reporting vector.The results showed that：Smad1 mRNA was widely expressed in many tissues of yak such as hypothalamus,pituitary,heart,liver,spleen,lung,kidney,skeletal muscle,lymph node,ovary,oviduct and uterus;bta-miR-1434-5 p was co-expressed with Smad1 mRNA in other tissues of yak except oviduct.The3′UTR of yak Smad1 gene sequence was obtained and the wild and mutant type of pmiR-RB-REPORTTM dual luciferase reporting vectors were successfully constructed.Luciferase activity was detected after co-transfection of the wild or mutant type vector with bta-miR-1434-5 p mimics or non-target control（NC）in 293 Tcells,respectively,and the downregulation effect of the luciferase expression level of the bta-miR-1434-5 p mimics to wild type reporter vector of Smad1 gene was not significant.It is concluded that bta-miR-1434-5 p could not target to this fragment of the 3′UTR of yak Smad1 gene.
作者
牛家强
索朗斯珠
徐业芬
王玉恒
商鹏
强巴央宗
郭敏
席广银
赵良栋
NIU Jiaqiang;Suolangsizhu;XU Yefen;WANG Yuheng;SHANG Peng;Qiangbayangzong;GUO Ming;XI Guangying;ZHAO Liangdong(Department of Animal Science, Xizang Agriculture and Animal Husbandry College, Linzhi 860000, China;College of Animal Science and Technology, China Agricultural University, Beijing 100193, China)
出处
《中国农业大学学报》
CAS
CSCD
北大核心
2018年第5期52-59,共8页
Journal of China Agricultural University
基金
国家自然科学基金项目(31460604)
中西部高校综合实力提升计划:西藏农牧学院动物遗传育种与繁育学科建设(502000105)
国家肉牛牦牛产业体系项目(CARS-37)
