| 牦牛bta-miR-1434-5p和Smad1基因组织表达谱及荧光素酶报告载体构建 |
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| 引用本文： | 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.牦牛bta-miR-1434-5p和Smad1基因组织表达谱及荧光素酶报告载体构建[J].中国农业大学学报,2018,23(5):52-59. |
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| 作者姓名： | 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 |
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| 作者单位： | 西藏农牧学院动物科学学院;中国农业大学动物科学技术学院 |
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| 基金项目： | 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） |
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| 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。
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| 关 键 词： | 牦牛 bta-miR-1434-5p Smad1基因 组织表达谱 载体构建 |
| 收稿时间： | 2017/9/26 0:00:00 |
Tissue expression profiles of yak bta-miR-1434-5p and Smad1 gene and the construction of luciferase reporter vector |
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| NIU Jiaqiang,Suolangsizhu,XU Yefen,WANG Yuheng,SHANG Peng,Qiangbayangzong,GUO Ming,XI Guangying and ZHAO Liangdong.Tissue expression profiles of yak bta-miR-1434-5p and Smad1 gene and the construction of luciferase reporter vector[J].Journal of China Agricultural University,2018,23(5):52-59. |
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| Authors: | NIU Jiaqiang Suolangsizhu XU Yefen WANG Yuheng SHANG Peng Qiangbayangzong GUO Ming XI Guangying and ZHAO Liangdong |
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| Institution: | Department of Animal Science, Xizang Agriculture and Animal Husbandry College, Linzhi 860000, China,Department of Animal Science, Xizang Agriculture and Animal Husbandry College, Linzhi 860000, China,Department of Animal Science, Xizang Agriculture and Animal Husbandry College, Linzhi 860000, China,Department of Animal Science, Xizang Agriculture and Animal Husbandry College, Linzhi 860000, China,Department of Animal Science, Xizang Agriculture and Animal Husbandry College, Linzhi 860000, China,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,College of Animal Science and Technology, China Agricultural University, Beijing 100193, China and Department of Animal Science, Xizang Agriculture and Animal Husbandry College, Linzhi 860000, China |
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| Abstract: | The aim of this study was to determine the target relationship between yak Smad1 gene and bta-miR-1434-5p,establishing foundation to clarify the molecular mechanism of bta-miR-1434-5p regulation.The bta-miR-1434-5p and Smad1 mRNA expression profiles of yak tissues were detected by RT-PCR,and the targeting relationship between the 3''untranslated region (3''UTR) of Smad1 gene and bta-miR-1434-5p was explored through the construction of pmiR-RB-REPORTTM 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-1434-5p was co-expressed with Smad1 mRNA in other tissues of yak except oviduct.The 3''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-1434-5p mimics or non-target control (NC) in 293T cells,respectively,and the down-regulation effect of the luciferase expression level of the bta-miR-1434-5p mimics to wild type reporter vector of Smad1 gene was not significant.It is concluded that bta-miR-1434-5p could not target to this fragment of the 3''UTR of yak Smad1 gene. |
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| Keywords: | yak bta-miR-1434-5p Smad1 gene tissue profile vector construction |
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