摘要
为了进一步研究和探索mi R-203在鸡骨骼肌发育中的作用,本研究通过对mi R-203进行靶标预测,发现真核起始因子4E(EIF4E)基因的3′UTR区存在mi R-203的潜在靶标位点。通过构建连接有EIF4E 3′UTR和靶位点突变的EIF4E 3′UTR-MUT的双荧光素酶靶标验证载体,将其分别转染到已过表达mi R-203或阴性对照鸡的DF-1细胞中,检测其中的荧光素酶活性。结果显示,mi R-203能显著抑制EIF4E 3′UTR载体的荧光素酶活性,但不影响EIF4E 3′UTR-MUT载体的荧光素酶活性,mi R-203能靶向调控鸡EIF4E的表达,研究结果为深入分析miR-203在鸡骨骼肌中的功能和作用机制奠定基础。
To further study and explore the function of miR-203 in chicken skeletal muscle development, this study predicted the potential target genes of miR-203 and found that the 3＇UTR of eukaryotic initiation factor 4E （EIF4E） gene had a potential binding site of miR-203. Next, two dual-luciferase miRNA target expression vectors which containing the EIF4E 3＇UTR or the target site mutation EIF4E 3＇lJTR-MUT were constructed, and then transfected these two vectors into chicken DF-1 cells which overexpressing miR-203 or negative control,respectively.The luciferase activity detection results showed that miR-203 could significantly inhibit the luciferase activity of the vector of EIF4E 3＇UTR,but the lueiferase activity of EIF4E 3＇UTR-MUT vector had no change. Therefore, these results demonstrated that miR-203 could regulate E1F4E expression by target binding to its 3＇UTR. This study could lay a good foundation for studying the function and mechanism of miR-203 in chicken skeletal muscle.
出处
《中国家禽》
北大核心
2015年第8期6-11,共6页
China Poultry
基金
现代农业产业技术体系建设专项资金(CARS-42-G05)
国家自然科学基金(31172200)
