ssc-miR-196a靶向PPARGC1A基因影响猪骨骼肌发育
ssc-miR-196a Affects Porcine Skeletal Muscle Development by Regulating PPARGC1A Gene Expression
张展境 1李淑怡 1张瑞 1郑美丽 2林涛 1董新星 3韩海银 1白莹1
作者信息
- 1. 河北工程大学生命科学与食品工程学院,河北 邯郸 056038
- 2. 北京市畜牧总站,北京 100107
- 3. 云南农业大学动物科学技术学院,云南 昆明 650201
- 折叠
摘要
该研究运用miRsystem和MiRWalk 2个在线软件筛选与PPARGC1A基因具有潜在调控关系的microRNA;构建psiCHECK2-PPARGC1A-3'UTR-WT和psiCHECK2-PPARGC1A-3'UTR-MUT双荧光素酶报告载体,并检测候选microRNA与PPARGC1A基因间的靶向调控关系.结果显示,ssc-miR-196a与PPARGC1A基因具有潜在的调控关系,ssc-miR-196a可与PPARGC1A 3'UTR区结合并显著降低细胞中荧光素酶的活性(P＜0.01),ssc-miR-196a在金华猪90日龄和180日龄背最长肌中的表达水平略高于大白猪,ssc-miR-196a在180日龄猪背最长肌中表达水平显著高于30日龄和90日龄(P＜0.05).ssc-miR-196a靶向结合PPARGC1A基因3'UTR,为进一步开展microRNA介导PPARGC1A基因调控猪骨骼肌发育方面的研究提供参考.
Abstract
In this study,two online programs,miRsystem and MiRWalk,were used to screen microRNAs that have potential regulatory relationships with the PPARGC1A gene.The reporter vectors psiCHECK2-PPARGC1A-3'UTR-WTand psiCHECK2-PPARGC1A-3'UTR-MUT were constructed,and the dual-luciferase assay was used to detect the target-regulatory relationship between the candidate microRNAs and PPARGC1A gene.The results revealed a potential regulatory relationship between ssc-miR-196a and the PPARGC1A gene.Dual luciferase assay showed that miR-196a could bind to the 3'UTR of PPARGC1A and significantly reduced luciferase activity in cells(P＜0.01).The expression level of ssc-miR-196a in longissimus muscle of Jinhua pigs at age 90 and 180 days was slightly higher than that of Large White pigs,and its expression level in longissimus muscle of pigs at age 180 days was significantly higher than that of pigs at age 30 and 90 days(P＜0.05).The specific binding of ssc-miR-196a to the 3'UTR of PPARGC1A,which provides a reference for further research into the regulation of porcine skeletal muscle development by microRNA-mediated PPARGC1A gene.
关键词
ssc-miR-196a/PPARGC1A/背最长肌/猪Key words
ssc-miR-196a/PPARGC1A/longissimus dorsi muscle/porcine引用本文复制引用
基金项目
河北省自然科学基金(C2021402038)
国家科技重大专项(2018ZX0800928B)
云南省农业联合专项(202101BD070001-103)
出版年
2024
国家科技期刊平台
NSTL
万方数据