摘要
本研究旨在探究甲基转移酶METTL21C在家禽骨骼肌发育分化过程中的作用。采用实时荧光定量PCR (quantitative Real-time PCR, qPCR)检测METTL21C基因在鸡不同组织中的表达情况,绘制其组织表达谱;选取3个时间点,检测其在骨骼肌组织中的表达情况。通过酶消化法从鸡骨骼肌组织中分离得到原代细胞;将METTL21C超表达载体转染至原代鸡骨骼肌细胞,通过qPCR和Western blotting检测Pax7、MyoD、Myf5、MyoG等基因的表达水平。结果显示,METTL21C在心肌和骨骼肌组织中的表达量显著高于其他组织,在胚胎期和幼龄期骨骼肌中的表达呈上升趋势;超表达METTL21C后,成肌相关基因Pax7、MyoD、Myf5、MyoG的表达量显著升高。本研究初步发现甲基转移酶METTL21C具有促进家禽骨骼肌发育分化的作用,为骨骼肌发育的分子机理的研究及相关医学研究提供数据支持。
The aim of this study was to inves tigate the role of METTL21 C gene in the development and differentiation of skeletal muscle in poultry.The relative mRNA expression of METTL21 C in different tissues of chicken was detected by real-time fluorescence quantitative PCR to generate the expression profile.The expression level of METTL21 C gene was tested in skeletal muscle tissue at three timing.Primary cells were isolated from chicken skeletal muscle tissue by enzyme digestion.Overexpression vector of METTL21 C gene was transfected into primary skeletal muscle cells.The expression of Pax7,MyoD,Myf5 and MyoG genes were detected by q PCR and Western blotting.Expression levels of METTL21 C gene in cardiac muscle and skeletal muscle were much higher than that of other tissues,and presented an upward trend in skeletal muscle embryonic stage and juvenile stage.The expression of muscle-related genes Pax7,MyoD,Myf5 and MyoG was increased significantly with overexpression of METTL21 C.This study found that METTL21 C can promote the development and differentiation of skeletal muscle in poultry,which could provide data support for the molecular mechanism of skeletal muscle development and corresponding medical research.
作者
李蕊清
尹美辰
赵家荣
王令
张涛
路宏朝
Li Ruiqing;Yin Meichen;Zhao Jiarong;Wang Ling;Zhang Tao;Lu Hongzhao(School of Biological Science Technology and Engineering,Shaanxi University of Technology,Hanzhong,723001)
出处
《基因组学与应用生物学》
CAS
CSCD
北大核心
2019年第6期2503-2508,共6页
Genomics and Applied Biology
基金
陕西省农业科技创新与攻关项目(2016NY-084)
陕西理工大学人才启动基金项目(SLGQD2017-09)共同资助
