摘要
为了探究肌球蛋白轻链3(Myosin light chain 3,MLC1v或MYL3)基因对骨骼肌转录调控的影响,鉴定其与肌肉生长发育的功能关系。克隆获得MYL3基因,采用生物信息学方法分析基因结构;利用荧光定量PCR技术研究MYL3基因在不同组织及不同时期的表达情况;应用免疫组化方法,定位MYL3基因在背最长肌中表达的位置;应用蛋白免疫印迹方法,研究MYL3蛋白在不同组织及不同时期的表达情况。结果表明:克隆得到布莱凯特黑牛MYL3基因(序列号:NM_001076501.2)CDS序列全长为600 bp,具有完整的阅读框,编码199个氨基酸;进化树分析发现其同源性较高;基因表达分析显示,在心脏中表达量最高,在背最长肌中表达量次之,其他组织几乎不表达,不同时期MYL3基因的表达量随月龄的增加逐渐增加;蛋白表达分析显示,MYL3蛋白主要在骨骼肌肌原纤维的粗肌丝中表达,不同时期蛋白的表达量随月龄的增加逐渐增加。因此,推测MYL3在促进骨骼肌生长发育过程中发挥重要作用。
The aim of this study was to investigate the effects of myosin light chain 3 (MLC1v or MYL3) on the regulation of skeletal muscle transcription,and to identify its functional relationship with muscle growth and development.MYL3 gene was cloned and its biological structure was analyzed by bioinformatics method.The expression of MYL3 gene in different tissues and at different stages was studied by means of fluorescence quantitative PCR technique.Immunohistochemistry showed that the expression locus of MYL3 gene were in the dorsal longissimus muscle.Western Blotting was used to study the expression of MYL3 in different tissues and at different stages.The results showed that the MYL3 gene of Black cattle (sequence number: NM_001076501.2) was cloned with a full-length reading frame of 600 bp,encoding 199 amino acids.Phylogenetic tree analysis showed that its homology was higher.Gene expression analysis showed the highest expression in the heart,followed by the longissimus muscle,and little in other tissues.The expression of MYL3 gene increased with the increase of age.The MYL3 protein mainly expressed in the myofilament of skeletal myofibrils,and increased with the age of the month at different stages.Presumably MYL3 might play an important role in skeletal muscle growth and development.
作者
刘瑞莉
吴磊
袁玮
柏学进
吕娟娟
董雅娟
LIU Ruili;WU Lei;YUAN Wei;BAI Xuejin;LU Juanjuan;DONG Yajuan(Qingdao Agricultural University,Qingdao 266109,China;Shandong Black CattleBreeding Engineering Technological Research Center,Qingdao 266109,China;Shandong Black Cattle Science Technology Co.,Ltd.,Zibo 256306,China)
出处
《华北农学报》
CSCD
北大核心
2019年第2期221-228,共8页
Acta Agriculturae Boreali-Sinica
基金
山东省现代农业产业技术体系牛产业创新团队建设项目(668/2216030)
