摘要
【目的】克隆陆川猪肌球蛋白调节轻链9基因(MYL9),并进行生物信息学分析及检测其在不同猪种中的表达差异,为明确陆川猪骨骼肌生长规律及研究MYL9的生理功能打下基础。【方法】根据NCBI上的家猪MYL9基因序列设计特异性引物,采用RT-PCR克隆陆川猪MYL9基因,利用在线软件对其进行生物信息学分析,并以实时荧光定量PCR检测陆川猪和杜洛克猪背最长肌中MYL9基因的表达情况。【结果】陆川猪MYL9基因编码区(CDS)序列长519bp,共编码172个氨基酸;与NCBI上已公布的家猪MYL9基因(NM_001244472.1)CDS序列比对,陆川猪MYL9基因存在2处碱基突变,分别是195 bp处C→A和498 bp处T→C,均为同义突变;陆川猪MYL9基因推导氨基酸序列与家猪MYL9基因推导氨基酸序列的同源性最高,为99.6%,与鸡的同源性最低,为88.4%。陆川猪MYL9蛋白存在3个N糖基化位点和15个磷酸化位点,不存在跨膜结构域和信号肽,符合一般EFh-PEF超家族的结构特征。陆川猪MYL9蛋白二级结构中α-螺旋占54.65%,无规则卷曲占35.47%,β-转角占8.14%,延伸链占1.74%。MYL9基因在杜洛克猪背最长肌中的相对表达量极显著高于陆川猪背最长肌(P<0.01)。【结论】MYL9基因序列在不同物种中具有较高的保守性,其在10周龄陆川猪背最长肌中的相对表达量极显著低于杜洛克猪,与其在不同猪种背最长肌中的磷酸化程度有关,也说明MYL9基因对猪肌肉生长速度有一定影响。
【Objective】In this study,myosin regulatory light chain 9 gene(MYL9)in the longissimus dorsi muscle of Luchuan pig was cloned,its bioinformatics analysis was carried out and its expressions in different pig breeds were determined in order to lay a foundation for further studying the mechanism of skeletal muscle growth of Luchuan pig and functions of MYL9.【Method】According to MYL9 gene sequence of pig on NCBI,the specific primers were designed in order to clone MYL9 gene from Luchuan pig by RT-PCR.Bioinformatics analysis on it was conducted by online software.Expression of MYL9 gene in longissimus dorsi muscle of Luchuan pig and Duroc pig was detected by fluorescence quantitative PCR.【Result】The results showed that the length of Luchuan pig MYL9 gene coding region sequence(CDS)was 519 bp and it encoded 172 amino acids.Compared with CDS sequence of pig MYL9 gene published in NCBI(NM_001244472.1),there were two synonymous mutations on 195 bp C→A and 498 bp T→C,and both were synonymous mutations.The homology between amino acid sequence of MYL9 gene in Luchuan pig and that of the published gene MYL9 in pig from NCBI were the highest,reaching 99.6%.Luchuan pig shared the lowest(88.4%)homology with amino acid sequences of Gallus.There were three N glycosylation sites,fifteen phosphorylation sites,no transmembrane domain,no signal peptide and structural features of the general EFh-PEF superfamily in MYL9 protein of Luchuan pig.The ratios of α-helix,random coil,β-turn and extending strand in the secondary structure were 54.65%,35.47%,8.14% and 1.74%,respectively.The expression of MYL9 gene in the longissimus dorsi muscle of Duroc pig was extremely higher than that of Luchuan pig(P<0.01).【Conclusion】The MYL9 gene sequence is highly conserved among different species,and its relative expression in the longissimus dorsi muscle of 10-week old Luchuan pig is extremely lower than that of Duroc pig,which is related to the degree of phosphorylation in the longissimus dorsi of different pig breeds.It also shows that the MYL9 gene has a certain effect on the growth rate of muscles in pigs.
作者
焦迪
谢婉
潘鹏丞
陈宝剑
关志惠
杨秀荣
兰干球
郭亚芬
谢炳坤
JIAO Di;XIE Wan;PAN Peng-cheng;CHEN Bao-jian;GUAN Zhi-hui;YANG Xiu-rong;LAN Gan-qiu;GUO Ya-fen;XIE Bing-kun(College of Animal Science and Technology,Guangxi University,Nanning 530004,China;Guangxi Institute of Animal Sciences/Guangxi Key Laboratory of Livestock Genetic Improvement,Nanning 530001,China)
出处
《南方农业学报》
CAS
CSCD
北大核心
2019年第3期454-460,共7页
Journal of Southern Agriculture
基金
国家自然科学基金项目(31360544)
