摘要
【目的】了解大刺鳅(Mastacembelus armatus)肌肉生长抑制素(myostatin)基因mstn的序列及其在生长发育中的调控功能。【方法】利用RACE方法克隆大刺鳅mstn全长cDNA序列,采用荧光定量PCR分析其在不同组织和胚胎发育阶段的表达情况。【结果与结论】大刺鳅mstn cDNA序列全长2690 bp,包含200 bp 5′非编码区、1359 bp 3′非编码区和1131 bp开放阅读框,编码376个氨基酸。前22个氨基酸残基为信号肽,第37~254位氨基酸残基为TGF-β前肽域,第282~376位是TGF-β结构域,具蛋白酶水解位点RARR和C端生物活性区的9个保守半胱氨酸残基。系统发育分析表明,大刺鳅MSTN氨基酸序列与合鳃目、鲈形目鱼类同源性较高,与哺乳动物、鸟类、爬行类、两栖类同源性较低。实时定量PCR分析表明,大刺鳅mstn基因在各组织中均有表达,肌肉中表达量最高,眼、脑次之,鳃表达量最低;大刺鳅胚胎发育的各阶段mstn基因均有表达,囊胚期表达量最高,其次为多细胞期,出膜期表达量最低。
【Objective】To characterize the mstn cDNA sequence and study its regulatory function in growth and development of Mastacembelus armatus.【Method】Full-length mstn c DNA of of M.armatus was cloned by the RACE techniques,and quantitative real time PCR was used to analyze the expression of mstn in different embryonic development stages andt tissues.【Results】The full length c DNA mstn is 2690 bp with 200-bp 5′-untranslated region,1359-bp 3′-untranslated region,and 1131-bp open reading frame(ORF)encoding 376 amino acids pre-proprotein.The putative mstn contained a 22 amino acid signal peptide,a TGF-βpropeptide domain(37–254),a TGF-βdomain(282–376).A conserved proteolytic processing site RIRR and 9 cysteine residues in can be found in conserved locations of the protein.Phylogenetic analysis showed that the MSTN was homologous with that of the synbranchians and perciformes.However,it has low similarity to MSTN of mammals,birds,reptiles and amphibians.Quantitative real-time PCR analysis showed that the mstn gene was expressed in most tissues examined,with the highest expression detected in the muscle,followed by the eye and brain,and the lowest expression was detected in the gill.Mstn gene transcript was found in all stages of embryo development with the highest expression in blastocyst stage followed by multi-cell stage,and the lowest expression was detected in hatching stage.
作者
钟东明
赖星星
张明清
舒琥
ZHONG Dong-ming;LAI Xing-xing;ZHANG Ming-qing;SHU-Hu(School of Life Science,Guangzhou University,Guangzhou 510006,China)
出处
《广东海洋大学学报》
CAS
2020年第3期14-21,共8页
Journal of Guangdong Ocean University
基金
中国-东盟海上合作基金项目(2018.01—2020.12)
国家公益性行业(农业)重大专项(201303048)
广东省海洋渔业科技攻关与研发项目(A201601A05
2017A0007)。
关键词
大刺鳅
MSTN
基因
克隆
表达
Mastacembelus armatus
mstn
gene
cloning
expression
