华贵栉孔扇贝MEF2Cs基因克隆及表达特征分析  

Molecular cloning and expression characterization analysis of MEF2Cs in Chlamys nobilis

作　　者：朱克诚[1,2] 刘宝锁 曹明[3] 郭华阳 张楠[1] 张殿昌[1,2] ZHU Kecheng;LIU Baosuo;GUO Huayang;GUO Huayang;ZHANG Na;ZHANG Dianchang(Key Laboratory of South China Sea Fishery Resources Exploitation and Utilization,Ministry of Agriculture,South China Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences,Guangzhou 510300,China;Engineer Technology Research Center of Marine Biological Seed of Guangdong Province,Guangzhou 510300,China;The germplasm conservation center of fishery of Guangdong Province,Guangzhou 511453,China)

机构地区：[1]中国水产科学研究院南海水产研究所,农业部南海渔业资源开发利用重点实验室,广州510300 [2]广东省海洋生物种业工程技术研究中心,广州510300 [3]广东省渔业种质保护中心,广州511453

出　　处：《淡水渔业》2018年第2期32-38,共7页Freshwater Fisheries

基　　金：中国水产科学研究院南海水产研究所中央级公益性科研院所基本业务费专项资金(2014TS27)；海南省重点研发计划项目(ZDYF2016079)；国家科技基础条件平台建设运行项目(2017DKA30470)

摘　　要：肌细胞增强因子(Myocyte enhancer factor-2,MEF2)是一种肌肉特异性转录因子,对脊椎动物肌纤维的形成和发育具有重要的调控作用。为了研究MEF2对华贵栉孔扇贝(Chlamys nobilis)闭壳肌肌纤维形成和发育的调控作用,实验克隆了华贵栉孔扇贝MEF2C和MEF2C-like c DNA序列,解析了其在不同组织和发育时期的表达调控规律,分析了盐度对其表达调控的影响。结果表明,MEF2C和MEF2C-like基因开放阅读框(Open Reading Frame,ORF)长为1 302 bp和1 158 bp,分别编码433和358个氨基酸。氨基酸序列比对分析显示MADS和MEF2结构域高度同源,系统进化分析显示华贵栉孔扇贝和长牡蛎具有较近的亲缘关系。在不同组织和发育时期,华贵栉孔扇贝MEF2C和MEF2C-lik mRNA具有相似的表达规律,均在D型幼虫期及闭壳肌中显著高表达。在不同日龄的华贵栉孔扇贝闭壳肌中,MEF2Cs mRNA在60和120日龄表达水平显著升高。MEF2Cs mRNA在盐度为28时表达量最高,说明可能在此盐度下更适合华贵栉孔扇贝生长。Myocyte enhancer factor-2(MEF2)is one of the muscle specific transcription factors,which plays an important role in regulating the formation and development of vertebrate muscle fibers.cDNA sequences of MEF2C and MEF2Clike and its gene expression profile of spatio-temporal in adductor muscle were cloned and characterized in this study to investigate the regulation effect of MEF2 on the formation and development of Chlamys nobilis adductor muscle.Moreover,the expression and regulation of MEF2C and MEF2Clike by salinity was also analyzed.The results showed that MEF2C and MEF2Clike cDNA contained a 1 302 bp and 1 158 bp open reading frame(ORF)that encoded a 433 and 358 amino acids,respectively.The deduced amino acid sequences alignment results showed highly homologous between MADS and MEF2 domains.Furthermore,phylogenetic trees analysis based on amino acid datasets indicated that C.nobilis had the highest similarity with homologues of Crassostrea gigas.MEF2C and MEF2Clike showed similar expression patterns in mRNA levels,which expressed dramatically high in D-type larvae period and adductor muscle.Additionally,both of MEF2C and MEF2Clike expressed remarkable ascended in adductor muscle compared to other tissues.Expression patterns at different muscle development stages was indicated that the mRNA levels of MEF2C and MEF2Clike were observably in 60 and 120 phd(Post hatch days)in C.nobilis.Moreover,two MEF2C s transcripts at salinity of 28 were significantly higher than that at other salinity groups,which indicated that this salinity was more suitable for the growth of C.nobilis.

关 键 词：华贵栉孔扇贝(Chlamys nobilis) MEF2C和MEF2Clike 序列比对 基因表达 闭壳肌 

分 类 号：S917.4[农业科学—水产科学]