牙鲆epigen基因的克隆以及在变态过程中的表达  

Cloning and expression of epigen gene in metamorphosing flounder Paralichthys olivaceus

作　　者：谢彩霞[1] 邢巨斌[1] 鲍宝龙[1] 

机构地区：[1]上海海洋大学省部共建水产种质资源发掘与利用教育部重点实验室,上海201306

出　　处：《上海海洋大学学报》2010年第5期577-582,共6页Journal of Shanghai Ocean University

基　　金：国家自然科学基金项目(30771668);上海市教委重点学科建设项目(J50701和S30701)

摘　　要：为了调查表皮细胞分裂原epigen基因是否与牙鲆变态发育有关,分析了从变态期牙鲆仔鱼cDNA文库中测序得到的epigen基因。发现epigen基因编码162个氨基酸,具有信号肽序列,一个跨膜结构域和EGF-like结构域。EGF-like结构域包含有可形成3个二硫键的6个半胱氨酸残基,是表皮生长因子家族的结构特征。此外,利用荧光实时定量RT-PCR,调查了epigen基因在牙鲆眼睛移动过程中的变化。相比眼睛移动之前(17DAH,day after hatching),epigen基因在眼睛移动初期(19DAH)表达明显增强,为17DAH时的2.363倍;而在眼睛移动过程的高峰期(23DAH),表达量明显回落,相对19DAH的表达量降低了80%;变态结束后(27DAH)epigen基因的表达没有重新增强或进一步的回落,提示epigen基因可能与眼睛移动初期的变态发育事件有关,而与变态高峰期和变态后期的发育事件关系不明显。To investigate the role of epigen,a member of epidermal growth factor,in flounder Paralichthys olivaceus during metamorphosis stage,we characterised the epigen gene,which is cloned from the cDNA library of metamorphosing Paralichthys olivaceus.The epigen gene encodes 162 amino acids with a signal peptide,a transmembrane helix and a EGF-like domain.EGF-like domain contains six cysteines and three disulfide bonds,and is conserved between Paralichthys olivaceus and other species.Fluorescent real-time qRT-PCR was used to analyse the change of epigen gene expression during metamorphosing stage.From pro-metamorphic stage（17DAH,day after hatching） to pre-metamorphic stage,the expression level of epigen gene increased by 136.3%.Then,the expression level decreased by around 80% from pre-metamorphic to metamorphic climax.There was no significant change from metamorphic climax to post-metamorphic stage,and the expression of epigen gene kept at a relatively low level.The expression pattern of epigen gene during metamorphosing stage indicates that the epigen might be involved in the developmental events in pre-metamorphosing flounder,but not at metamorphic climax or post-metamorphosis stage.

关 键 词：牙鲆 epigen基因 实时定量PCR 表皮生长因子 

分 类 号：S917[农业科学—水产科学]