半滑舌鳎(Cynoglossus semilaevis Günther)α-淀粉酶基因的克隆及牛磺酸对其表达的影响  

MOLECULAR CLONING OF α-AMYLASE GENE IN HALF SMOOTH TONGUE SOLE(CYNOGLOSSUS SEMILAEVIS GüNTHER)AND ITS EXPRESSION IN RESPONSE TO DIETARY TAURINE

作　　者：秦帮勇[1,2] 常青[2] 于朝磊[1,2] 柳淑芳[2] 马骞[2] 郑珂珂[2] 

机构地区：[1]上海海洋大学水产与生命学院,上海201306 [2]农业部海洋渔业资源可持续利用重点开放实验室中国水产科学研究院黄海水产研究所,青岛266071

出　　处：《海洋与湖沼》2013年第4期988-995,共8页Oceanologia Et Limnologia Sinica

基　　金：国家自然科学基金资助项目,31101913号

摘　　要：采用RT-PCR及RACE技术首次获得总长为1667bp的半滑舌鳎(Cynoglossus semilaevis)α-淀粉酶(Amy)基因cDNA序列,其开放阅读框为1539bp,编码512个氨基酸,已录入GenBank(登录号:KC122678)。同源性分析结果显示,与斑点绿河豚(Tetraodon nigroviridis)、美洲拟鲽(Pseudopleuronectes americanus)等同源性较高(86%、83%)。采用邻接法构建系统进化树,发现半滑舌鳎与点带石斑、鳜鱼等聚为一支。用分别添加0%、0.5%、1%、2%牛磺酸的微颗粒饲料投喂23日龄半滑舌鳎稚鱼28天,采用相对定量法检测Amy基因表达水平。结果表明,1%组和0.5%组显著高于0%组(P<0.05),1%组与0%组差异极显著(P<0.01);2%组与0%组无显著差异。上述结果表明,半滑舌鳎稚鱼Amy基因表达受饲料中牛磺酸含量影响,1%添加量作用显著。The -amylase gene （Amy gene） in Cynoglossus semilaevis was cloned in RT-PCR and RACE technique. Sequence analysis revealed a 1667bp full-length cDNA sequence containing 1539bp open reading frame （ORF） and this sequence encoded a 512 amino-acid polypeptide. Drthology analysis showed that the Amy gene of had a relatively high similarity with Tetraodon nigroviridis and Pseudopleuronectes americanus （86% and 83% respectively）. Phylogenetic analysis using neighbor-joining method on amino acid sequence showed that the Amy gene was in the same category as those in teleosteans. C. semilaevis larvae （23dph） were fed for 28 days with mircodiets which contained different taurine levels （0%, 0.5%, 1%, 2%）. Quantitative RT-PCR assay was used to estimate mRNA expression of Amy gene in response to dietary taurine. The Amy mRNA level of 1% treatment was extremely higher （P〈0.01） than that of 0% and 2% treatments, and that of 0.5% treatment was significantly higher （P〈0.05） than that of 0% and 2% treatments while there was no significant difference between 0% and 2% treatments. Therefore, these findings suggest that dietary taurine supplementation could affect the expression of Amy gene in C. semilaevis

关 键 词：半滑舌鳎 Α-淀粉酶 克隆 牛磺酸 表达 

分 类 号：Q75[生物学—分子生物学]