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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 cDNA sequence of Amylase of Cynoglossus semilaevis with a total length of 1667 bp was obtained by RT-PCR and RACE. The open reading frame (ORF) of Amy was 1539 bp, encoding a protein of 512 amino acids. The cDNA was registered in GenBank (Accession No .: KC122678) . Homology analysis showed high homology with Tetraodon nigroviridis and Pseudopleuronectes americanus (86%, 83%). Using the neighbor-joining method to construct the phylogenetic tree, it was found that the halophila ostreatus was clustered with spotted grouper and bighead carp. The 23-day-old juvenile soft-shelled turtle fish were fed with microparticulate feed supplemented with 0%, 0.5%, 1% and 2% taurine respectively for 28 days. The relative expression of Amy gene was measured by relative quantification. The results showed that 1% group and 0.5% group were significantly higher than 0% group (P <0.05), the difference between 1% group and 0% group was extremely significant (P <0.01); 2% group and 0% group had no significant difference. The above results show that the expression of Amy gene in juvenile soft-shelled turtles is affected by the content of taurine in the feed, and the effect of 1% is significant.