九孔鲍α-淀粉酶基因克

发布时间：2019-04-15 13:36

【摘要】：实验采用cDNA末端快速扩增(RACE)和反转录聚合酶链式反应(RT-PCR)技术对九孔鲍(Haliotis diversicolor supertexta)α-淀粉酶基因c DNA进行克隆,并分析该基因的组织表达及其与生长性状的相关性。结果表明,九孔鲍α-淀粉酶基因c DNA全长为2 260 bp,其中开放阅读框长度为2 088 bp,共编码695个氨基酸。经Singal P分析,α-淀粉酶多肽链中含有信号肽结构,且长度为18个氨基酸(MWAQYGIVSALLVLSASA),由该多肽链折叠成的三级结构中含有domain A、domain C 2个结构域,并且在domain A第28~第396氨基酸处存在催化中心。该基因在九孔鲍右侧壳肌、肠、肝脏、胃、外套膜、吻、腹足7种组织中均有不同程度表达。其中在胃中表达量最高,表达量高达326.803;而在外套膜中表达量最低,表达量仅为0.35。经ANOVA分析,消化组织与非消化组织间的α-淀粉酶基因表达量存在显著差异(P0.05)。α-淀粉酶基因mRNA表达量与生长性状呈显著正相关(P0.05)。
[Abstract]:The (Haliotis diversicolor supertexta) 伪-amylase gene cDNA of Haliotis abalone was cloned by rapid amplification of (RACE) and reverse transcription polymerase chain reaction (RT-PCR) at the end of cDNA. The tissue expression of the gene and its correlation with growth traits were analyzed. The results showed that the full length of 伪-amylase gene c DNA was 2 260 bp, and the length of open reading frame (ORF) was 2088 bp, which encodes a total of 695 amino acids. Singal P analysis showed that the 伪-amylase polypeptide chain contained signal peptide structure, and the three-order structure of 18 amino acid-length (MWAQYGIVSALLVLSASA), folded from the peptide chain contained domain A and domain C 2 domains. And there are catalytic centers at the sites of 28 ~ 396 amino acids in domain A. The gene was expressed in the right shell muscle, intestine, liver, stomach, envelope, kiss and gastropods of Haliotis abalone. The highest expression level was found in the stomach, which was up to 326.803, while the expression level in the outer membrane was the lowest, which was only 0.35%. The expression of 伪-amylase gene was significantly different between digestive tissues and non-digested tissues by ANOVA analysis (P0.05), and the expression of 伪-amylase gene mRNA was positively correlated with growth traits (P0.05).
【作者单位】：广东海洋大学水产学院;
【基金】：广东省省级科技计划项目(2015A020209196) 广东省海洋渔业科技与产业发展专项科技攻关与研发项目(A201408A04) 深圳市未来产业项目(SWCYL20150331010028)
【分类号】：S917.4

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