栗志民, 钱佳慧, 刘建勇, 艾加林, 檀克勤. 九孔鲍α-淀粉酶基因克隆、表达分析及与生长性状相关性[J]. 南方水产科学, 2017, 13(6): 14-21. DOI: 10.3969/j.issn.2095-0780.2017.06.002
引用本文: 栗志民, 钱佳慧, 刘建勇, 艾加林, 檀克勤. 九孔鲍α-淀粉酶基因克隆、表达分析及与生长性状相关性[J]. 南方水产科学, 2017, 13(6): 14-21. DOI: 10.3969/j.issn.2095-0780.2017.06.002
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LI Zhimin, QIAN Jiahui, LIU Jianyong, AI Jialin, TAN Keqin. Molecular cloning, expression of alpha-amylase gene of Haliotis diversicolor supertexta and correlation with growth traits[J]. South China Fisheries Science, 2017, 13(6): 14-21. DOI: 10.3969/j.issn.2095-0780.2017.06.002
Citation: LI Zhimin, QIAN Jiahui, LIU Jianyong, AI Jialin, TAN Keqin. Molecular cloning, expression of alpha-amylase gene of Haliotis diversicolor supertexta and correlation with growth traits[J]. South China Fisheries Science, 2017, 13(6): 14-21. DOI: 10.3969/j.issn.2095-0780.2017.06.002
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九孔鲍α-淀粉酶基因克隆、表达分析及与生长性状相关性

Molecular cloning, expression of alpha-amylase gene of Haliotis diversicolor supertexta and correlation with growth traits

    摘要
  • 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cDNA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cDNA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SingalP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（P < 0.05）。α-淀粉酶基因mRNA表达量与生长性状呈显著正相关（P < 0.05）。

     

    Abstract: We cloned the alpha-amylase gene cDNA of Haliotis diversicolor supertexta by rapid amplification of cDNA ends method (RACE) and reverse transcription polymerase chain reaction (RT-PCR) to analyze its tissue expression and correlation with growth traits. The results show that the full length of alpha-amylase gene cDNA was 2 260 bp with an open reading frame (ORF) of 2 088 bp encoding 695 amino acids. The alpha-amylase contained a signal peptide with 18 amino acid residues (MWAQYGIVSALLVLSASA). Two functional structure domains (domain A, domain C) were found in the protein of alpha-amylase. Domain A contained one catalytic center between 28th and 396th amino acids. The alpha-amylase mRNA expression in seven tissues (adductor muscle, intestine, digestive gland, stomach, mantle, rostral, foot) was observed, highest in stomach (326.803) and lowest in mantle (0.35). Significant difference was found in the expression of alpha-amylase between digestive and non-digestible tissues (P < 0.05). Significant positive correlation existed between alpha-amylase gene expression and growth traits (P < 0.05).

     

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