坛紫菜过氧化氢酶基因的克隆及表达特征
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集美大学水产学院,集美大学水产学院,集美大学水产学院,集美大学水产学院,集美大学水产学院,集美大学水产学院,集美大学水产学院

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国家“八六三”高技术研究发展计划(2012AA10A411);国家自然科学基金(41176151,41276177);福建省自然科学基金(2014J05041,2014J07006)


Cloning and expression pattern analysis of catalase (CAT) gene from Pyropia haitanensis
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College of Fisheries,Jimei University,College of Fisheries,Jimei University,College of Fisheries,Jimei University,College of Fisheries,Jimei University,College of Fisheries,Jimei University,College of Fisheries,Jimei University,College of Fisheries,Jimei University

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    摘要:

    过氧化氢酶(CAT)是植物细胞抗氧化酶的主要成员,在逆境胁迫下的活性氧(ROS)清除中发挥着重要的作用。本研究采用RACE扩增技术克隆获得了坛紫菜CAT基因的全长,被命名为PhCAT(Genbank收录号:KM655303)。该基因全长1983 bp,可编码包含542个氨基酸的多肽,与条斑紫菜CAT基因的序列相似性为83.08%。多序列比对和系统进化树分析结果确认PhCAT属于CAT基因家族。PhCAT基因定量分析结果显示,失水胁迫条件下,PhCAT的基因表达水平只在失水率≥60%时才显著上调;不同时间水平的高温胁迫条件均可诱发PhCAT基因的上调表达。而CAT酶活测定结果却显示,各种水平的失水胁迫条件下,坛紫菜细胞内的CAT酶活水平均显著低于正常条件下的酶活水平;高温胁迫24 h时,CAT酶活水平显著增强,而其余时间点的酶活水平则显著降低。研究表明CAT酶在坛紫菜高温胁迫应答中发挥着重要作用,但在失水胁迫应答中却没有发挥明显作用。

    Abstract:

    Catalase (CAT) is a main member of the plant cell anti-oxidant enzymes, which plays an important role in clearing reactive oxygen species (ROS) under the stressful conditions. In this study, the full-length sequence of CAT gene of Pyropia haitanensis was obtained by rapid amplification of cDNA ends (RACE), which was named PhCAT (GenBank accession: KM655303). Full-length of the gene is 1983 bp which could code a 542 amino acid polypeptide. The sequence similarity was 83.08% when compared with the CAT gene from P. yezoensis. Multiple sequence alignment and phylogenetic tree analysis results confirmed that PhCAT belongs to the CAT gene family. PhCAT gene quantitative analysis found the gene expression levels rose significantly when water loss equal or greater than 60%. The CAT expressions rose under high temperature stress (29℃) regardless of the treatment time. However, the responses of CAT activity showed different trends: CAT activities were significantly decreased by all levels of water loss; but only significantly enhanced under 24 h of high temperature treatment, while at the rest of the time point, enzyme activity level decreased significantly. All of these results suggested that the CAT enzyme plays an important role in responses of P. haitanensis to high temperature stress, but did not exhibit significant effects under the water loss stress.

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仵燕青,肖海东,许凯,徐燕,纪德华,陈昌生,谢潮添.坛紫菜过氧化氢酶基因的克隆及表达特征[J].水产学报,2016,40(10):1576~1585

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  • 收稿日期:2015-10-25
  • 最后修改日期:2016-05-28
  • 录用日期:2016-10-20
  • 在线发布日期: 2016-10-21
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