文章摘要
中华绒螯蟹内源性纤维素酶EsGH9-1基因的克隆、表达及其对不同饵料的响应*
Molecular cloning and expression analysis of endogenous cellulase EsGH9-1 from Eriocheir sinensis and its response to different diets
投稿时间:2021-08-20  修订日期:2021-10-01
DOI:
中文关键词: 中华绒螯蟹  纤维素酶  β-1,4内切葡聚糖酶  基因克隆  表达分析  不同饵料
英文关键词: Eriocheir sinensis  cellulase  β-1, 4-endoglucanase  gene cloning  expression analysis  different diet
基金项目:财政部和农业农村部:国家现代农业产业技术体系(CARS-48);“十三五”国家重点研发计划“食品安全关键技术研发”专项(2019YFC1605800);江苏省农业重大新品种创制项目(PZCZ201748);江苏省渔业科技类重点项目(D2018-4);江苏省现代农业(中华绒螯蟹)产业技术体系(JATS[2019]386);国家自然科学基金项目(31972813);江苏省研究生科研与实践创新计划项目(SJCX20_1293;SJCX20_1269)
作者单位邮编
刁澎云 江苏海洋大学海洋科学与水产学院
江苏省淡水水产研究所 
210017
储俊东 江苏海洋大学海洋科学与水产学院
江苏省淡水水产研究所 
李旭光 江苏省淡水水产研究所
江苏海洋大学海洋科学与水产学院 
陈建华 江苏省淡水水产研究所
江苏海洋大学海洋科学与水产学院
江苏省淡水水产研究所 
222005
周军 江苏省淡水水产研究所 
邓燕飞 江苏省淡水水产研究所 
许郑超 江苏省淡水水产研究所 
刘洪岩 江苏省淡水水产研究所 
曾庆飞 中国科学院南京地理与湖泊研究所 
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中文摘要:
      本研究从中华绒螯蟹转录组数据库中比对筛选到内源性纤维素酶(β-1, 4-内切葡聚糖酶)基因序列信息,通过克隆获得该纤维素酶(EsGH9-1)基因组DNA和cDNA序列。EsGH9-1基因组DNA序列长度11679 bp,包含15个外显子,14个内含子;cDNA序列全长2044 bp,编码580个氨基酸,具有糖基水解酶第9家族的典型催化功能域。RT-PCR结果显示:EsGH9-1主要分布在肝胰脏组织,在大眼幼体期和仔蟹早期高表达。不同饵料条件下,EsGH9-1相对表达量在植物性饵料组显著上调,与β-1,4内切葡聚糖酶活性趋势相一致,推测EsGH9-1参与纤维素的消化与分解。本研究首次证明中华绒螯蟹自身具有纤维素酶基因,为今后深入研究内源性纤维素酶在水生甲壳类的摄食与消化机制中的作用奠定了基础。
英文摘要:
      An endogenous cellulase (EsGH9-1) was identified from the transcriptome database of the Chinese mitten crab, Eriocheir sinensis and the genomic DNA and cDNA sequences of EsGH9-1 were cloned by PCR. The genomic DNA (11679 bp) of EsGH9-1 consists of 15 exons interrupted by 14 introns and the cDNA (2044 bp) of EsGH9-1 contains an open reading frame of 1740 bp corresponding to a polypeptide of 580 amino acid residues in which have the typical catalytic domain of glycosyl hydrolase family 9 (GHF9). Expression patterns and tissue distribution of EsGH9-1 were analyzed by RT-PCR. The results showed that EsGH9-1 was mainly distributed in hepatopancreas and was highly expressed between megalopa stage and early stage of larvae. In different diet conditions, the relative expression of EsGH9-1 was significantly up-regulated in the plant diet group, which was consistent with the trend of β-1,4-endoglucanase activity. The first finding of endogenous cellulase (EsGH9-1) gene in E. sinensis implies that EsGH9-1 may be involved in hydrolysing cellulose. Furthermore, this work is a major step toward the understanding of the role of endogenous cellulase in the digestion mechanism of aquatic crustaceans.
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