中华绒螯蟹纤维素酶基因的克隆与表达分析
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- 英文论文题名：Cloning and expression profiling of a cellulose gene in the Chinese mitten crab(Eriocheir sinensis)
- 论文作者：李旭光 ; 周刚 ; 周军 ; 林海 ; 李跃华 ; 陆全平 ; 付龙龙
- 英文论文作者：LI Xuguang ; ZHOU Gang ; ZHOU Jun ; LIN Hai ; LI Yuehua ; LU Quanping ; Fu Longlong ; Freshwater Fisheries Ressearch Institute of Jiangsu Province
- 年：2016
- 作者机构：江苏省淡水水产研究所;
- 论文关键词：中华绒螯蟹 ; 纤维素酶基因 ; 内切β-1 ; 4葡聚糖酶 ; 克隆 ; 表达分析
- 英文论文关键词：Eriocheir sinensis ; cellulose ; endo-1 ; 4-glucanase ; cloning ; expression analysis
- 会议召开时间：2016-11-02
- 会议录名称：2016年中国水产学会学术年会论文摘要集
- 语种：中文
- 分类号：S917.4
- 学会代码：OGSB
- 会议名称：2016年中国水产学会学术年会
- 会议地点：中国四川成都
- 主办单位：中国水产学会、四川省水产学会
- 学会名称：中国水产学会
- 页数：1
- 文件大小：574k
- 原文格式：D
- 会议级别：全国
摘要
本实验采用cDNA末端快速扩增技术(RACE)从中华绒螯蟹体内克隆了一个纤维素酶(内切β-1,4葡聚糖酶)基因cDNA序列全长,并通过RT-PCR技术研究了该基因在中华绒螯蟹不同组织与不同发育阶段的表达特征。结果表明中华绒螯蟹纤维素酶基因cDNA全长1971bp,开放阅读框架1722bp,编码573个氨基酸。该纤维素酶基因cDNA具有与已发现的纤维素酶基因相同的保守区结构和功能域,并且与其它已知甲壳类的纤维素酶有较高的同源性,含有糖基水解酶第9家族的保守结构域。RT-PCR结果显示该纤维素酶基因主要在肝胰脏、肌肉和表皮组织中表达,其中在肝胰脏组织中表达量最高。对不同发育阶段的该纤维素酶基因表达分析发现,该基因从蚤状幼体到成蟹阶段均有表达,推测其在中华绒螯蟹生长过程中对纤维素酶类食物的消化与降解起到重要作用。
In the present study, a cellulose(endo-1,4-glucanase) gene was cloned from the Chinese mitten crab(Eriocheir sinensis) by the rapid amplification of cDNA end(RACE) methods. The expression of the gene in different tissues and different developmental stages of E. sinensis was analyzed by real-time quantitative PCR. The full-length of endo-1,4-glucanase cDNA is 1971 bp,containing an open reading frame(ORF) of 1722 bp which encodes a peptide of 573 amino acids.Phylogenetic tree revealed that the endo-1,4-glucanase was closely related to known endo-1,4-glucanase of other crustaceans, and it has a glycosyl hydrolase family 9(GHF9)-like conserved domain with all catalytically important residues conserved. The result showed that the endo-1,4-glucanase gene was mainly expressed in the hepatopancreas, with low levels of expression in the muscle and cuticle. The endo-1,4-glucanase gene was expressed in different developmental stages including zoea larvae, megalopa larvae,juvenile crab and adult crab which indicated that the gene may play a role in the digestion and degradation of cellulose food during the growth process of E. sinensis..
In the present study, a cellulose(endo-1,4-glucanase) gene was cloned from the Chinese mitten crab(Eriocheir sinensis) by the rapid amplification of cDNA end(RACE) methods. The expression of the gene in different tissues and different developmental stages of E. sinensis was analyzed by real-time quantitative PCR. The full-length of endo-1,4-glucanase cDNA is 1971 bp,containing an open reading frame(ORF) of 1722 bp which encodes a peptide of 573 amino acids.Phylogenetic tree revealed that the endo-1,4-glucanase was closely related to known endo-1,4-glucanase of other crustaceans, and it has a glycosyl hydrolase family 9(GHF9)-like conserved domain with all catalytically important residues conserved. The result showed that the endo-1,4-glucanase gene was mainly expressed in the hepatopancreas, with low levels of expression in the muscle and cuticle. The endo-1,4-glucanase gene was expressed in different developmental stages including zoea larvae, megalopa larvae,juvenile crab and adult crab which indicated that the gene may play a role in the digestion and degradation of cellulose food during the growth process of E. sinensis..
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