盐藻磷脂酶C基因的克隆及生物信息学分析
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- 英文论文题名：Cloning and Bioinformatics Analysis of DsPLC Gene from Dunaliella salina
- 论文作者：柴晓杰 ; 韩冬梅 ; 丛玉婷 ; 刘艺琼 ; 岳金荣
- 英文论文作者：CHAI Xiaojie ; HAN Dongmei ; CONG Yuting ; LIU Yiqiong ; YUE Jinrong ; Key Laboratory of Hydrobiology in Liaoning Province ; Dalian Ocean University
- 年：2016
- 作者机构：大连海洋大学/辽宁省省级高校水生生物学重点实验室;
- 论文关键词：盐藻 ; 磷脂酶C ; 全长cDNA ; 生物信息学分析
- 英文论文关键词：Dunaliella salina ; Phosphatidase C ; Full length of cDNA ; Bioinformatics-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：S917.3
- 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：575k
- 原文格式：D
- 会议级别：全国
摘要
采用RT-PCR和RACE技术成功克隆了盐藻的磷脂酶C基因(GenBank Accession No.KF573428),命名为DsPLC。DsPLC基因的cDNA全长为2 628bp,其中5′UTR为48bp,3′UTR为798bp,开放阅读框为1 782bp,编码593个氨基酸。生物信息学分析表明:该蛋白为不具跨膜结构和信号肽的亲水性稳定蛋白,定位于细胞质基质;DsPLC存在两个保守区域:31-175位的氨基酸构成的X结构域和337-427位的氨基酸构成的Y结构域,其中X区46位和93位的His是其活性位点;并且存在多个潜在的磷酸化位点;二级结构的主要元件为无规卷曲和α-螺旋,催化区在三维结构上形成一个活性空腔;氨基酸序列同衣藻磷脂酶C(XP_001696502.1)具有很高的相似性(74%),进一步的进化分析结果表明,DsPLC与衣藻和团藻的磷脂酶C的亲缘关系最近。
Phospholipase C gene(GenBank Accession No. KF573428,named DsPLC)was successfully cloned by RT-PCR and RACE technology from Dunaliella salina. The full-length of cDNA sequence of DsPLC was 2 628 bp, contained a 48 bp 5′-untranslated region(UTR), a 798 bp3′-UTR, and a 1 782 bp complete ORF encoding a polypeptide of 593 amino acids.Bioinformatics analysis showed that DsPLC was a stable hydrophilic protein which had no transmembrane domains and signal peptide, located in the cytoplasmic matrix; there were two conservative regions in DsPLC: a X structure domain in 31-175 and a Y structure domain in 337-427, the His in46 and 93 of X area is its active site.And there were multiple potential phosphorylation sites; the main components of Secondary structure were random coil and alpha helix, the catalytic domain in the three-dimensional structure to form an active cavity; the amino acid sequence of DsPLC has a very high similarity(74%)with the phospholipase C of Chlamydomonas reinhardtii(XP_001696502.1), further phylogenetic analysis showed that DsPLC had the closest genetic relationship with the phospholipase C of Chlamydomonas reinhardtii and Volvox carteri.
Phospholipase C gene(GenBank Accession No. KF573428,named DsPLC)was successfully cloned by RT-PCR and RACE technology from Dunaliella salina. The full-length of cDNA sequence of DsPLC was 2 628 bp, contained a 48 bp 5′-untranslated region(UTR), a 798 bp3′-UTR, and a 1 782 bp complete ORF encoding a polypeptide of 593 amino acids.Bioinformatics analysis showed that DsPLC was a stable hydrophilic protein which had no transmembrane domains and signal peptide, located in the cytoplasmic matrix; there were two conservative regions in DsPLC: a X structure domain in 31-175 and a Y structure domain in 337-427, the His in46 and 93 of X area is its active site.And there were multiple potential phosphorylation sites; the main components of Secondary structure were random coil and alpha helix, the catalytic domain in the three-dimensional structure to form an active cavity; the amino acid sequence of DsPLC has a very high similarity(74%)with the phospholipase C of Chlamydomonas reinhardtii(XP_001696502.1), further phylogenetic analysis showed that DsPLC had the closest genetic relationship with the phospholipase C of Chlamydomonas reinhardtii and Volvox carteri.
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