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当前位置: 首页 > 文章 > 海蜇(Rhopilema esculentum)磷脂酶A_2基因的cDNA、基因组克隆与表达分析 渔业科学进展 2016,37 (6) 123-130
Position: Home > Articles > cDNA, Genome Cloning, and mRNA Expression of Phospholipase A_2 Gene from the Rhopilema Esculentum Progress in Fishery Sciences 2016,37 (6) 123-130

海蜇(Rhopilema esculentum)磷脂酶A_2基因的cDNA、基因组克隆与表达分析

作  者：
杨洪;朱玲;骆晓蕊;周春娅;庄志猛
单  位：
上海海洋大学水产与生命学院;青岛海洋科学与技术国家实验室海洋生物学与生物技术功能实验室;中国水产科学研究院黄海水产研究所
关键词：
海蜇;磷脂酶A2;cDNA;基因组;表达分析
摘  要：
本研究利用RACE和RT-PCR技术克隆了海蜇(Rhopilema esculentum)磷脂酶A_2基因(Re-PLA_2-1)的cDNA及基因组序列,并分析了其m RNA在海蜇不同发育阶段的表达。Re-PLA_2-1基因的cDNA全长为824 bp,包括了48 bp的5?非编码区、504 bp的开放阅读框及272 bp的3'非翻译区。SMART分析显示,Re-PLA_2-1为分泌蛋白,包括了一个由19个氨基酸组成的信号肽和一个由118个氨基酸组成的磷脂酶A_2结构域。多序列比对和系统进化分析显示,Re-PLA_2-1基因与来自星状海葵(Nematostella vectensis)、僧袍芋螺(Conus magus)、长牡蛎(Crassostrea gigas)等磷脂酶A_2的相似性较高,共同聚类为pfam09056 GIX PLA_2分支,均包含pfam09056家族成员酶活性所必需的Ca~(2+)结合位点、活性催化位点和PLA_2结构域所必需形成二硫键的半胱氨酸。Re-PLA_2-1基因组全长为2671 bp,由4个外显子和3个内含子组成。RT-PCR结果显示,Re-PLA_2-1基因在海蜇4个发育阶段均有表达,其中,横裂体阶段的表达量最高,碟状体阶段最低。这些研究结果为进一步了解海蜇磷脂酶A_2毒素的生物功能奠定了基础。
译  名：
cDNA, Genome Cloning, and mRNA Expression of Phospholipase A_2 Gene from the Rhopilema Esculentum
作  者：
YANG Hong;ZHU Ling;LUO Xiaorui;ZHOU Chunya;ZHUANG Zhimeng;Yellow Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences;College of Fisheries and Life Science,Shanghai Ocean University;Marine Biology and Biotechnology Laboratory,Qingdao National Laboratory for Marine Science and Technology;
关键词：
Rhopilema esculentum;;Phospholipases A2;;cDNA;;Genome;;Expression analysis
摘  要：
The cDNA and gene of phospholipase A_2(Re-PLA_2-1) of Rhopilema esculentum were cloned using RACE, and the mRNA expression was monitored at different developmental stages with quantitative real-time PCR analysis. The full-length cDNA of Re-PLA_2-1 was 824 bp, containing a 5'-untranslated region(5'-UTR) of 48 bp, an open reading frame(ORF) of 504 bp, and a 3'- untranslated region(3'-UTR) of 272 bp. SMART analysis showed that Re-PLA_2-1 was a secreted protein, including a putative signal peptide consisting of 19 amino acid residues and a domain of phospholipase A_2. The deduced amino acid sequence of Re-PLA_2-1 was highly similar to those of PLA_2 s from Conus magus, Nematostella vectensis, Crassostrea gigas and so on, and they could form a cluster of pfam09056 GⅨPLA_2 revealed by the multiple sequence alignment and phylogenetic analysis. They shared the essential features of pfam09056 PLA_2 s family, including a calcium-binding site, the catalytic active sites, and a PLA_2 domain, which perfectly corresponds to the conserved disulfide-bonded cysteine residues involved in the formation of the internal disulfide. The size of Re-PLA_2-1 gene was 2671 bp that included four exons and three introns. Quantitative real-time PCR analysis revealed that the expression of Re-PLA_2-1 mRNA occurred in all four developmental stages. The expression was the highest in strobila and the lowest in ephyra. These results contributed to further understanding the biological function of PLA_2 in R. esculentum.

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