引用本文:	周春娅,朱玲,潘滢,谢明松,杨傲傲,陈四清,庄志猛.海蜇(Rhopilema esculentum)Wnt5基因: cDNA克隆、基因组结构与表达.海洋与湖沼,,():1115-1122.

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*海蜇(Rhopilema esculentum)Wnt5基因: cDNA克隆、基因组结构与表达*
周春娅,朱玲,潘滢,谢明松,杨傲傲,陈四清,庄志猛
1.上海海洋大学水产与生命学院农业部海洋渔业可持续发展重点实验室山东省渔业资源与生态环境重点实验室中国水产科学研究院黄海水产研究所;2.农业部海洋渔业可持续发展重点实验室山东省渔业资源与生态环境重点实验室中国水产科学研究院黄海水产研究所

摘要:

采用转录组454 GS FLX测序和RACE技术, 首次解析了海蜇Wnt5基因的cDNA和基因组结构。结果表明, Re-Wnt5基因的cDNA全长1647bp, 其中编码区长1059bp, 编码了353个氨基酸的多肽; Re-Wnt5基因组含有4个外显子和3个内含子。SMART分析表明, Re-Wnt5具有Wnt家族共同的结构特征, 包括一个由20个氨基酸组成的信号肽, 2个N-糖基化位点和24个保守的参与二硫键形成的半胱氨酸。多序列比对和系统进化分析表明, Re-Wnt5基因与来自刺胞动物、无脊椎动物和脊索动物的Wnt5、脊椎动物的Wnt5a和Wnt5b具有高度相似性, Wnt5a和Wnt5b两个进化分支发生在脊索动物文昌鱼之后。实时荧光定量PCR显示, Re-Wnt5基因在海蜇四个发育阶段均有表达, 其中横裂体阶段的表达量最高, 分别是螅状体、碟状体和水母体表达量的12.38、9.99和13.01倍。关键词海蜇; Wnt5; cDNA; 基因组结构; 实时荧光定量PCR

关键词: 海蜇 Wnt5 cDNA 基因组结构实时荧光定量PCR

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Wnt5 GENE FROM RHOPILEMA ESCULENTUM: cDNA CLONING, GENOMIC ORGANIZATION AND mRNA EXPRESSION

ZHOU Chun-Ya^1,2, ZHU Ling³, PAN Ying³, XIE Ming-Song³, YANG Ao-Ao^1,2, CHEN Si-Qing³, ZHUANG Zhi-Meng³

1.College of Fisheries and Life Science, Shanghai Ocean University;2.Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Shandong Provincial Key Laboratory of Fishery Resources and Eco-environment, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences;3.Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Shandong Provincial Key Laboratory of Fishery Resources and Eco-environment, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences

Abstract:

The cDNA and genome of Wnt5 from Rhopilema esculentum were for the first time cloned by 454 GS-FLX sequencing technique and RACE technique. The results show: (1) the full-length cDNA of Re-Wnt5 was 1646bp, containing an open reading frame (ORF) of 1059bp encoding a poly-peptide with 353 amino acid residues; (2) the genome of Re-Wnt5 contained four exons and three introns. SMART analysis showed that Re-Wnt5 shared common features of Wnt family, including a putative signal peptide of 20 amino acid residues, two N-glycosylation sites and the 24 conserved cysteine residues involved in the formation of the internal disulfide bridges. The deduced amino acid sequence of Re-Wnt5 had a high homology with that of Wnt5 from Cnidaria, invertebrate and Chordata, and as well that of Wnt5 and Wnt5b from vertebrate by the multiple sequence alignment and phylogenetic analysis. The Wnt5a and Wnt5b clusters occurred after the divergence of Branchiostoma floridae. Quantitative real-time PCR analysis revealed that the expression of Re-Wnt5 transcript was detected in all four developmental stages. The mRNA expression level of Re-Wnt5 was the highest in strobila, which was 12.38-fold (P<0.01), 9.99-fold (P<0.01), and 13.01-fold (P<0.01) of that in scyphistoma, ephyra and medusa, respectively.

Key words: Rhopilema esculentum Wnt5 cDNA genomic organization real-time PCR