大珠母贝组织蛋白酶D的cDNA克隆、序列特征分析及应激表达研究  

Molecular cloning and characterization of cathepsin D in pearl oyster Pinctada maxima and its tissue expression profiles under Vibrio harveyi challenge

作　　者：潘俐玲[1,2] 黄桂菊[1] 喻达辉[1] 成书营[1,2] 王晓宁[1,2] 

机构地区：[1]农业部海水养殖生态与质量控制重点开放实验室/中国水产科学研究院南海水产研究所,广东广州510300 [2]上海海洋大学水产与生命学院,上海201306

出　　处：《广东农业科学》2011年第10期4-9,共6页Guangdong Agricultural Sciences

基　　金：国家"863"计划项目(2006AA10A415);国家现代农业产业技术体系建设专项(CARS-48);广东省科技推广项目(A200701C02;A200899C04;A200900A07);中央级公益性科研院所基本科研业务费专项(2007TS07;2009YD02)

摘　　要：通过同源克隆方法和cDNA末端快速扩增技术(RACE),获得了大珠母贝(Pinctada maxima)组织蛋白酶D基因(命名为pmCTSD)的cDNA全长序列1 742 bp,其中5’非翻译区(UTR)38 bp,3’UTR 534 bp,开放阅读框1 170 bp,编码390个氨基酸,分子量约为41.9 ku,等电点约为7.57。序列特征分析表明,pmCTSD蛋白由信号肽(Met1-Ala18)、前体域(Leu19-Lys47)和成熟域(Thr48-Asn390)3部分组成。同源性分析表明,pmCTSD氨基酸序列与其他物种高度保守,相似性介于64%～70%之间。进化分析表明,pmCTSD与栉孔扇贝亲缘关系最近,与传统分类结果基本一致。组织表达荧光定量分析发现,pmCTSD在闭壳肌、性腺、肝胰脏、外套膜和鳃组织中都有表达,但在性腺中表达量最高,肝胰脏次之。The full length of pearl oyster Pinctada maxima cathepsin D（named as pmCTSD） was determined for the first time through homologous cloning and rapid amplification of cDNA ends（RACEs） approaches.The full length cDNA contained a 5＇ UTR of 38 bp,an open reading frame（ORF） of 1 170 bp encoding 390 amino acids and a 3＇ UTR of 534 bp.The deduced amino acid residues comprised a signal peptide of 18 aa,a pro-sequence of 29 aa and a mature protein of 343 aa,with an estimated isoelectric point of 7.57 and molecular mass of 41.9 ku.Homologous analysis indicated that pmCTSD shared 64%~70% similarity in terms of amino acid sequence with other known members in CTSD subfamily.The reconstructed phylogenetic tree demonstrated that pmCTSD was closely related to that of Chlamys farreri.The tissue expression profile showed that pmCTSD expressed in all the tissues examined including adductor muscle,gonad,hepatopancreas,mantle and gill,with higher expression level in gonad and hepatopancreas.

关 键 词：大珠母贝 组织蛋白酶D 序列特征分析 哈维式弧菌刺激 组织表达 

分 类 号：S917.4[农业科学—水产科学]