厚壳贻贝细胞外调节蛋白激酶基因McERK的克隆及其在附着变态中的作用
Cloning of extracellular regulated protein kinases gene McERK and its role in larval settlement and metamorphosis in mussel Mytilus coruscus
刘甜甜 1侯金杞 1马蕃 1杨金龙 1梁箫1
作者信息
- 1. 上海海洋大学水产与生命学院,国家海洋生物科学国际联合研究中心,上海 201306;上海市水产动物良种创制与绿色养殖协同创新中心,上海 201306;中国-葡萄牙星海"一带一路"联合实验室,上海 200120
- 折叠
摘要
为解析细胞外调节蛋白激酶(extracellular regulated protein kinases,ERK)基因(McERK)在厚壳贻贝(Mytilus coruscus)附着变态过程中的作用机制,采用RACE技术、系统进化分析和Real-time PCR等方法,对McERK基因分子特点和表达情况进行了研究.结果表明:McERK基因序列全长为1 236 bp,开放阅读框为846 bp,可编码281个氨基酸,具有典型的丝氨酸/苏氨酸蛋白激酶结构域S_TKc;系统进化树分析显示,厚壳贻贝与加州贻贝(M.californianus)、地中海贻贝(M.galloprovincialis)和欧洲贻贝(M.edulis)的ERK聚为一支,且与欧洲贻贝序列的一致性最高(74.27％);荧光定量PCR分析显示,McERK基因在厚壳贻贝成贝各组织中广泛表达,在外套膜、唇瓣、鳃和血细胞中均有较高的表达量,其在担轮幼虫到稚贝的各个发育阶段均有表达,且在眼点幼虫阶段的表达量显著高于稚贝阶段(P＜0.05);利用RNA干扰技术敲降眼点幼虫McERK基因后,幼虫变态率显著降低(P＜0.05).研究表明,McERK基因可能参与调控厚壳贻贝幼虫附着变态过程,本研究结果可为探究McERK调控厚壳贻贝的发育过程提供数据支持.
Abstract
In order to analyze the mechanism of extracellular regulated protein kinases gene McERK in the settle-ment and metamorphosis of mussel Mytilus coruscus,the molecular characteristics and spatial and temporal expres-sion of McERK gene were analyzed by RACE technology,phylogenetic analysis and Real-time PCR.The results showed that the full length of McERK gene sequence was 1 236 bp,the open reading frame(ORF)was 846 bp,encoding 281 amino acids,with a typical serine/threonine protein kinase domain S_TKc.Phylogenetic tree analysis revealed that the gene was clustered with mussels M.californianus,M.galloprovincialis and M.edulis,with the maxi-mal similarity with M.edulis(74.27％).The analysis of real-time quantitative PCR temporal and spatial expression of McERK gene by real-time quantitative PCR showed that McERK gene was widely expressed in mantle,lip,gill and blood cells of adult mussel from trochophore larvae to juveniles,significantly higher in the pediveliger larvae stage than that in the juvenile stage(P＜0.05).The metamorphosis rate of larvae was significantly reduced in pedi-veliger larvae exposed to knocking down the McERK gene by RNA interference technology,indicating that McERK gene participated in the regulation of the settlement and metamorphosis of M.coruscus larvae.The finding provides data support for exploring of the regulation of McERK gene in the development process of M.coruscus.
关键词
厚壳贻贝/ERK基因/幼虫变态/基因克隆Key words
Mytilus coruscus/ERK/larval metamorphosis/gene cloning引用本文复制引用
基金项目
国家重点研发计划(2022YFE0204600)
国家重点研发计划(2023YFE0115500)
出版年
2024
NETL
NSTL
万方数据