香鱼ApoC-Ⅰ基因的克隆、序列分析及其镉胁迫下的表达
Molecular Cloning, Sequence Analysis and Expression Analysis of ApoC-Ⅰgene from ayu (Plecoglossus altivelis) Exposed to Cadmium
载脂蛋白C-Ⅰ (Apolipoprotein C, ApoC-Ⅰ)是载脂蛋白家族中分子量最小的成员, 在脂类摄入、分泌和转运中发挥重要作用. 本研究从香鱼 (Plecoglossus altivelis)肝组织cDNA文库中获得了香鱼ApoC-Ⅰ基因 (PaApoC-Ⅰ)全长cDNA序列. PaApoC-Ⅰ由574个核苷酸组成, 包含一个大的开放阅读框, 编码88个氨基酸, 预测分子量为10.08 kDa, N-末端含一段由20个氨基酸残基的信号肽序列. 氨基酸序列多重比对分析表明, PaApoC-Ⅰ与虹鳟 (Oncorhynchus mykiss) ApoC-Ⅰ同源性最高, 为67.0%; 系统进化树分析表明, 哺乳动物、爬行动物、两栖类和鱼类的 ApoC-Ⅰ分别成簇, PaApoC-Ⅰ位于鱼类 ApoC-Ⅰ簇中, 且与虹鳟、ApoC-Ⅰ进化相关性最高. 基因组结构分析表明,PaApoC-Ⅰ基因组DNA序列由3个外显子和2个内含子组成. 实时荧光定量PCR (quantitative real-time PCR, qRT-PCR)结果显示, PaApoC-Ⅰ基因mRNA在肝组织中表达量最高; 镉胁迫后香鱼肝、脾、肾和肠中PaApoC-ⅠmRNA表达量显著上调, 鳃组织中PaApoC-Ⅰ基因mRNA表达量显著显著下调. 综上, PaApoC-Ⅰ的表达与镉胁迫密切相关, 可能作为水环境中重金属镉污染的生物分子标记. 研究结果为进一步研究鱼类ApoC-Ⅰ的功能及其在重金属胁迫中的作用机制提供了基础资料.
poC-Ⅰ(Apolipoprotein C, ApoC-Ⅰ) is the smallest of the apolipoprotein family, which plays important roles in lipid uptake, secretion and transport. Here, we obtained full-length cDNA sequence of ayu ApoC- I gene (PaApoC-Ⅰ) from ayu (Plecoglossus altivelis) liver tissue cDNA library. The cDNA sequence of PaApoC-Ⅰis 574 nucleotides, containing a large open reading frame that encodes a protein of 88 amino acids with a deduced molecular mass of 10.08 kDa. The N-terminus of PaApoC-Ⅰcontains a predicted signal peptide of 20 amino acids. Multiple sequence alignment of complete amino acid sequences showed that PaApoC-Ⅰshared the highest amino acid sequence identity (67.0%) with that of rainbow trout (Oncorhynchus mykiss). Phylogenetic tree analysis revealed that ApoC-Ⅰfrom mammals, reptiles, amphibians and fish formed different clusters, and PaApoC-Ⅰfalled into the fish ApoC-Ⅰcluster and was most closely related to that of rainbow trout. Genome structure analysis showed that PaApoC-Ⅰcontained three exons and two introns. Quantitative real-time PCR (quantitative real-time PCR, qRT-PCR) analysis showed that PaApoC-ⅠmRNA had the highest expressional level in ayu liver. Upon Cd exposure, PaApoC- I transcripts siginificantly up-regulated in liver, spleen, kidney and intestine, and significantly down-regulated in gill. In summary, our present study demonstrated that a tight relationship between the PaApoC-Ⅰexpression and cadmium stress, and it can be a potential biomarker of Cd exposure in aquatic organisms. The results provided the basic data for further studying the function of fish ApoC-Ⅰand its mechanism in fish response to heavy metal stress.
陈亚萍、丁斐斐、陈炯、李长红
分子生物学环境生物学生物科学现状、生物科学发展
水产香鱼ApoC-Ⅰ序列分析基因表达镉胁迫
aquaculturePlecoglossus altivelisApoC-ⅠSequence analysisGene expressionCadmium stress
陈亚萍,丁斐斐,陈炯,李长红.香鱼ApoC-Ⅰ基因的克隆、序列分析及其镉胁迫下的表达[EB/OL].(2015-10-15)[2025-08-02].http://www.paper.edu.cn/releasepaper/content/201510-81.点此复制
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