生物学杂志

生物学杂志

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香鱼ApoC-Ⅰ的克隆、序列分析及其镉胁迫下的表达

  

  1. 宁波大学 海洋学院, 宁波 315211
  • 出版日期:2016-08-18 发布日期:2016-08-18
  • 通讯作者: 李长红，讲师, 主要研究方向为鱼类分子免疫学，E-mail: lichanghong0716@163.com
  • 作者简介:陈亚萍，研究方向为鱼类分子免疫学,E-mail: chen＿lh1990@163.com
  • 基金资助:
    国家自然科学基金(31372555)；高等学校博士学科点专项科研基金(20133305120008)；宁波大学学科项目(XKl15D238)

Molecular cloning, sequence analysis and expression analysis of ApoC-Ⅰgene from ayu (Plecoglossus altivelis) exposed to cadmium

  1. School of Marine Sciences, Ningbo University, Ningbo 315211, China
  • Online:2016-08-18 Published:2016-08-18

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摘要： 载脂蛋白C-Ⅰ (Apolipoprotein C, ApoC-Ⅰ)是载脂蛋白家族中分子质量最小的成员, 在脂类摄入、转运和分泌中发挥重要作用。通过cDNA文库随机测序获得了香鱼 (Plecoglossus altivelis) ApoC-Ⅰ(PaApoC-Ⅰ)全长cDNA序列。PaApoC-Ⅰ由574个核苷酸组成, 包含一个长度267 bp的开放阅读框, 编码88个氨基酸, 预测分子量为10.08 ku, N-末端1~20位氨基酸残基为信号肽序列。氨基酸序列多重比对结果表明, 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2个内含子和3个外显子组成。实时荧光定量PCR (quantitative real-time PCR, qRT-PCR)结果显示, PaApoC-Ⅰ mRNA表达量在香鱼肝组织中最高, 镉胁迫后在肝、脾、肾和肠中显著上调, 在鳃中表达量显著下调。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。

关键词: 香鱼, ApoC-Ⅰ, 序列分析, 基因表达, 镉胁迫

Abstract: ApoC-Ⅰ(Apolipoprotein C-Ⅰ) is the smallest member of the apolipoprotein family, which plays important roles in lipid uptake, secretion and transport. Here, we obtained the full-length cDNA sequence of ayu(Plecoglossus altivelis) ApoC-I gene (PaApoC-Ⅰ) through random sequencing of cDNA library. The cDNA sequence of PaApoC-Ⅰis 574 nucleotides, consisting of a 267 bp open reading frame that encodes a protein of 88 amino acids with a deduced molecular mass of 10.08 ku. 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 (qRT-PCR) analysis showed that PaApoC-ⅠmRNA had the highest expressional level in ayu liver. Upon cadmium (Cd) exposure, PaApoC-I transcripts were significantly up-regulated in ayu liver, spleen, kidney and intestine, and were significantly down-regulated in ayu gill. In summary, our present study demonstrated that a tight relationship between the PaApoC-Ⅰ expression and Cd stress, and it can be a potential biomarker for Cd exposure in aquatic organisms. The results would provide the basic data for further studying the function of fish ApoC-Ⅰ and its mechanism in fish response to heavy metal stress.

Key words: Plecoglossus altivelis, ApoC-Ⅰ, sequence analysis, gene expression, cadmium stress