生物学杂志

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香鱼HSP90β的克隆、序列特征及表达分析

  

  1. 宁波大学 海洋学院, 宁波 315211
  • 出版日期:2018-02-18 发布日期:2018-02-18
  • 通讯作者: 李长红, 博士, 硕士生导师, 研究方向为水产免疫学,E-mail: lichanghong0716@163.com
  • 作者简介:默立宾, 主要研究方向为水产免疫学,E-mail: molibin777@163.com
  • 基金资助:
    国家自然科学基金(31372555, 31402333); 浙江省自然科学基金一般项目(Y17C190003); 宁波大学校学科项目(XKl15D238)

Molecular cloning, sequence characterization and expression analysis of HSP90β from ayu (Plecoglossus altivelis)

  1. School of Marine Sciences, Ningbo University, Ningbo 315211, China
  • Online:2018-02-18 Published:2018-02-18

摘要: 热休克蛋白(Heat Shock Proteins,HSPs)是一种分子伴侣, 具有广泛的生物学活性。从香鱼(Plecoglossus altivelis)肝cDNA文库获得HSP90 β亚型(PaHSP90β) cDNA全序列。PaHSP90β全长2726 bp, 包含1个由2175 bp组成的开放阅读框, 编码724个氨基酸, 预测相对分子质量为83.30 ku, 等电点为4.83。氨基酸序列分析显示, PaHSP90β含有HSP90的5个保守信号区, 氨基酸序列第34~188处具有HATPase (histidine kinase-like ATPases)结构域; 与白鲑(Coregonus lavaretus) HSP90β亲缘关系最近, 氨基酸序列同源性高达95.8%。实时荧光定量PCR(Quantitative real-time PCR, qPCR)结果表明, PaHSP90β mRNA主要在香鱼中表达量最高, 脾脏次之。镉胁迫后, 香鱼肝、脾、肾、鳃和肠组织中PaHSP90β mRNA表达显著上调; 盐度胁迫后, 除肠组织外, 其他组织中PaHSP90β mRNA表达总体显著上调。综上, PaHSP90β与重金属及盐度胁迫密切相关, 可能参与鱼类抵抗环境胁迫的生理过程。

关键词: 香鱼, HSP90β, 镉胁迫, 盐度胁迫, 实时荧光定量PCR

Abstract: Heat shock proteins (HSPs) are one type of molecular chaperones, and have a wide range of biological activities. The full-length cDNA sequence of ayu HSP90β gene (PaHSP90β) was obtained from the liver tissue cDNA library of ayu (Plecoglossus altivelis). The cDNA sequence of PaHSP90β was 2726 nucleotides, containing a 2175 bp open reading frame that encoded a protein of 724 amino acids with a deduced molecular weight (MW) of 83.30 ku and a isoelectric point (pI) of 4.83. Multiple sequence alignment of complete amino acid sequences showed that PaHSP90β contained five conserved HSP90 family signatures, and a typical HATPase (histidine kinase-like ATPases) domain, which was located at positions 34 to 188. PaHSP90β shared the highest amino acid sequence identity (95.8%) with that of European whitefish (Coregonus lavaretus). Quantitative real-time PCR (qPCR) analysis showed that PaHSP90β mRNA had the highest expressional level in ayu liver, followed by the spleen. Upon Cd exposure, PaHSP90β transcripts were significantly up-regulated in liver, spleen, kidney, intestine and gill of ayu. After the salinity stress, PaHSP90β mRNA expression was significantly up-regulated in all the tissues except the intestine. In summary, PaHSP90β had a close relationship with heavy metal and salinity stress, and may be involved in physiological processes against environmental stresses in fish.
   

Key words: Plecoglossus altivelis, HSP90β, cadmium stress, salinity stress, quantitative real-time PCR