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c DNA文库获得HSP90β亚型(Pa HSP90β)c DNA全序列。Pa HSP90β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,Pa HSP90β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,q PCR)结果表明,Pa HSP90βmRNA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Pa HSP90βmRNA表达显著上调;盐度胁迫后,除肠组织外,其他组织中Pa HSP90βmRNA表达总体显著上调。综上,Pa HSP90β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。
Heat shock proteins( HSPs) are one type of molecular chaperones,and have a wide range of biological activities. The fulllength c DNA sequence of ayu HSP90β gene( Pa HSP90β) was obtained from the liver tissue c DNA library of ayu( Plecoglossus altivelis). The c DNA sequence of Pa HSP90β 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( p I) of 4. 83. Multiple sequence alignment of complete amino acid sequences showed that Pa HSP90β contained five conserved HSP90 family signatures,and a typical HATPase( histidine kinase-like ATPases) domain,which was located at positions 34 to 188. Pa HSP90β shared the highest amino acid sequence identity( 95. 8%) with that of European whitefish( Coregonus lavaretus). Quantitative real-time PCR( q PCR) analysis showed that Pa HSP90β mRNA had the highest expressional level in ayu liver,followed by the spleen. Upon Cd exposure,Pa HSP90β transcripts were significantly up-regulated in liver,spleen,kidney,intestine and gill of ayu. After the salinity stress,Pa HSP90β mRNA expression was significantly up-regulated in all the tissues except the intestine. In summary,Pa HSP90β had a close relationship with heavy metal and salinity stress,and may be involved in physiological processes against environmental stresses in fish.
引文
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