Key Laboratory for Sustainable Utilization of Marine Fisheries Resources, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao, PR China.
Fish Shellfish Immunol. 2012 Jun;32(6):1191-7. doi: 10.1016/j.fsi.2012.03.008. Epub 2012 Mar 13.
Heat shock protein 90 (HSP90) is a highly conserved molecular chaperone contributing to the folding, maintenance of structural integrity and proper regulation of a subset of cytosolic proteins. In this study, a heat shock protein 90 cDNA named EcHSP90 was cloned from the hepatopancreas of ridgetail white prawn Exopalaemon carinicauda by reverse transcription polymerase chain reaction (RT-PCR) coupled with rapid amplification of cDNA ends (RACE) approaches. The full-length cDNA of EcHSP90 was of 2695 bp, including an open reading frame (ORF) of 2163 bp encoding a polypeptide of 720 amino acids with an estimated molecular mass of 82.73 kDa and an estimated isoelectric point of 4.83. BLAST analysis revealed that the EcHSP90 shared high similarity (87.6%-75.24%) with other known HSP90s. The five conserved amino acid blocks defined as HSP90 protein family signatures were also identified in EcHSP90, which indicated that EcHSP90 should be a cytosolic member of the HSP90 family. Quantitative real-time RT-PCR analysis revealed that EcHSP90 transcript could be detected in all the tested tissues, and strongly expressed in ovary of E. carinicauda. The transcript of EcHSP90 in hepatopancreas of E. carinicauda showed different expression profiles after pH and ammonia-N stresses. The results indicated that EcHSP90 was a constitutive and inducible expressed protein and could be induced by various stresses from environment.
热休克蛋白 90（HSP90）是一种高度保守的分子伴侣，有助于折叠、维持结构完整性和正确调节细胞质蛋白的一部分。在这项研究中，通过反转录聚合酶链反应（RT-PCR）与快速扩增 cDNA 末端（RACE）方法，从脊尾白对虾 Exopalaemon carinicauda 的肝胰腺中克隆出一种热休克蛋白 90 cDNA，命名为 EcHSP90。EcHSP90 的全长 cDNA 为 2695bp，包括一个 2163bp 的开放阅读框（ORF），编码一个 720 个氨基酸的多肽，估计分子量为 82.73kDa，等电点为 4.83。BLAST 分析表明，EcHSP90 与其他已知的 HSP90s 具有高度相似性（87.6%-75.24%）。在 EcHSP90 中还鉴定出了五个保守的氨基酸块，定义为 HSP90 蛋白家族特征，这表明 EcHSP90 应该是 HSP90 家族的细胞质成员。定量实时 RT-PCR 分析显示，EcHSP90 转录本可在所有检测到的组织中检测到，并在脊尾白对虾的卵巢中强烈表达。脊尾白对虾肝胰腺中 EcHSP90 的转录本在 pH 和氨氮胁迫后表现出不同的表达谱。结果表明，EcHSP90 是一种组成型和诱导型表达蛋白，可以被环境中的各种应激诱导。
