College of Marine Sciences, South China Agricultural University, Guangzhou, Guangdong, 510642, China.
Qingyuan North River Fishery Science Institute, Qingyuan, Guangdong, 511510, China.
Cell Stress Chaperones. 2018 Jul;23(4):791-796. doi: 10.1007/s12192-017-0869-7. Epub 2018 Mar 14.
The great albino northern snakehead, Channa argus, is one of the most important economical fish in China. In the present study, cDNA encoding heat shock protein 60 (HSP60) was cloned and characterized. The cDNA was 2462 bp, containing an open reading frame (ORF) encoding a 575-amino-acids polypeptide of 61.10 kDa (theoretical isoelectric point [pI]: 5.66). BLAST analysis showed that AcaHSP60 was highly similar with other HSP60s, and three conserved amino acid blocks and characteristic motifs or domains defined as HSP60 protein family signatures. Genomic DNA analysis showed that AcaHSP60 had ten exons in the coding region (from 94 to 336 bp). Changes in AcaHSP60 gene expression profiles in albino C. argus experimentally exposed to different temperature stress (8.5, 26, and 37 °C) was investigated. Quantitative real-time PCR and western blot analysis revealed that tissue-specific AcaHSP60 expressions were in the spleen, muscle, liver, kidney, heart and brain. Expression was highly significantly stimulated after heat shock (37 °C), but showed no significant differences after cold treatment (8.5 °C) except in the brain. In summary, these results showed that AcaHSP60 was significantly tissue specific and indicate that AcaHSP60 expression might be sensitive to thermal resistance in albino C. argus.
白化北方蛇头鱼,又称中国团头鲂,是中国最重要的经济型鱼类之一。本研究克隆并鉴定了热休克蛋白 60(HSP60)的 cDNA。该 cDNA 长 2462bp,含有一个开放阅读框(ORF),编码一个 575 个氨基酸的多肽,分子量为 61.10kDa(理论等电点 [pI]:5.66)。BLAST 分析表明,AcaHSP60 与其他 HSP60s 高度相似,并且具有三个保守的氨基酸块和特征基序或定义 HSP60 蛋白家族特征的结构域。基因组 DNA 分析表明,AcaHSP60 在编码区有十个外显子(94-336bp)。研究了白化北方蛇头鱼在不同温度应激(8.5、26 和 37°C)下 AcaHSP60 基因表达谱的变化。定量实时 PCR 和 Western blot 分析显示,AcaHSP60 在组织中的表达具有特异性,分别在脾脏、肌肉、肝脏、肾脏、心脏和大脑中表达。热激(37°C)后 AcaHSP60 的表达显著上调,但冷处理(8.5°C)后除大脑外无显著差异。总之,这些结果表明 AcaHSP60 具有明显的组织特异性,表明 AcaHSP60 的表达可能对白化北方蛇头鱼的耐热性敏感。