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HONG Jingni,MAO Yong,NIU Sufang,SUN Tiantian,SU Yongquan. 2015. Molecular characterization and expression of HSP70, HSF and HSBP genes in Octopus vulgaris during thermal stress. Acta Oceanologica Sinica, 34(8):62-72
Molecular characterization and expression of HSP70, HSF and HSBP genes in Octopus vulgaris during thermal stress
真蛸HSP70、HSF和HSBP基因克隆、序列分析及热应激下表达分析
Received:June 08, 2014  Revised:October 28, 2014
DOI:10.1007/s13131-015-0708-6
Key words:Octopus vulgaris  high temperature stress  HSP70  HSF  HSBP
中文关键词:  真蛸  高温应激  热休克蛋白70  热休克转录因子  热休克转录因子结合蛋白
基金项目:The Public Science and Technology Research Funds Projects of Ocean under contract No. 201005013.
Author NameAffiliationE-mail
HONG Jingni College of Ocean and Earth Sciences, Xiamen University, Xiamen 361102, China  
MAO Yong College of Ocean and Earth Sciences, Xiamen University, Xiamen 361102, China  
NIU Sufang College of Ocean and Earth Sciences, Xiamen University, Xiamen 361102, China  
SUN Tiantian College of Ocean and Earth Sciences, Xiamen University, Xiamen 361102, China  
SU Yongquan College of Ocean and Earth Sciences, Xiamen University, Xiamen 361102, China yqsu@xmu.edu.cn 
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Abstract:
      Temperature is an important environmental factor that affects the growth and survival of Octopus vulgaris, the common octopus. To understand the protective mechanism that O. vulgaris exhibits under heat stress, we used rapid amplification of cDNA ends (RACE) to obtain full-length sequences of three heat stress response related genes: (1) the heat shock protein 70 (OvHSP70), (2) the heat shock transcription factor (OvHSF) , and (3) the heat shock factor-binding protein (OvHSBP) of O. vulgaris. The OvHSP70, OvHSF, and OvHSBP proteins contained 2 222 bp, 2 264 bp, 841 bp that encoded for 635, 458 and 90 amino acids, respectively. The results of multiple sequence alignment showed that the amino acid sequences of OvHSP70 were highly conserved with respect to other species. Similarly, the DNA binding domain, the trimerization domain of OvHSF, and the coiled coil region of OvHSBP also had highly conserved regions. The real-time polymerase chain reaction (PCR) results indicated that OvHSP70 was temperature-dependent and time-dependent, showing a positive response to heat stress. On exposure to 28℃ and to 30℃, the mRNA expression levels of OvHSF and OvHSBP were higher than those in the control group at 24℃. The mRNA expression of OvHSBP significantly increased with heat treatment at 26℃, while the mRNA expression of OvHSF decreased. The experimental results indicated that the expression of OvHSP70, OvHSF and OvHSBP were all sensitive to heat stress, which suggests that these three genes may play an important role for O. vulgaris in responding to environmental stress. Thus, this study sets a theoretical foundation for further in-depth studies on the molecular protective mechanisms of the heat response in O. vulgaris.
中文摘要:
      温度是影响真蛸生长生存的重要环境因子,为了了解真蛸面对热应激的分子保护机制,我们利用cDNA末端快速克隆(RACE)技术获得了真蛸热休克蛋白70(OvHSP70)、热休克转录因子(OvHSF)和热休克转录因子结合蛋白(OvHSBP)等3个热应激相关基因的cDNA全长,3个基因全长分别为2222 bp、2264bp、841bp,编码635、458和90个氨基酸.多序列比对结果显示,OvHSP70氨基酸序列与其他物种具有高保守性,同时,OvHSF的DBD区域和三聚区域和OvHSBP的螺旋卷曲区域同源性较高.实时荧光定量PCR结果表明,OvHsp70基因mRNA表达量具有明显的温度和时间依赖性,展现了该基因对高温应激的积极响应.不同温度组实验中,在28℃和30℃组,OvHSF和OvHSBP的mRNA表达量显著高于常温组(24℃);而在26℃组,OvHSBP mRNA表达量显著上升,OvHSF的mRNA表达量则下降.本实验结果显示,OvHSP70、OvHSF和 OvHSBP 3个基因对高温应激都有不同程度的响应,预示着这3个基因对真蛸响应环境压力起到了重要的作用,为将来进一步研究真蛸热应激分子机制奠定理论基础.
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