| 宽体沙鳅(Botia reevesae)热休克蛋白70 基因的克隆及表达分析 |
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| 引用本文： | 覃川杰,顾顺樟,赵大显,陈立侨,李二超,齐泽民. 宽体沙鳅(Botia reevesae)热休克蛋白70 基因的克隆及表达分析[J]. 海洋与湖沼, 2013, 44(6): 1584-1591 |
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| 作者姓名： | 覃川杰 顾顺樟 赵大显 陈立侨 李二超 齐泽民 |
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| 作者单位： | 内江师范学院生命科学学院 内江,南昌大学生命科学与食品工程学院,华东师范大学生命科学学院 上海,华东师范大学生命科学学院 上海,内江师范学院生命科学学院 内江,内江师范学院生命科学学院 内江 |
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| 基金项目： | 四川省教育厅项目, 11ZB025 号; 四川省科技厅项目, 2011NZ0075 号; 国家科技支撑计划课题资助项目, 2012BAD25B03号; 国家自然科学基金项目, 31260642 号; 江西省自然(青年)科学基金, 20132BAB214015 号; 中国博士后科学基金第六批特别资助, 2013T60650 号; 内江师范学院大学生科研立项, 13NSD-77 号。 |
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| 摘 要： | 为深入分析热休克蛋白响应胁迫的分子机制,实验以宽体沙鳅(Botia reevesae)为研究对象,利用同源克隆和cDNA 末端快速扩增(rapid amplification of cDNA ends,RACE) 技术克隆得到宽体沙鳅热休克蛋白70(BR-HSP70) 的cDNA 全长。结果发现,BR-SP70 cDNA全长为2,371bp,包含1,947 bp的开放阅读框(opening reading frame,ORF),102 bp 5’-非编码区(untranslated region,UTR)和322 bp 3’-UTR等。通过序列同源性比对发现,BR-HSP70 cDNA与团头鲂(Megalobrama amblycephala)和猪(Sus scrofa)的同源性分别为98%及83%,且ORF编码的649个氨基酸中含有HSP70家族的家族信号标签、N-糖基化位点及EEVD等能位点等保守序列。上述结果表明,本研究所获的基因为宽体沙鳅 HSP70基因。实时荧光定量PCR 分析发现,氨氮胁迫和嗜水气单胞菌(Aeromonas hydrophila)侵染均会显著上调宽体沙鳅鳃、肝脏及肾脏HSP70 mRNA的表达,表明HSP70基因在宽体沙鳅应对环境胁迫中发挥了重要的抗应激作用。
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| 关 键 词： | 宽体沙鳅、热休克蛋白70、氨氮、嗜水气单胞菌、胁迫 |
| 收稿时间： | 2013-05-27 |
| 修稿时间： | 2013-07-29 |
THE CLONING AND EXPRESSION OF HEAT SHOCK PROTEIN 70 cDNA IN BOTIA REEVESAE |
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| QIN Chuan-Jie,GU Shun-Zhang,ZHAO Da-Xian,CHEN Li-Qiao,LI Er-Chao and QI Ze-Min. THE CLONING AND EXPRESSION OF HEAT SHOCK PROTEIN 70 cDNA IN BOTIA REEVESAE[J]. Oceanologia Et Limnologia Sinica, 2013, 44(6): 1584-1591 |
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| Authors: | QIN Chuan-Jie GU Shun-Zhang ZHAO Da-Xian CHEN Li-Qiao LI Er-Chao QI Ze-Min |
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| Affiliation: | School of Life Science, Neijiang Normal University,School of Life Science and Food Engineering, Nanchang University,College of Life Science, East China Normal University,College of Life Science, East China Normal University,School of Life Science, Neijiang Normal University,School of Life Science, Neijiang Normal University |
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| Abstract: | We investigated the effect of pathogenic bacterial challenge and acute sublethal ammonia-N exposure on heat shock protein 70 (HSP70) expression in Botia reevesae. Results show that the HSP70 cDNA in B. reevesaeis 2371bp, containing an open reading frame at 1947bp, a 102bp 5'-untranslated region (UTR), a 322bp 3'-UTR, and five common putative functional domains. By sequence comparisons, we found that the deduced amino acid sequence of the HSP70 had an overall identity of 98%, and 83% to that of Megalobrama amblycephala andSus scrofa, respectively. Real time quantitative PCR analysis showed the HSP70 mRNA expression levels was significantly (P<0.05) up-regulated in gill, liver, spleen, and kidney by Aeromonas hydrophilachallenge or under ammonia-N exposure. It is believed that B. reevesaeHSP70 is involved in the resistance to pathogenic bacteria stress. |
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| Keywords: | Botia reevesae,Heat shock protein 70,Ammonia-N,Aeromonas hydrophila,Stress |
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