| 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(heat shock protein, BR-HSP70)的cDNA全长。结果发现, BR-HSP70 cDNA 全长为2371bp, 包含1947bp的开放阅读框(opening reading frame, ORF), 102bp 5′-非编码区(untranslated region, UTR)和322bp 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等保守区域。结果表明, 克隆的cDNA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(P<0.05), 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。 |
| 关键词: 宽体沙鳅 热休克蛋白70 氨氮 嗜水气单胞菌 胁迫 |
| DOI：10.11693/hyhz20130700097 |
| 分类号: |
| 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。 |
附件 |
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| THE CLONING AND EXPRESSION OF HEAT SHOCK PROTEIN 70 cDNA IN BOTIA REEVESAE |
|
QIN Chuan-Jie,GU Shun-Zhang,ZHAO Da-Xian,CHEN Li-Qiao,LI Er-Chao,QI Ze-Min
|
| 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. reevesae
HSP70 is involved in the resistance to pathogenic bacteria stress. |
| Key words: Botia reevesae heat shock protein 70 ammonia-N Aeromonas hydrophila stress |
