| 摘要: |
| 为鲆鲽鱼耐低温标记辅助育种研究和应用提供候选基因, 并了解耐寒相关基因在低温下的表达图示, 作者通过同源克隆以及5?/3?RACE 的方法扩增得到了牙鲆(Paralichthys olivaceus）CIRP和HMGB1基因。组织表达谱分析显示, 这两个基因在脑、鳃、心脏、肌肉、肝脏、肠等组织均有表达, 但HMGB1基因在鳃中表达较弱。低温胁迫后, 两基因随着胁迫时间的延长其表达量在肌肉中都出现先上升后下降的趋势: 在0～12 h 逐渐上升并达到最大值, 然后下降恢复至最初水平。但在肝脏和脑组织中的变化趋势却不同, 低温胁迫后0～2 hHMGB1基因在脑中的表达量上升至最高, 然后下降至最初水平,在肝脏中变化不大。而CIRP基因在低温胁迫后0～2 h 在肝脏中表达量处于上升趋势, 然后下降至初始水平, 在脑中变化不大。由此推测, 这两个基因在低温条件下都参与了机体对外界的抵抗, 特别是在抵抗外界寒冷的主要组织—肌肉中发挥作用, 可能共同参与了机体的免疫反应, 对低温胁迫环境下的鱼体进行保护, 可以作为鱼类耐低温标记筛选的候选基因。 |
| 关键词: 牙鲆(Paralichthys olivaceus) 低温胁迫 CIRP和HMGB1 克隆 RT-PCR |
| DOI：10.11759/hykx20140304001 |
| 分类号: |
| 基金项目:国家863 计划资助项目(2012AA10A408, 2012AA092203);国家973 计划资助项目(2010CB126304); 国家科技基础条件平台建设运行项目(2006DKA30470017) |
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| Cloning and expression analysis of cold-tolerance related genes, CIRP and HMGB1, in Paralichthys olivaceus |
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| Abstract: |
| In order to learn the expression patterns of CIRP and HMGB1 genes and provide more candidate genes for cold-resistant marker assistance breeding of flatfish, these two cold-tolerance related genes in olive flounder (Paralichthys olivaceus) were cloned using 5?and 3? RACE methods. The open reading frames of flounder CIRP and HMGB1 were 932 bp (GenBank accession number, KC735176) and 1222 bp (GenBank accession number, KC422724), respectively. The expression patterns of those two genes in flounder tissues were studied using RT-PCR analysis. The expression levels of HMGB1 were higher in liver, heart, muscle and, intestines than those in brain and gill. Under the low water temperature, expression levels of CIRP and HMGB1 genes in flounder brain, liver and muscle tissues were also studied using real-time RT-PCR method. In muscle, the expression levels of CIRP and HMGB1 increased during 0～12 h, and then decreased with the prolonging of treatment time. The expression level of HMGB1 in brain tissue increased during 0～2 h, and then decreased to the initial level, and there was no difference in expression levels in the liver tissue during the whole cold treatment process. The expression levels of CIRP in liver tissue increased during 0～2 h, and then decreased to the initial level. But there was no obvious fluctuation in brain tissue. According to the above results, we speculate that CIRP and HMGB1 could participate in flounder cold toleration nd may promote inflammatory responses under low temperatures, so both genes could be candidate genes for cold-resistant molecular marker selection of flounder. |
| Key words: Paralichthys olivaceus cold treatment CIRP and HMGB1 RT-PCR |
