文章摘要
苗亮,李明云,陈莹莹,胡谋,陈炯.大黄鱼冷诱导结合蛋白(CIRP)基因cDNA克隆及低温胁迫对其时空表达的影响[J].水产学报,2017,41(4):481~489
大黄鱼冷诱导结合蛋白(CIRP)基因cDNA克隆及低温胁迫对其时空表达的影响
Cloning of cold inducible RNA-binding protein (CIRP) gene in Larimichthys crocea and its expression analysis under cold treatments
投稿时间:2016-04-15  修订日期:2016-08-06
DOI:10.11964/jfc.20160410364
中文关键词: 大黄鱼  低温胁迫  冷诱导结合蛋白(CIRP)  组织表达
英文关键词: Larimichthys crocea  cold treatment  cold inducible RNA-binding protein (CIRP)  tissue expression
基金项目:国家“八六三”高技术研究发展计划(2012AA10A403-4);浙江省重大科技专项(2016C02055-7)
作者单位E-mail
苗亮 宁波大学教育部应用海洋生物技术重点实验室, 浙江 宁波 315211  
李明云 宁波大学教育部应用海洋生物技术重点实验室, 浙江 宁波 315211 limingyun@nbu.edu.cn 
陈莹莹 宁波大学教育部应用海洋生物技术重点实验室, 浙江 宁波 315211  
胡谋 宁波大学教育部应用海洋生物技术重点实验室, 浙江 宁波 315211  
陈炯 宁波大学教育部应用海洋生物技术重点实验室, 浙江 宁波 315211  
摘要点击次数: 1473
全文下载次数: 933
中文摘要:
      为研究冷诱导结合蛋白(CIRP)在大黄鱼低温适应过程中的作用,采用同源克隆技术获得了大黄鱼CIRP基因,并通过实时荧光定量PCR (qRT-PCR)技术检测了低温胁迫下大黄鱼各组织中CIRP基因的表达变化。结果显示,大黄鱼CIRP基因cDNA序列全长1211 bp,推测编码一个由182个氨基酸组成、分子量为19 ku的蛋白。序列比对显示硬骨鱼类CIRP基因序列较为保守,特别是N端的RNA识别基序(RRM)高度保守。系统进化树分析显示大黄鱼CIRP与银鳕相似性最高(81.5%),所有硬骨鱼类聚为一个大支。慢性低温胁迫中(12℃缓降至6℃),皮肤和肌肉中CIRP基因的表达量呈随温度降低而升高的趋势,6℃时表达量分别为12℃时的11.56倍和9.03倍;鳃、脑和心脏中CIRP基因表达量均呈先显著上升后显著下降的趋势,其峰值均出现在8℃时,表达量分别为12℃时的5.07、7.69和2.83倍;肠、肾脏、肝脏和脾脏中的表达量随温度降低而下降,6℃时的表达量最低,与12℃时相比降幅分别为80.0%、65.6%、91.2%、55.6%。急性低温胁迫(由12℃骤降至8℃并胁迫4 h)条件下,鳃、皮肤、脑、肝脏、心脏中CIRP基因表达量均呈先升高后降低的变化,反映了机体对低温胁迫的应激、适应过程;肌肉和肠中的表达量则始终显著低于胁迫前。慢性和急性低温胁迫中各组织CIRP表达量变化的差异提示,大黄鱼机体在低温胁迫响应和适应中有着多种调节机制。研究表明,CIRP基因参与了大黄鱼应对低温胁迫的过程。
英文摘要:
      In order to investigate the function of cold inducible RNA-binding protein (CIRP) of large yellow croaker Larimichthys crocea in cold acclimatization, CIRP gene was cloned and the expression changes were analyzed by real-time quantitative PCR (qRT-PCR). The full-length of cDNA sequence of CIRP is 1211 bp, encodes 182 amino acid residues and the protein molecular weight is 19 ku. Amino acid sequence alignment showed that CIRP was conserved in teleost, especially the RNA recognition motif (RRM) at N-terminal was highly conserved. The phylogenetic tree analysis showed that CIRP of L. crocea had the highest identity to sablefish Anoplopoma fimbria (81.5%), CIRP of L. crocea and other teleost formed a clade. During chronic cold stress (water temperature slowly decreased from 12℃ to 6℃), the CIRP expression of skin and muscle significantly increased with the temperature decreasing, in which the expression levels at 6℃ rose 11.56 and 9.03 times compared to 12℃. In gill, brain and heart, the expression of CIRP increased at first and then decreased, and the maximum appeared at 8℃ which were 5.07, 7.69 and 2.83 times compared to 12℃. The expression level significantly decreased with the temperature increasing in intestine, kidney, liver and spleen, and the decreasing amplitude at 6℃ were 80.0%, 65.6%, 91.2% and 55.6% compared to 12℃ respectively. During acute cold stress (from 12℃ to 8℃ immediately and maintained for 4 h), the expression levels of CIRP in gill, skin, brain, liver and heart increased first and then decreased, reflected the stress and adaptation process of L. crocea to cold. Compared to 12℃, the CIRP expression in muscle and intestine were always at a lower level during acute cold stress. The difference of tissue expression of CIRP between chronic and acute cold stresses suggested that there were multiple regulation mechanisms replying to cold stress in L. crocea. The present study indicated CIRP was involved in L. crocea responding to cold stress, and the results could provide fundamental data for further research on the mechanism of response to low temperature in L. crocea.
HTML   查看全文   下载PDF阅读器
关闭

手机扫一扫看