水产学报2017，Vol.41Issue(4)：481-489,9.DOI:0.11964/jfc.20160410364
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
摘要
Abstract
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 ofL.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.关键词
大黄鱼/低温胁迫/冷诱导结合蛋白(CIRP)/组织表达Key words
Larimichthys crocea/cold treatment/cold inducible RNA-binding protein (CIRP)/tissue expression分类
生物科学引用本文复制引用
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):481-489,9.基金项目
国家“八六三”高技术研究发展计划(2012AA10A403-4) （2012AA10A403-4）
浙江省重大科技专项(2016C02055-7)National High Technology Research and Development Program of China (863 Program)(2012AA10A403-4) （2016C02055-7）
Key Technologies R&D Project of Zhejiang Province (2016C02055-7) （2016C02055-7）
