唐鱼Caspase-3和Caspase-9 cDNA全长克隆及胁迫表达分析  

CLONING AND EXPRESSION OF WHITE CLOUD MOUNTAIN MINNOW(TANICHTHYS ALBONUBES) CASPASE-3 AND CASPASE-9 cDNAs

作　　者：徐胜威[1] 李恬[1] 吴金英[1] 梁健宏 刘汉生 连常平 

机构地区：[1]中山大学生命科学学院,水生经济动物研究所暨广东省水生经济动物良种繁育重点实验室,广州510275 [2]广州市海洋与渔业自然保护区管理站,广州510405

出　　处：《水生生物学报》2011年第1期138-144,共7页Acta Hydrobiologica Sinica

基　　金：国家人才基金(基地)项目(J0730638);广州市海洋与渔业自然保护区管理站基金项目(2007-33000-4206649)资助

摘　　要：研究以唐鱼(Tanichthys albonubes)鳃部组织为材料,利用同源克隆和RACE方法,首次克隆得到唐鱼Caspase-3和Caspase-9 cDNA全长序列,并对其生物信息学进行了分析和鉴定。实验结果表明唐鱼Caspase-3 cDNA全长2153 bp,开放读码框(Open Reading Frame,ORF)为840 bp,编码一段长为279个氨基酸的多肽序列;Caspase-9 cDNA全长为1758 bp,ORF为1323 bp,编码440个氨基酸。生物信息学分析表明Caspase-3和Caspase-9与其他鱼类在蛋白结构上较为保守,在重要的功能域呈现高度的保守性。应用实时荧光定量PCR技术,分析了氯氰菊酯对唐鱼鳃部组织表达Caspase-3和Caspase-9的影响,在氯氰菊酯的胁迫下,Caspase-3和Caspase-9的表达呈现抑制状态,且呈现明显的浓度和时间依赖效应。The white cloud mountain minnow（Tanichthys albonubes） is an important aquarium fish,and also a potential experimental model organism.In order to get the knowledge of apoptosis under environmental stress in this species,we have firstly cloned and sequenced the full-length cDNAs of Caspase-3 and Caspase-9 from the gill tissue of T.albo-nubes using homology cloning approach combined with RACE method.In this study,the bioinformatics analysis of Caspase-3 and Capase-9 full-length cDNA sequences were presented.The results indicated that the full-length cDNA of Caspase-3 was composed of 2153 base pairs（bp）,which included a 840 bp open reading frame（ORF） encoding a 279-amino acid peptide,whereas the full-length cDNA of Caspase-9 was composed of 1758 bp,which included a 1323 bp ORF encoding a 440-amino acid peptide.The identity comparison and the phylogenetic analysis showed that the predicted Caspase-3 and Caspase-9 proteins of T.albonubes were structurally similar to those of other fish,with high conservation in important functional domains.Real-time PCR analysis showed that the expression levels of Caspase-3 and Caspase-9,under cypermethrin-induced stress conditions,were suppressed compared to control group,and that Caspase-3 and Caspase-9 mRNA levels were significantly affected in a dose-and time-dependent manner by cyperme-thrin.These results suggest that cypermethrin plays a role in the suppression of apoptosis of T.albonubes,and that the anti-apoptotic effect of cypermethrin may be beneficial to this fish so that it can survive in harsh environments.

关 键 词：唐鱼 CASPASE-3 CASPASE-9 克隆 氯氰菊酯 表达 

分 类 号：Q781[生物学—分子生物学]