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	赤点石斑鱼HSP60基因克隆及弧菌应激前后的组织表达特性分析; Cloning of HSP60 gene from Epinephelus akaara and its express characterization before and after vibrionic stressed
	曲朦 ; 施晓峰 ; 张之文 ; 丁少雄
	2011
关键词	赤点石斑鱼 HSP60 克隆 组织表达特性 Epinephelus akaara heat shock protein60 cloning express characterization
英文摘要	HSP60(HEAT SHOCk PrOTEIn 60,热休克蛋白60)作为高度保守的热休克蛋白家族的一员,不仅可以在应激状态下帮助其他蛋白正确折叠并恢复天然构象,还可作为危险信号作用于天然免疫系统。基于HSP60的EST序列设计特异引物,采用rACE技术成功克隆获得赤点石斑鱼HSP60 CdnA全序列及基因组全序列。其CdnA序列全长2 351 bP,包括75 bP的5′uTr,539 bP的3′uTr和1 737 bP的开放阅读框,共编码578个氨基酸。核苷酸及氨基酸序列比对结果显示该基因与其他脊椎动物的相似性很高。基于氨基酸序列构建的系统进化树与传统物种树基本吻合,说明HSP60可能是1个潜在的可用于物种系统进化研究的良好标记。所获得的赤点石斑鱼HSP60基因组全序列含有8个内含子和9个外显子,也与已知其他硬骨鱼类基本一致。这种在序列特征、蛋白空间结构及基因组结构上的高度保守性意味着HSP60在动物生命活动中可能发挥着至关重要的作用。通过实时PCr检测结果发现HSP60在赤点石斑鱼各组织中均有表达,而攻毒后鱼体肝的表达量显著增高(P<0.05),鳃、脾、胃和脑等重要组织器官的表达量也有不同程度的提高,该结果证实了HSP60基因在鱼类非特异性免疫应答中发挥了作用。; As a major member of heat shock protein families,HSP60 can not only be served as a molecular chaperone to promote protein disaggregation and proper refolding,but also be regarded as a threatened signal under stress to elicit a potent proinflammatory response in cells of the innate immune system.Based on the EST sequences from cDNA library,the full length cDNA of HSP60 from E.akaara was successfully obtained by RACE technique.The full-length of HSP60 is 2351bp,including 75bp of 5′UTR,539bp of 3′UTR,and a 1737bp open reading frame encoding a protein of 578 amino acids.The alignment result shows that the nuclear acid sequence and amino acids sequence of HSP60 from E.akaara share high identity with other vertebrate homologs.The phylogenetic tree based on the HSP60 amino acid sequences is congruent with their evolutionary relationships,which indicates that HSP60 may be an appropriate marker for the phylogenetic research among interspecies.The E.akaara HSP60 gene contains 9 introns and 10 exons,the same as other teleosteans.The conserved sequence,gene structure and 3D protein structure show that HSP60 may play an important role in life activities.Real-time PCR was also conducted to detect the express characterization of the E.akaara HSP60 gene.The result shows that HSP60 can be detected in various tissues,and after Vibrio harveyi-challenging,the expression of HSP60 in the liver increases dramatically,most of other tissues are also detected increasing to a great extent,which supports that HSP60 maybe play a role in fish innate immune response.; 福建省自然科学基金(2060203);福建省新世纪人才资助计划
语种	zh_CN
内容类型	期刊论文
源URL	[http://dspace.xmu.edu.cn/handle/2288/104329]
专题	海洋环境－已发表论文
推荐引用方式 GB/T 7714	曲朦,施晓峰,张之文,等. 赤点石斑鱼HSP60基因克隆及弧菌应激前后的组织表达特性分析, Cloning of HSP60 gene from Epinephelus akaara and its express characterization before and after vibrionic stressed[J],2011.
APA	曲朦,施晓峰,张之文,&丁少雄.(2011).赤点石斑鱼HSP60基因克隆及弧菌应激前后的组织表达特性分析..
MLA	曲朦,et al."赤点石斑鱼HSP60基因克隆及弧菌应激前后的组织表达特性分析".(2011).

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