中华鲟热休克蛋白hsp30基因cDNA的克隆及其在高温胁迫下的表达分析  

Molecular cloning and expression analysis under high-temperature stress of heat shock protein hsp30 gene cDNA from Acipenser sinensis

作　　者：司凯歌 江南[2] 张海耿[3] 周勇[2] 刘文枝[2] 曾令兵 倪琦[3] 范玉顶[2] SI Kai-ge;JIANG Nan;ZHANG Hai-geng;ZHOU Yong;LIU Wen-zhi;ZENG Ling-bing;NI Qi;FAN Yu-ding(National Demonstration Center for Experimental Fisheries Science Education(Shanghai Ocean University),Shanghai 201306,China;Yangtze River Fisheries Research Institute,Chinese Academy of Fishery Sciences,Wuhan 430223,China;Fishery Machinery and Instrument Research Institute,Chinese Academy of Fishery Sciences,Shanghai 200092,China)

机构地区：[1]水产科学国家级实验教学示范中心(上海海洋大学),上海201306 [2]中国水产科学研究院长江水产研究所,武汉430223 [3]中国水产科学研究院渔业机械仪器研究所,上海200092

出　　处：《淡水渔业》2019年第6期20-26,共7页Freshwater Fisheries

基　　金：国家重点基础研究发展计划(973计划)项目(2015CB150703)

摘　　要：实验克隆了中华鲟(Acipenser sinensis)热休克蛋白hsp 30基因cDNA的全长、分析了其分子结构与特征,并研究了其在高温胁迫下的表达水平。结果显示,中华鲟hsp 30基因cDNA序列全长为1037 bp,其中开放阅读框(ORF)636 bp,5′端非编码区(5′UTR)38 bp,3′端非编码区(3′UTR)363 bp,共编码211个氨基酸。氨基酸多序列比对发现含有一个保守的α晶状体结构;系统进化分析显示,中华鲟HSP30与鱼类HSP30聚为一支,与小体鲟HSP30氨基酸序列相似性最高,为79%。荧光定量PCR结果表明,中华鲟hsp 30基因在皮肤中的表达量最高,肝脏次之,在肠中的表达量最低。高温胁迫后,心脏、脾脏、肾脏和皮肤中hsp 30基因表达量均显著增加,表明这些器官在中华鲟应对高温胁迫中可能起着重要作用。The full length of heat shock protein hsp 30 gene cDNA of Acipenser sinensis was cloned and the molecular structure and characteristics were analysed in this study.Furthermore,the expression levels under high-temperature stress were studied.The results showed that the full-length cDNA sequences of hsp 30 gene were 1037 bp,with a 636 bp open reading frame(ORF)which encoded the protein with 211 amino acids as well as 38 bp 5′untranslated region(5′UTR)and 363 bp 3′untranslated region(3′UTR).Amino acid multiple sequence alignments revealed that HSP30 had a conservedα-crystal structure.Phylogenetic analysis showed that the HSP30 amino acid sequence of A.ruthenus was clustered together with fish,and also showed the highest similarity(79%)with the A.ruthenus.The results of real-time PCR showed that hsp 30 extensively expressed in various tissues.The highest expression was detected in the skin,followed by the liver,and the lowest was in the intestine.After high-temperature stimulation,the expression of hsp 30 in the heart,spleen,kidney and skin were all significantly increased,indicating that these organs might play an essential role in response to high-temperature stress in A.ruthenus.

关 键 词：中华鲟(Acipenser sinensis) 热休克蛋白 HSP30 表达特征 温度胁迫 

分 类 号：S917.4[农业科学—水产科学]