草鱼Wap65基因的克隆和鉴定  

Molecular Cloning and Identification of the Wap65 Gene from Grass Carp(Ctenopharyngodon idella)

作　　者：赵风云[1] 吴初新[1] 朱玉娇[1] 胡成钰[1] 

机构地区：[1]南昌大学生命科学与食品工程学院,江西南昌330031

出　　处：《水产科学》2011年第5期281-286,共6页Fisheries Science

基　　金：江西省重点科技攻关项目(20061B0260301);江西省创新性试验专项资金资助项目

摘　　要：经Poly I∶C诱导的草鱼肝肾cDNA文库中筛选并克隆了一个Wap65(HM629798)。草鱼Wap65全长cDNA为1510 bp,包括5’非编码区55 bp,3’非翻译区327 bp,最大开放阅读框为1128bp。草鱼Wap65是一个376 aa的蛋白质,其中,第56～95 aa、102～153 aa、155～199 aa、200～243 aa、262～305 aa和307～352 aa为重复血红素结合域。进化树显示,草鱼Wap65与斑点叉尾鱼回Wap65-2进化关系最近。定量PCR研究草鱼Wap65的表达特征,Wap65在肝中有较强的本底表达,而在脾、肾、脑和肠等组织中该基因表达非常微弱。34℃热激2 h后,在脾脏中,Wap65的表达量有一定的上调;在肝脏中的该基因的相对表达量大量增加,而在其他受检组织中Wap65的表达无明显变化。因此,草鱼Wap65主要在肝脏中表达。以上结果表明,本研究克隆的草鱼Wap65可能是Wap65-2。The warm temperature acclimation related 65 ku protein（Wap65）is a stress protein involved in thermoregulation and immune response and plays a crucial role in heat and cold adaption in teleost. In this report,we cloned and identified a Wap65 （CiWap65, HM629798）in cDNA library（induced by Poly I ： C） constructed from liver and kidney tissues of grass carp（Ctenopharyngodon idella）. The full-length cDNA of Wap65 is 1510 bp with 55 bp in the 5＇ UTR and 327 bp in the 3＇UTR. The dedued ORF is 1128 bp encoding a protein of 376 aa which contains six repeat heine binding domains at the position of 56-95 aa,102- 153 aa, 155 - 199 aa, 200 ~ 243 aa, 262 - 305 aa and 307 - 352 aa, respectively. The phylogenetic analysis showed that CiWap65 grouped tightly with catfish Wap65-2 in the same branch. The qPCR suggested that the CiWap65 had strong basal expression in liver, but very weak in spleen, kidney, brain and intestine. By heat shocking at 34 ℃ for 2 hours,the expression slightly increased in spleen and was significantly up-regulated in liver but no obvious changs in other tissues tested. So CiWap65 was mainly expressed in liver. These datas support the idea that the CiWap65 isolated in this study probably is Wap65-2.

关 键 词：Wap65 应激蛋白 血红素结合蛋白 草鱼 

分 类 号：Q785[生物学—分子生物学]