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dc.contributor.author于坤
dc.contributor.author叶海辉
dc.contributor.author巩杰
dc.contributor.author黄辉洋
dc.contributor.author林思恒
dc.date.accessioned2016-05-17T02:28:50Z
dc.date.available2016-05-17T02:28:50Z
dc.date.issued2014-5-28
dc.identifier.citation厦门大学学报(自然科学版),2014,(3):146-152
dc.identifier.issn0438-0479
dc.identifier.otherXDZK201403028
dc.identifier.urihttps://dspace.xmu.edu.cn/handle/2288/104751
dc.description.abstract腺苷酸转移酶(AnT)是线粒体中最丰富的蛋白质之一,其主要作用是介导AdP/ATP在细胞质和线粒体基质之间的运输.为探讨该基因在拟穴青蟹(SCyllA PArAMAMOSAIn)低温适应中的作用,采用反转录PCr(rT-PCr)、CdnA末端快速扩增技术(rACE)等技术,从拟穴青蟹中获得了AnT2的CdnA全长序列,并运用实时荧光定量PCr(QPCr)技术检测了AnT2在不同组织和不同温度下的表达谱.该序列全长1 348bP,开放阅读框(Orf)为930bP,编码309个氨基酸残基.同源分析显示,该蛋白具有3个保守的线粒体跨膜功能结构域,形成能量分子传导转运通道,催化细胞质中AdP和线粒体内ATP的跨膜交换.QPCr结果表明AnT2基因在拟穴青蟹多个组织中均有表达,且表达量不同,表明AnT2基因具有组织表达特异性.第1期仔蟹在10,15,20,25℃不同温度条件下,在1,3,12,24,36H,10和15℃组表达量显著低于20和25℃组(P<0.05);且在1,3,6,12,24H,10℃组的表达量显著低于15℃组表达量(P<0.05).15℃组在48H内,AnT2呈现高低起伏的表达模式.拟穴青蟹第1期仔蟹AnT2基因表达变化,提示该基因与能量代谢功能相关,可能参与拟穴青蟹低温胁迫应答.
dc.description.abstractAdenine nucleotide translocase(ANT)is one of the most abundant proteins in mitochondria,and its role is largely to mediate the ADP/ATP transportation between the cytoplasm and the mitochondrial matrix.To determine the role of ANT2 gene under low temperature adaptation in the mud crab,Scylla paramamosain,ANT2 gene was firstly cloned by RT-PCR and RACE,and its expression profiles were analyzed by real time quantitative(qPCR).Results showed that the full length ANT2 cDNA was 1 348bp with 930bp open reading frame encoding 309 amino acids.Homologous analysis showed that ANT2 protein had three conserved mitochondria-type cross-membrane structure domains,which formed the energy transmission channel for the exchanges of ADP/ATP between the cytoplasm and the mitochondria across the membrane.qPCR analysis indicated that ANT2 was expressed in various tissues of the S.paramamosain with tissue specificities.Moreover,the expression of ANT2 was measured at various time points when S.paramamosainjuveniles were incubated at different low temperatures(10,15,20,and 25℃).The results showed that the expression of ANT2 at 10 and 15℃treatments was significantly lower than at 20 and 25℃ treatments at 1,3,12,24,and 36h(p<0.05).At 10℃treatment,the expression of ANT2 was significantly lower than at 15℃ treatment at 1,3,6,12,and 24h(p<0.05).At 15℃ treatment,the expression of ANT2 was fluctuant within 48h.The expression pattern of ANT2 under low temperatures indicated that it might be associated with energy metabolism and participate in stress response of low temperature in S.paramamosain juveniles.
dc.description.sponsorship福建省自然科学基金(2012J01147); 福建省科技计划重点项目(2008N0040)
dc.language.isozh_CN
dc.subject拟穴青蟹
dc.subjectANT2
dc.subject低温胁迫
dc.subject实时荧光定量PCR(qPCR)
dc.subjectScylla paramamosain
dc.subjectANT2
dc.subjectlow-temperature stress
dc.subjectreal time quantitative(qPCR)
dc.title拟穴青蟹ANT2基因的克隆与低温胁迫表达分析
dc.title.alternativeCloning of Adenine Nucleotide Translocase 2 (ANT2) and Its Expression Under Low Temperature Stress in the Mud Crab,Scylla paramamosain
dc.typeArticle


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