凡纳滨对虾ATF-2分子克隆及表达特征分析  

Molecular cloning and expression analysis of ATF-2 from the shrimp Litopenaeus vannamei

作　　者：姚德福 阮灵伟[2] 施泓[2] 

机构地区：[1]厦门大学生命科学学院,福建厦门361005 [2]国家海洋局第三海洋研究所国家海洋局海洋生物遗传资源重点实验室,福建厦门361005

出　　处：《应用海洋学学报》2016年第2期190-197,共8页Journal of Applied Oceanography

基　　金：现代农业产业技术体系专项资金资助项目(CARS-47);国家海洋局第三海洋研究所基本科研业务费专项资金资助项目(2014020)

摘　　要：激活转录因子-2(ATF-2)作为细胞内应激活化蛋白激酶下游效应分子,参与调控了生物体诸如细胞增殖、凋亡以及免疫炎症反应等许多细胞生物学过程.本研究首次从凡纳滨对虾(Litopenaeus vannamei)中克隆得到了ATF-2基因全长编码序列,命名为Lv ATF-2.序列分析表明,Lv ATF-2基因开放阅读框长1 719 bp,可编码572个氨基酸.其编码的蛋白具有特征性的锌指结构和碱性亮氨酸拉链结构域,并且在N端具有两个保守的苏氨酸磷酸化位点(Thr71和Thr73).进一步的系统进化树分析显示,Lv ATF-2蛋白与脊椎动物及软体动物ATF-2蛋白聚为一簇,且与软体动物ATF-2蛋白亲缘关系更近.半定量RT-PCR结果显示,Lv ATF-2基因在所检测的各个组织中均有转录,但在鳃和肠道中转录量较高.此外Lv ATF-2基因在白斑综合症病毒(WSSV)感染早期转录水平显著下降,提示该基因与WSSV感染密切相关,可能参与了对虾应答病毒的免疫反应过程.本文通对Lv ATF-2基因的克隆与表达特征分析,为将来研究该基因在对虾免疫应答过程中的功能提供了依据.ATF-2,as a downstream effector of cellular stress-activated protein kinases,participates in the regulation of numerous cell processes,including cell proliferation and differentiation,cell cycle,apoptosis and immune inflammatory response. In the present study,a ATF-2 gene from Litopenaeus vannmei was cloned for the first time and designated as Lv ATF-2. Sequence analysis showed that the open reading frame of Lv ATF-2 gene was 1 719 bp in length and encoded a protein with 572 amino acids. The Lv ATF-2 protein contained characteristic zinc finger and b ZIP domains,and two conserved threonine phosphorylation sites（ Thr71 and Thr73） at the N-terminus. Phylogenetic tree analysis further displayed that Lv ATF-2 protein was clustered with vertebrate and mollusc ATF-2 protein and showed a closer relationship with ATF-2 protein of the mollusc. Semi-quantitative RT-PCR showed that Lv ATF-2 mRNAs were transcribed in all examined tissues,but the mRNA level of Lv ATF-2 gene was relatively higher in intestine and gill tissues. In addition,the transcriptional level of Lv ATF-2 gene was remarkably down-regulated during the early stage of WSSV infection,suggesting that Lv ATF-2 gene was closely related to WSSV infection and might be involved in immune response upon viral infection. Our study on Lv ATF-2 gene might have provided useful clue for exploring its function in shrimp immune response in future.

关 键 词：海洋生物学 凡纳滨对虾 激活转录因子-2(ATF-2) 白斑综合症病毒(WSSV) 分子克隆 

分 类 号：P735[天文地球—海洋生物学]