日本对虾Fos基因的克隆及表达分析

Cloning and Expression Analysis of Fos in Kuruma Shrimp(Penaeus japonicus)

Fos是催化蛋白1家族中的一员,在动物免疫过程中起重要作用。本研究获得了日本对虾Fos的cDNA序列(命名为PjFos),其长1955 bp,其中5'非翻译区(UTR)长95 bp,3'UTR长348 bp,开放阅读框(ORF)长1512 bp,编码503个氨基酸。序列分析显示PjFos蛋白含有1个碱性亮氨酸拉链结构域(bZIP)和两个核定位信号,无跨膜结构和信号肽。蛋白高级结构预测Fos蛋白bZIP结构域的三级结构为α螺旋。多重序列比对结果表明PjFos与南美白对虾的Fos相似性最高(94%)。Fos的碱性亮氨酸拉链结构域在各物种中非常保守。系统进化树表明日本对虾与南美白对虾聚在一起。PjFos mRNA在各组织中都检测到表达量,表达量最高的组织为神经节,其次为肝胰腺和鳃。在哈维弧菌刺激下,PjFos在鳃中的表达量与对照组相比呈极显著上调(P<0.01),在第24小时PjFos的表达量达到最大值。这些结果表明PjFos可能参与了日本对虾的先天免疫。

Fos is one of member in activator protein 1(AP-1)family and played crucial functions in animals immune system.In this study,the c DNA sequence of Penaeus japonicus Fos gene(named PjFos)was obtained and analyzed.The full length c DNA was 1955 bp,including 5’untranslate region 95 bp,3 untranslate region 348 bp,open reading frame(ORF)1512 bp which encoding 503 amino acid.Sequence analysis revealed that PjFos protein contained a basic leucine zipper(bZIP)domain and two nuclear localization signal(NLS)sequences.No transmembrane domain and signal peptide were predicted successfully.The result of protein advanced structure prediction showed that bZIP domain was α helix.Multiple sequence alignment analysis showed that PjFos had the highest homology with Fos of Penaeus vannamei(94%)and bZIP domain was conserved among most species.Phylogenetic tree analysis discovered that P.japonicus Fos was close to P.vanamei.PjFos was ubiquitously expressed in all tested tissues.The highest expression level was detected in the ganglion,followed by hepatopancreas and gill.Under the challenge of Vibrio harveyi,the expression of PjFos in gill was significantly up-regulated compared with the control group(P<0.01),reached the maximum at 24 h.These results indicated that PjFos may play a key role in innate immunity of P.japonicus.