从本研究前期构建的短蛸(Octopusocellatus)cDNA文库中克隆得到过氧化物还原酶(OoPrx-4)基因的cDNA全长,该基因cDNA全长934bp,包括5’非编码区(UTR)19bp,3'UTR177bp,开放阅读框(ORF)738bp,共编码245个氨基酸,理论等电点为6.65,预测分子量27.1kDa。采用实时荧光定量PCR法分析了OoPrx-4基因在短蛸各组织及鳗弧菌胁迫下的表达规律,结果表明,OoPrx-4在短蛸血细胞、肌肉、系统心脏、鳃、胃、肾囊、性腺、外套膜和肝胰腺等检测组织中都有表达,其中在肝胰腺的表达量最高。经鳗弧菌刺激后,Prx-4在血细胞中的表达量分别在6h和48h出现了两次明显上调。Prx-4作为抗氧化酶可能在减少机体因抵御鳗弧菌胁迫所产生的过氧化物方面发挥重要的作用。
The full-length cDNA encoding peroxiredoxin-4 (designated as OoPrx-4) was cloned from Octopus ocella- tus. The full-length cDNA was 934bp, containing a 19bp 5' untranslated region (UTR), a 177 bp 3UTR with a poly (A) tail, and a 738 bp open reading frame (ORF) of encoding a polypeptide of 245 amino acids. The predicted molecular mass of the amino acid is 27. lkDa with an estimated pI of 6.65. The expression patterns of OoPrx-4, in both normal and Listonella anguillarum-challenged tissues were then characterized by Real-Time PCR (RT-PCR). OoPrx-4 was found expressed in many tissues, including hemocyte, muscle, systemic heart, gill, stomach, saccus renalis, gonad, mantle, and hepatopancreas, and especially, it was highly expressed in hepatopancreas. The mRNA expression of OoPrx-4 in hemocytes was signifi- cantly up-regulated (P〈0.01) at 6h and 48h post challenge. The results indicate that OoPrx-4 played an important role in scavenging ROS generated during L. anguillarum invasion.