斑节对虾Pellino基因的克隆及其在不同胁迫条件下的表达分析

Molecular cloning and expression analysis of Pellino in black tiger shrimp(Penaeus monodon) under different stress

Pellino蛋白家族,是一类高度保守的E3泛素连接酶,在泛素化和先天性免疫中发挥重要作用。该研究通过RACE技术得到斑节对虾(Penaeus monodon)泛素连接酶Pellino基因的c DNA全长。该基因序列全长1 961 bp,编码区序列长1 299 bp,编码432个氨基酸,5’非编码区(UTR)为89 bp,3’UTR为573 bp。通过qRT-PCR技术,研究了PmPellino基因在斑节对虾不同组织中的表达水平,并研究了其在不同浓度氨氮胁迫下和不同微生物刺激下的表达情况。结果显示,PmPellino基因在各组织中均有表达,在鳃组织中表达量最高。急性氮氮胁迫后,PmPellino在肝胰腺中的表达量显著上调(P<0.01),但在鳃中的表达被抑制(P<0.01)。哈维氏弧菌(Vibrio harveyi)可显著激活PmPellino在鳃中的表达,抑制其在肝胰腺中的表达。鳗弧菌(V. anguillarum)可显著抑制PmPellino在肝胰腺中的表达,而对PmPellino在鳃中的表达无显著影响。金黄色葡萄球菌(Staphylococcus aureus)可以显著激活PmPellino在肝胰腺和鳃中的表达。结果表明PmPellino可以激活免疫应答通路,在免疫防御中发挥重要作用。

Pellino protein is a member of highly conserved E3 ubiquitin ligase which plays an important role in ubiquitination and innate immunity. In this study, we cloned and identi?ed a crustacean Pellino from Penaeus monodon(PmPellino). The full-length cDNA of PmPellino consisted of 1 961 bp with an 89 bp 5’UTR, a 573 bp 3’UTR and a 1 299 bp open reading frame encoding 432 amino acids. PmPellino mRNA was detected in all the tissues examined by real time PCR(highest in gill). The expression of PmPellino in hepatopancreas was significantly up-regulated after acute ammonia nitrogen stress(P<0.01), but was inhibited in gill(P<0.01). Vibrio harveyi could significantly activate the expression of PmPellino in gill and inhibit its expression in hepatopancreas.V. anguillarum could significantly inhibit the expression of PmPellino in hepatopancreas, but there was no sigficant difference in gill with the control group. Staphylococcus aureus could significantly activate the expression of PmPellino in both hepatopancreas and gill. The results indicate that PmPellino might play an important role in the immunity of P. monodon.