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中国明对虾过氧化物酶 4 的分子特征及其对白斑综合征病毒感染的免疫反应和抗氧化活性。

Molecular characterization, immune response against white spot syndrome virus infection of peroxiredoxin 4 in Fenneropenaeus chinensis and its antioxidant activity.

机构信息

Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, 106 Nanjing Road, Qingdao 266071, China.

Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, 7 Nanhai Road, Qingdao 266071, China.

出版信息

Fish Shellfish Immunol. 2014 Mar;37(1):38-45. doi: 10.1016/j.fsi.2013.12.026. Epub 2014 Jan 10.

Abstract

Peroxiredoxins (Prx) are a family of antioxidant proteins and perform important functions in intracellular signal transduction. Here, we report a Prx gene from Chinese shrimp Fenneropenaeus chinensis. The full-length cDNA of FcPrx gene contained an open reading frame of 735 bp encoding a polypeptide of 275 amino acids. The molecular mass of the deduced amino acid of FcPrx is 27445.43 Da with an estimated pI of 5.71. Sequence comparison showed that the FcPrx shares high identities with Prx IVs and it was named FcPrx4. A real-Time PCR (qRT-PCR) assay was developed to assess the mRNA expression of FcPrx4 in different tissues and temporal expression in hemocytes and hepatopancreas of F. chinensis challenged by white spot syndrome virus (WSSV). Transcripts of FcPrx4 can be detected in all tissues examined. The expression of FcPrx4 showed significant up-regulation in shrimp hemocytes and hepatopancreas after artificial infection with WSSV. A fusion protein containing FcPrx4 was produced in vitro and was confirmed by Matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS) assay. And activity analysis indicated that the recombinant FcPrx4 proteins can reduce H2O2 in the presence of dithiothreitol.

摘要

过氧化物还原酶(Prx)是抗氧化蛋白家族,在细胞内信号转导中发挥重要功能。本研究报告了中国对虾(Fenneropenaeus chinensis)的一种 Prx 基因。FcPrx 基因的全长 cDNA 包含一个 735bp 的开放阅读框,编码 275 个氨基酸的多肽。FcPrx 推导的氨基酸的分子量为 27445.43Da,估计等电点为 5.71。序列比较表明,FcPrx 与 Prx IVs 具有高度同源性,因此将其命名为 FcPrx4。建立了实时 PCR(qRT-PCR)检测方法,评估了 FcPrx4 在不同组织中的 mRNA 表达水平,以及在中国对虾血细胞和肝胰腺中对白斑综合征病毒(WSSV)刺激的时间表达。在检测的所有组织中都可以检测到 FcPrx4 的转录本。人工感染 WSSV 后,FcPrx4 在对虾血细胞和肝胰腺中的表达显著上调。体外表达并经基质辅助激光解吸电离质谱(MALDI-MS)检测证实了含有 FcPrx4 的融合蛋白。活性分析表明,重组 FcPrx4 蛋白可以在二硫苏糖醇存在的情况下还原 H2O2。

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