Centre for Biotechnology in Agriculture Research, Division of Genetics & Molecular Biology, Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603 Kuala Lumpur, Malaysia.
Fish Shellfish Immunol. 2012 Jul;33(1):121-9. doi: 10.1016/j.fsi.2012.04.010. Epub 2012 Apr 28.
In this study, we have reported a full length of peroxiredoxin (designated MrPrdx) gene, identified from the transcriptome of freshwater prawn Macrobrachium rosenbergii. The complete gene sequence of the MrPrdx is 940 base pairs in length, and encodes 186 amino acids. MrPrdx contains a long thioredoxin domain in the amino acid sequence between 34 and 186. The gene expressions of MrPrdx in healthy and the infectious hypodermal and hematopoietic necrosis virus (IHHNV) challenged M. rosenbergii were examined using quantitative real time polymerase chain reaction. MrPrdx is highly expressed in all the other tissues of M. rosenbergii considered for analysis and the highest in gills. The expression is strongly up-regulated in gills after IHHNV infection. To understand MrPrdx functional properties, the recombinant MrPrdx protein was expressed in Escherichia coli BL21 (DE3) and purified. A peroxidise activity assay was conducted using recombinant MrPrdx protein at different concentrations. This peroxidises activity showed that the recombinant MrPrdx is a thiol-dependant protein. Additionally, this result showed that recombinant MrPrdx protein, as a secretory protein can remove H₂O₂ and protect DNA damage. This finding leads a possible way to propose the recombinant MrPrdx protein as an effective medicine for reactive oxygen species (ROS) related diseases.
在这项研究中，我们报道了一个全长的过氧化物酶（命名为 MrPrdx）基因，该基因是从淡水虾罗氏沼虾的转录组中鉴定出来的。MrPrdx 的完整基因序列长 940 个碱基对，编码 186 个氨基酸。MrPrdx 在氨基酸序列 34 到 186 之间包含一个长的硫氧还蛋白结构域。采用实时定量聚合酶链反应检测了健康罗氏沼虾和感染皮下及造血坏死病毒（IHHNV）的罗氏沼虾中 MrPrdx 的基因表达。MrPrdx 在分析中考虑的罗氏沼虾的所有其他组织中均高度表达，在鳃中表达最高。在 IHHNV 感染后，鳃中的表达强烈上调。为了了解 MrPrdx 的功能特性，将重组 MrPrdx 蛋白在大肠杆菌 BL21（DE3）中表达并纯化。在不同浓度的重组 MrPrdx 蛋白上进行了过氧化物酶活性测定。该过氧化物酶活性表明，重组 MrPrdx 是一种依赖巯基的蛋白。此外，该结果表明，重组 MrPrdx 蛋白作为一种分泌蛋白可以去除 H₂O₂并保护 DNA 免受损伤。这一发现为将重组 MrPrdx 蛋白作为与活性氧（ROS）相关疾病的有效药物提供了一种可能的途径。
