Centre for Biotechnology in Agriculture Research, Division of Genetics & Molecular Biology, Faculty of Science, University of Malaya, Kuala Lumpur, Malaysia.
Mol Biol Rep. 2012 Jun;39(6):6671-82. doi: 10.1007/s11033-012-1473-7.
In this study, we have reported a full length of small heat shock protein 37 (designated MrHSP37) gene, identified from the transcriptome database of freshwater prawn Macrobrachium rosenbergii. The complete gene sequence of the MrHSP37 is 2,425 base pairs in length, and encodes 338 amino acids. MrHSP37 contains a long heat shock protein family profile in the amino acid sequence between 205 and 288. The mRNA expressions of MrHSP37 in healthy and the infectious hypodermal and hematopoietic necrosis virus (IHHNV) challenged M. rosenbergii were examined using quantitative real time polymerase chain reaction (qRT-PCR). MrHSP37 is highly expressed in hepatopancreas and all the other tissues (walking leg, gills, muscle, stomach, haemocyte, intestine, pleopods, brain and eye stalk) of M. rosenbergii taken for analysis. The expression is strongly up-regulated after IHHNV challenge. To understand its biological activity, the recombinant MrHSP37 gene was constructed and expressed in Escherichia coli BL21 (DE3). The results of ATPase assay showed that the recombinant MrHSP37 protein exhibited apparent ATPase activity which increased with the concentration of the protein. And also the purified recombinant MrHSP37 protein was used for thermal aggregation assay (chaperone activity). It showed that the recombinant MrHSP37 protein is an active chaperone in this assay. Taken together, these results suggest that MrHSP37 is potentially involved in the immune responses against IHHNV challenge in M. rosenbergii.
在本研究中，我们从淡水虾Macrobrachium rosenbergii的转录组数据库中报告了全长的小分子热休克蛋白 37（命名为 MrHSP37）基因。MrHSP37 的完整基因序列长 2425 个碱基对，编码 338 个氨基酸。MrHSP37 在氨基酸序列 205 到 288 之间含有一个长的热休克蛋白家族特征。使用定量实时聚合酶链反应（qRT-PCR）检测了健康和感染皮下及造血坏死病毒（IHHNV）的罗氏沼虾 MrHSP37 的 mRNA 表达。MrHSP37 在罗氏沼虾的所有组织（步行足、鳃、肌肉、胃、血细胞、肠、尾扇、脑和眼柄）中高度表达，用于分析。在 IHHNV 攻击后，表达水平强烈上调。为了了解其生物学活性，构建了重组 MrHSP37 基因并在大肠杆菌 BL21（DE3）中表达。ATPase 测定结果表明，重组 MrHSP37 蛋白表现出明显的 ATPase 活性，且随着蛋白浓度的增加而增加。此外，还对纯化的重组 MrHSP37 蛋白进行了热聚集测定（伴侣活性）。结果表明，在该测定中，重组 MrHSP37 蛋白是一种具有活性的伴侣。综上所述，这些结果表明 MrHSP37 可能参与了罗氏沼虾对 IHHNV 攻击的免疫反应。
