Cloning and identification of four Mu-type glutathione S-transferases from the giant freshwater prawn Macrobrachium rosenbergii
Cloning and identification of four Mu-type glutathione S-transferases from the giant freshwater prawn Macrobrachium rosenbergii
复制标题
罗氏沼虾四种Mu型谷胱甘肽S-转移酶的克隆与鉴定
DOI：
10.1016/j.fsi.2013.05.012
复制
发表时间：
2013-08-01
影响因子：
4.7
通讯作者：
Ren, Qian
中科院分区：
文献类型：
--
作者：
Hui, Kai-Min;Hao, Fang-Yuan;Ren, Qian
谷胱甘肽s -转移酶（GSTs）是细胞解毒系统的重要组成部分，因为它们具有保护生物体免受活性氧（ROSs）毒性的能力。从罗氏沼虾肝胰腺中克隆出4个与mu型GSTs相似的GSTs （MrMuGST1-MrMuGST4）。这四种gst的氨基酸长度分别为219、216、218和219个。MrMuGST1-MrMuGST4蛋白都在n端有一个g位点，在c端有一个h位点。系统发育分析表明，4个mu型gst可分为两个不同的支系（MrMuGST2为一个支系，MrMuGST1、MrMuGST3和MrMuGST4为另一个支系）。尽管如此，没有检测到正选择位点，但在少数MuGST基因中发现了快速进化。逆转录聚合酶链反应（RT-PCR）结果显示，MrMuGST1和MrMuGST2转录本在所有检测组织中均有表达，而MrMuGST3和MrMuGST4仅主要在肝胰腺和肠道中表达。定量RT-PCR分析显示，MrMuGST1和MrMuGST2在鳗弧菌攻毒后下调，而MrMuGST3和MrMuGST4在攻毒2 h后迅速上调。我们的结果表明，不同的mu型gst可能以不同的方式响应弧菌的挑战。(C) 2013 Elsevier Ltd.版权所有。
Glutathione S-transferases (GSTs) are essential components of the cellular detoxification system because of their capability to protect organisms against the toxicity of reactive oxygen species (ROSs). Four different GSTs (MrMuGST1-MrMuGST4) showing similarities with Mu-type GSTs were cloned from the hepatopancreas of Macrobrachium rosenbergii. These four GSTs have 219, 216, 218 and 219 amino acids in length, respectively. MrMuGST1-MrMuGST4 proteins all have a G-site in the N-terminus and an H-site in the C-terminus. Phylogenetic analysis reveals that four Mu-type GSTs are classified into two different clades (MrMuGST2 one clade; MrMuGST1, MrMuGST3 and MrMuGST4 other clades). Nonetheless, no site under positive selection was detected but rapid evolution was found in the few of MuGST genes. Reverse transcription-polymerase chain reaction (RT-PCR) results showed that MrMuGST1 and MrMuGST2 transcripts were expressed in all detected tissues, however, MrMuGST3 and MrMuGST4 were just mainly expressed in hepatopancreas and intestines. Quantitative RT-PCR analysis showed that MrMuGST1 and MrMuGST2 were down-regulated upon Vibrio anguillarum challenge, whereas MrMuGST3 and MrMuGST4 were quickly up-regulated 2 h after the Vibrio challenge. Our results imply that different Mu-type GSTs may respond to Vibrio challenge with different manners. (C) 2013 Elsevier Ltd. All rights reserved.