从日本囊对虾中克隆和鉴定了一种胞浆锰超氧化物歧化酶（cytMnSOD）和线粒体锰超氧化物歧化酶（mtMnSOD）。
Molecular cloning and characterization of a cytosolic manganese superoxide dismutase (cytMnSOD) and mitochondrial manganese superoxide dismutase (mtMnSOD) from the kuruma shrimp Marsupenaeus japonicus.
机构信息
Department of Aquaculture, College of Life Sciences, National Taiwan Ocean University, Keelung 202, Taiwan, ROC.
出版信息
Fish Shellfish Immunol. 2010 Jan;28(1):143-50. doi: 10.1016/j.fsi.2009.10.012. Epub 2009 Oct 14.
A cytosolic manganese superoxide dismutase (cytMnSOD) gene and a mitochondrial manganese superoxide dismutase (mtMnSOD) gene were cloned from the kuruma shrimp Marsupenaeus japonicus using reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) method. The open reading frame (ORF) of cytMnSOD is 861 bp and encodes a 287 amino acids (aa) protein with a 61 aa leader sequence, whereas the ORF of mtMnSOD is 663 bp and encodes a 221 aa protein with a 21 aa mitochondrial-targeting sequence in the N-terminus. The calculated molecular mass of translated protein of cytMnSOD and mtMnSOD is 31.4 kDa and 24.3 kDa with an estimated pI of 5.62 and 7.27, respectively. The deduced amino acid sequence of cytMnSOD has similarity of 50.2% to that of mtMnSOD. Both cytMnSOD and mtMnSOD contain a manganese superoxide dismutase domain (DVWEHAYY), and four conserved amino acids responsible for binding manganese. Both cytMnSOD and mtMnSOD of M. japonicus were expressed in haemocytes, eyestalk, muscle, intestine, gill, and hepatopancreas. Both cytMnSOD and mtMnSOD transcripts in haemocytes of M. japonicus significantly increased 6 h after injection of Vibrio alginolyticus, and 12 h after injection of beta-glucan, indicating induction of SOD system response in a short time.
利用反转录-聚合酶链反应（RT-PCR）和快速扩增 cDNA 末端（RACE）方法，从日本对虾（Marsupenaeus japonicus）中克隆出细胞质锰超氧化物歧化酶（cytMnSOD）基因和线粒体锰超氧化物歧化酶（mtMnSOD）基因。cytMnSOD 的开放阅读框（ORF）为 861 bp，编码一个 287 个氨基酸（aa）的蛋白质，带有 61 aa 的前导序列，而 mtMnSOD 的 ORF 为 663 bp，编码一个 221 个氨基酸的蛋白质，在 N 端带有 21 aa 的线粒体靶向序列。cytMnSOD 和 mtMnSOD 翻译蛋白的计算分子量分别为 31.4 kDa 和 24.3 kDa，估计 pI 分别为 5.62 和 7.27。cytMnSOD 的推导氨基酸序列与 mtMnSOD 的相似性为 50.2%。cytMnSOD 和 mtMnSOD 均含有锰超氧化物歧化酶结构域（DVWEHAYY）和四个负责结合锰的保守氨基酸。日本对虾的 cytMnSOD 和 mtMnSOD 均在血细胞、眼柄、肌肉、肠、鳃和肝胰腺中表达。日本对虾血细胞中 cytMnSOD 和 mtMnSOD 的转录本在注射溶藻弧菌 6 小时后和注射β-葡聚糖 12 小时后显著增加，表明 SOD 系统在短时间内被诱导。
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