稀杯盔形珊瑚铜锌超氧化物歧化酶基因全长cDNA 序列的克隆与分析*

doi:10.11978/j.issn.1009-5470.2015.01.012

热带海洋学报 ›› 2015, Vol. 34 ›› Issue (1): 83-89.doi: 10.11978/j.issn.1009-5470.2015.01.012CSTR: 32234.14.j.issn.1009-5470.2015.01.012

• 海洋生物学 • 上一篇

稀杯盔形珊瑚铜锌超氧化物歧化酶基因全长cDNA 序列的克隆与分析^*

范程辉, 刘丽, 沈城, 郭昱嵩

广东海洋大学水产学院南海水产经济动物增养殖广东省普通高校重点实验室广东湛江 524088

收稿日期:2014-03-24 修回日期:2014-06-14 出版日期:2015-02-10 发布日期:2015-02-10
作者简介:范程辉(1987~), 男, 湖南省宜章县人, 硕士研究生, 主要从事海洋经济动物发育生物学的研究。E-mail: fchairain@126.com
基金资助:
国家海洋公益性行业科研专项(201105012); 广东省自然科学基金项目(S2011010000269); 广东省海洋渔业科技推广专项(A201308E02); 中央分成海域使用金支出项目201122093); 感谢广东省徐闻县徐闻珊瑚礁国家级自然保护区给予的帮助和支持

Molecular cloning and sequence analysis of the copper-zinc superoxide dismutase gene in Galaxea astreata

FAN Cheng-hui, LIU Li, SHEN Cheng, GUO Yu-song

Fisheries College, Guangdong Ocean University, Key Laboratory of Aquaculture in South China Sea for Aquatic Economic Animal of Guangdong Higher Education Institutes, Zhanjiang 524088, China

Received:2014-03-24 Revised:2014-06-14 Online:2015-02-10 Published:2015-02-10
Contact: 刘丽。E-mail: zjouliuli@163.com

摘要/Abstract

摘要： 利用反转录PCR和RACE法获得稀杯盔形珊瑚 Galaxea astreata的铜锌超氧化物歧化酶(copper-zinc superoxide dismutase, CuZnSOD)基因 cDNA全序列, 大小为730 bp。5°非编译区(5°-UTR)为111 bp, 3°-UTR为188 bp, 开放阅读框(open reading frame, ORF)长度为471bp, 编码156个氨基酸。编码的目的氨基酸序列中含有两个CuZnSOD蛋白家族信号, Cu²⁺分别与His49、His51、His66和His123配位, Zn²⁺则与His66、His74、His83和Asp86配位, Cys60和Cys149之间形成唯一的链内二硫键。SignalP 3.0 Server预测并未发现信号肽, TMHMM Server v. 2.0预测不存在跨膜结构, PSORT II Prediction预测位于细胞质的可能性为56.5%, 推测可能是胞内CuZnSOD。本研究结果为进一步揭示造礁石珊瑚的抗氧化分子机制奠定了基础。

关键词: 铜锌超氧化物歧化酶, 基因克隆, 序列分析, 稀杯盔形珊瑚Galaxea astreata

Abstract: The full-length cDNA sequence of copper-zinc superoxide dismutase (CuZnSOD) gene was obtained from Galaxea astreata using RT-PCR and RACE methods. The results revealed that the full-length cDNA of CuZnSOD gene was 730 bp including 111 bp 5°-UTR, 188 bp 3°-UTR and 471 bp open reading frame (ORF) encoding 156 amino acids. The deduced amino acid sequence contained two CuZnSOD protein family signals. Cu²⁺ integrated with His49, His51, His66, and His123, and Zn²⁺integrated with His66, His74, His83, and Asp86. The only intrachain disulfide bond was formed between Cys60 and Cys149. The signal peptides were not predicted by SignalP 3.0 Server, the transmembrane domains were not predicted by TMHMM Server v. 2.0, and the possibility of 56.5% was located in the cytoplasm by means of PSORT II Prediction, indicating that the deduced protein was intracellular CuZnSOD. The results will help to further reveal antioxidant molecular mechanism in Sceratinian corals.

Key words: CuZnSOD, gene cloning, sequence analysis, Galaxea astreata

范程辉, 刘丽, 沈城, 郭昱嵩. 稀杯盔形珊瑚铜锌超氧化物歧化酶基因全长cDNA 序列的克隆与分析^*[J]. 热带海洋学报, 2015, 34(1): 83-89.

FAN Cheng-hui, LIU Li, SHEN Cheng, GUO Yu-song. Molecular cloning and sequence analysis of the copper-zinc superoxide dismutase gene in Galaxea astreata[J]. Journal of Tropical Oceanography, 2015, 34(1): 83-89.

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稀杯盔形珊瑚铜锌超氧化物歧化酶基因全长cDNA 序列的克隆与分析^*

Molecular cloning and sequence analysis of the copper-zinc superoxide dismutase gene in Galaxea astreata

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