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

Journal of Tropical Oceanography ›› 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

• Marine Biology • Previous Articles

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

Abstract

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

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.

References

[1] .粱毅, 汪存信, 曲松生. 1995. 超氧化物歧化酶研究的新进展[J]．湖北化工, 3: 20-22.
[2] .田春美, 钟秋平. 2005. 超氧化物歧化酶的现状研究进展[J]. 中国热带医学, 5(8): 1730-1732.
[3] .吴军帅, 李培环, 李鼎立, 等. 2013. 桃果实铜锌超氧化物歧化酶基因PpCuZnSOD的克隆与分析[J]. 核农学报, 27(4): 0408-0417.
[4] .朱丹, 李宏俊, 高祥刚, 等. 2010. 文蛤胞内铜锌超氧化物歧化酶基因的克隆与序列分析[J]. 生物技术通报, 11: 123-133.
[5] .AINSWORTH T D, HOEGH-GULDBERG O, HERON S F, et al. 2008. Early cellular changes are indicators of pre-bleaching thermal stress in the coral host[J]. J Exp Mar Biol Ecol, 364(2): 63-71.
[6] .BROWN B E, DOWNS C A, DUNNE R P, et al. 2002. Exploring the basis of thermotolerance in the reef coral Goniastrea aspera [J]. Mar Ecol-Prog Ser, 242: 119-129.
[7] .CONNOLLY M A, CLAUSEN P A, LAZAR J G. 2006. Purification of RNA from animal cells using Trizol[J]. Cold Spring Harb Protoc. doi:10.1101/pdb. prot4114.
[8] .DASH B, METZ R, HUEBNER H J, et al. 2007. Molecular characterization of two superoxide dismutases from Hydra vulgaris [J]. Gene, 87(1-2): 93-108.
[9] .DOUGLAS A E. 2003. Coral bleaching—how and why?[J]. Mar Pollut Bull, 46(4): 385-392.
[10] .MERLE P L, SABOURAULT C, RICHIER S, et al. 2007. Catalase characterization and implication in bleaching of a symbiotic sea anemone[J]. Free Radical Biol Med, 42(2): 236-246.
[11] .PLANTIVAUX A, FURLA P, ZOCCOLA D, et al. 2004. Molecular characterization of two Cu/Zn-superoxide dismutases in a sea anemone[J]. Free Radical Biol Med, 2004, 37(8): 1170-1181.
[12] .RICHIER S, MERLE P L, FURLA P, et al. 2003. Characterization of superoxide dismutases in anoxia- and hyperoxia-tolerant symbiotic cnidarians[J]. Biochim Biophys Acta, 1621(1): 84-91.
[13] .SALIH A, LARKUM A, COX G, et al. 2000. Fluorescent pigments in corals are photoprotective[J]. Nature, 408: 850-853.
[14] .SHINZATO C, SHOGUCHI E, KAWASHIMA T, et al. 2011. Using the Acropora digitifera genome to understand coral responses to environmental change[J]. Nature, 2011, 476: 320-323.
[15] .SOREK M, YACOBI Y Z, ROOPIN M, et al. 2013. Photosynthetic circadian rhythmicity patterns of Symbiodium, the coral endosymbiotic algae[J]. Proc R Soc B, 280(1759): 1-9.
[16] .SOUTHWARD A J, TYLER P A, PUNG C M Y, et al. 2003. Advances in Marine Biology[M]. London: Academic Press: 184-212.
[17] .VENN A A, QUINN J, JONES R, et al. 2009. P-glycoprotein (multi- xenobiotic resistance) and heat shock protein gene expression in the reef coral Montastraea franksiin response to environmental toxicants[J]. Aquat Toxicol, 93(4): 188-195.
[18] .WEYDERT C J, WAUGH T A, RITCHIE J M, et al. 2006. Overexpression of manganese or copper-zinc superoxide dismutase inhibits breast cancer growth[J]. Free Radical Biol Med, 41(2): 226-237.
[19] .WILKINSON D G. 1999. In situ hybridization: A practical approach (Practical Approach Series)[M]. New York: Oxford University Press: 1-224.
[20] .YANG JIANGHUA, DONG SIMING, JIANG QICHEN, et al. 2013. Characterization and expression of cytoplasmic copper/zinc superoxide dismutase (CuZnSOD) gene under temperature and hydrogen peroxide (H 2 O 2 ) in rotifer Brachionus calyciflorus [J]. Gene, 518(2): 388-396.
[21] .ZELKO I N, MARIANI T J, FOLZ R J. 2002. Superoxide dismutase mutigene family: A comparison of the CuZn-SOD (SOD1), Mn-SOD (SOD2), and EC-SOD (SOD3) gene structures, evolution, and expression[J]. Free Radic Biol Med, 33(3): 337-349.

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

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