| 摘要: |
| 铜锌超氧化物歧化酶（Superoxide dismutase，SOD）是一种广泛存在于生物体中最主要的抗氧化酶之一，在抵御过多活性氧簇对机体毒害的过程中起重要作用。本研究以太洋真宽水蚤为研究对象，采用RT-PCR与RACE方法克隆得到太平洋真宽水蚤Cu/Zn SOD基因全长cDNA序列。Cu/Zn SOD （GenBank登录号：MF289343）基因序列全长837bp，其中完全开放阅读框为456bp编码152个氨基酸，5'非编码区长297bp，3'非编码区长84bp，分子量约为15.050kDa，理论等电点为5.73。序列比较表明，太平洋真宽水蚤Cu/Zn SOD cDNA的氨基酸序列与其他甲壳动物的一致性较高；存在8个蛋白翻译后修饰位点及蛋白家族标签序列，无跨膜结构域与信号肽。在重金属铬与镍胁迫下，通过实时荧光定量PCR对太平洋真宽水蚤Cu/Zn SOD mRNA体内表达特点分析显示，其表达量均呈现先升后降趋势，在暴露12小时后表达量达到峰值；铬胁迫下的表达量略高于镍；联合胁迫下呈现出拮抗作用。本研究结果将有利于深入探讨桡足类Cu/Zn SOD基因的结构与功能，为进一步研究抗氧化分子机理奠定基础。 |
| 关键词: 铜锌超氧化物歧化酶 太平洋真宽水蚤 基因克隆 金属胁迫 实时荧光定量 |
| DOI：10.11693/hyhz20171000261 |
| 分类号:Q789;Q955 |
| 基金项目:国家海洋局公益性行业科研专项，201505025号；海洋科学浙江省重中之重学科开放课题资助，20130105号。 |
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| CLONING AND EXPRESSION ANALYSIS OF Cu/Zn SOD GENE OF EURYTEMORA PACIFICA UNDER METAL STRESS |
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HU Li-Teng, XIA Li-Ping, WU Min-Min, CHEN Jin, ZHANG Jian-She
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Marine Science College of Zhejiang Ocean University, National Engineering Research Center of Marine Facilities Aquaculture, Zhoushan 316022, China
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| Abstract: |
| Copper zinc superoxide dismutase (SOD) is one of the important antioxidant enzymes widely existing in organisms, and plays an important role in preventing excessive reactive oxygen species from poisoning the organism. In this study, Eurytemora pacifica was used as the research subject, and the full-length cDNA sequence of Cu/Zn SOD gene was obtained by RT-PCR and RACE methods. The full-sequence of Cu/Zn SOD (GenBank accession number:MF289343) was 837bp, the complete open reading frame of 456bp encoding 152 amino acids, 5'-3' non encoding region of 297bp encoding region of 84bp, the molecular weight of 15.050kDa and isoelectric point of 5.73. Sequence comparison indicated high consistency between the amino acid sequences of Cu/Zn SOD cDNA with other crustacean animal; there are 8 protein post-translational modification and protein family sequence tagged sites, no signal peptide and transmembrane domain. Cu/Zn SOD mRNA expression analysis of E. pacifica under chromium or nickel stress showed that the value of Cu/Zn SOD increased first and then decreased, and reached a peak in 12 hours. The expression in chromium stress is slightly higher than that in nickel stress. In addition, under nickel-chromium joint stress, it showed antagonistic effect. The results could be helpful to explore the structure and function of the copepod Cu/Zn SOD gene, and to offer theoretical basis for studying the molecular mechanism of antioxidant. |
| Key words: copper zinc superoxide dismutase Eurytemora pacifica gene cloning metal stress RT-PCR |
