背角无齿蚌铁蛋白基因的克隆及Fe3+和Cd2+对其表达的影响
Cloning Characterizations of Ferritin Gene from Freshwater Clam Anodonta woodiana and Effect of Fe3+ and Cd2+ on Expressions of Ferritin
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摘要: 铁蛋白在参与铁稳态调控、细胞增殖、蛋白质翻译和免疫应答过程中发挥积极作用。为了探讨Fe3+和Cd2+对背角无齿蚌(Anodonta woodiana)的分子毒理效应，克隆出AwFer1和AwFer2全基因序列，分析Fe3+和Cd2+对AwFer1和AwFer2表达的影响。结果显示，在本研究中AwFer1包含780个碱基的核苷酸序列，174个氨基酸编码的开放阅读框；AwFer2包含874个碱基的核苷酸序列，174个氨基酸组成的开放阅读框。AwFer1和AwFer2具有高度保守的铁反应元件序列，5’非编码区共有序列上游存在一个CAGUGA茎环结构和2个凸起点，蛋白序列包括一个铁结合区域标签(59EEREHAEKFMKYQNKRGGR77)。与对照组相比，Fe3+和Cd2+处理组动物肝胰脏和鳃中AwFer1和AwFer2表达水平显著上调。以上结果表明，Fe3+和Cd2+处理对背角无齿蚌AwFer1和AwFer2表达水平具有显著的诱导作用，其原因与维持机体内铁的稳态效应和免疫防御作用有关。Abstract: Ferritin exerts positive effects in the regulation of iron homeostasis, cell proliferation, protein translation and immune response. AwFer1 and AwFer2, two ferritin subunit genes, were isolated from freshwater bivalve Anodonta woodiana. In this study, to explore the molecular toxicological effects for Fe3+ and Cd2+ on Anodonta woodiana, the whole genomic sequences of AwFer1 and AwFer2 were cloned, and the effects of Fe3+ and Cd2+ on the expressions of AwFer1 and AwFer2 proteins were analyzed. The results showed that the nucleotide sequences of AwFer1 and AwFer2 were respectively containing 780 bp and 874 bp. and the open reading frames (ORF) were coded by 174 amino acids. The highly conserved iron response element sequences, a CAGUGA stem-loop sequence and two bulges located upstream of the consensus sequence in the 5’ UTR and a putative iron binding region signature (59EEREHAEKFMKYQNKRGGR77), existed in the whole genomic sequences of AwFer1 and AwFer2. Compared with the control group, the expressions of AwFer1 and AwFer2 proteins extracted from the hepatopancreas and gill were significantly increased in the Fe3+and Cd2+ treated group. The results suggested that the induction effects of the expressions of AwFer1 and AwFer2 proteins were exerted by Fe3+and Cd2+, which are associated with the maintaining iron homeostasis and enhancing immune defense in our internal environment.
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Key words:
- Anodonta woodiana /
- AwFer1 /
- AwFer2 /
- iron homeostasis /
- immune defense
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