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蚯蚓(参环毛蚓,E. 佩里埃)镉诱导型金属硫蛋白-2基因的克隆、表征及表达

Cloning, characterization, and expression of cadmium-induced metallothionein-2 gene from earthworm Pheretima aspergillum (E. Perrier).

作者信息

Gong L, Li W, Li J, Li W E, Wu W R, Yu L W

机构信息

School of Chinese Materia Medica, Guangzhou University of Chinese Medicine, Guangzhou, China.

出版信息

Genet Mol Res. 2015 Dec 14;14(4):16782-92. doi: 10.4238/2015.December.14.5.

Abstract

Metallothioneins (MTs) are ubiquitous metal-binding, cysteine-rich proteins, associated with metal accumulation and thus providing protection against toxic heavy metals such as cadmium (Cd). To investigate the mechanisms of enrichment of Cd in the earthworm Pheretima aspergillum, we isolated and cloned metallothionein-2 (MT-2) cDNA (538 bp) from P. aspergillum, analyzed its sequence, and examined MT-2 transcription levels by relative quantitative real-time PCR under different concentrations of Cd. The sequence of P. aspergillum MT-2 cDNA and its putative amino acid sequence were highly similar to sequences from other earthworms. The induction with Cd increased the MT-2 gene transcription level in a dose-dependent manner. In addition, earthworm recombinant MT-2 exhibited high Cd bioaccumulation ability in vitro. These results suggested that MT-2 plays an important role in tolerance and accumulation of Cd in P. aspergillum.

摘要

金属硫蛋白(MTs)是普遍存在的、富含半胱氨酸的金属结合蛋白,与金属积累相关,从而提供对镉(Cd)等有毒重金属的保护作用。为了研究蚯蚓(参环毛蚓)中镉的富集机制,我们从参环毛蚓中分离并克隆了金属硫蛋白-2(MT-2)的cDNA(538 bp),分析了其序列,并通过相对定量实时PCR检测了不同镉浓度下MT-2的转录水平。参环毛蚓MT-2 cDNA序列及其推测的氨基酸序列与其他蚯蚓的序列高度相似。镉诱导以剂量依赖的方式增加了MT-2基因的转录水平。此外,蚯蚓重组MT-2在体外表现出高镉生物积累能力。这些结果表明,MT-2在参环毛蚓对镉的耐受性和积累中起重要作用。

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