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巴西橡胶树金属硫蛋白基因HbMT2a的克隆与特性分析:其对非生物胁迫和重金属有不同响应

Cloning and characterization of HbMT2a, a metallothionein gene from Hevea brasiliensis Muell. Arg differently responds to abiotic stress and heavy metals.

作者信息

Li Yan, Chen Yue Yi, Yang Shu Guang, Tian Wei Min

机构信息

Rubber Research Institute, Chinese Academy of Tropical Agricultural Sciences, Danzhou, Hainan 571737, China.

Rubber Research Institute, Chinese Academy of Tropical Agricultural Sciences, Danzhou, Hainan 571737, China.

出版信息

Biochem Biophys Res Commun. 2015 May 22;461(1):95-101. doi: 10.1016/j.bbrc.2015.03.175. Epub 2015 Apr 6.

Abstract

Metallothioneins (MTs) are of low molecular mass, cysteine-rich proteins. They play an important role in the detoxification of heavy metals and homeostasis of intracellular metal ions, and protecting against intracellular oxidative damages. In this study a full-length cDNA of type 2 plant metallothioneins, HbMT2a, was isolated from 25 mM Polyethyleneglycol (PEG) stressed leaves of Hevea brasiliensis by RACE. The HbMT2a was 372bp in length and had a 237bp open reading frame (ORF) encoding for a protein of 78 amino acid residues with molecular mass of 7.772 kDa. The expression of HbMT2a in the detached leaves of rubber tree clone RY7-33-97 was up-regulated by Me-JA, ABA, PEG, H2O2, Cu(2+) and Zn(2+), but down-regulated by water. The role of HbMT2a protein in protecting against metal toxicity was demonstrated in vitro. PET-28a-HbMT2-beared Escherichia coli. Differential expression of HbMT2a upon treatment with 10 °C was observed in the detached leaves of rubber tree clone 93-114 which is cold-resistant and Reken501 which is cold-sensitive. The expression patterns of HbMT2a in the two rubber tree clones may be ascribed to a change in the level of endogenous H2O2.

摘要

金属硫蛋白(MTs)是低分子量、富含半胱氨酸的蛋白质。它们在重金属解毒、细胞内金属离子稳态以及抵御细胞内氧化损伤方面发挥着重要作用。在本研究中,通过RACE技术从25 mM聚乙二醇(PEG)胁迫的巴西橡胶树叶片中分离出2型植物金属硫蛋白的全长cDNA,即HbMT2a。HbMT2a长度为372bp,具有一个237bp的开放阅读框(ORF),编码一个由78个氨基酸残基组成、分子量为7.772 kDa的蛋白质。橡胶树无性系RY7 - 33 - 97离体叶片中HbMT2a的表达受茉莉酸甲酯(Me - JA)、脱落酸(ABA)、PEG、过氧化氢(H2O2)、铜离子(Cu(2+))上调,但受水下调。HbMT2a蛋白在体外具有抵御金属毒性的作用。携带PET - 28a - HbMT2的大肠杆菌。在耐寒的橡胶树无性系93 - 114和冷敏感的Reken501的离体叶片中,观察到10°C处理后HbMT2a的差异表达。两个橡胶树无性系中HbMT2a的表达模式可能归因于内源性H2O2水平的变化。

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