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水稻金属硫蛋白异构体(OsMTI-1b)在酿酒酵母中的异源表达增强了对镉、过氧化氢和乙醇的耐受性。

Heterologous expression of a rice metallothionein isoform (OsMTI-1b) in Saccharomyces cerevisiae enhances cadmium, hydrogen peroxide and ethanol tolerance.

作者信息

Ansarypour Zahra, Shahpiri Azar

机构信息

Isfahan University of Technology, College of Agriculture, Department of Biotechnology, Isfahan, Iran.

Isfahan University of Technology, College of Agriculture, Department of Biotechnology, Isfahan, Iran.

出版信息

Braz J Microbiol. 2017 Jul-Sep;48(3):537-543. doi: 10.1016/j.bjm.2016.10.024. Epub 2017 Feb 4.

Abstract

Metallothioneins are a superfamily of low-molecular-weight, cysteine (Cys)-rich proteins that are believed to play important roles in protection against metal toxicity and oxidative stress. The main purpose of this study was to investigate the effect of heterologous expression of a rice metallothionein isoform (OsMTI-1b) on the tolerance of Saccharomyces cerevisiae to Cd, HO and ethanol stress. The gene encoding OsMTI-1b was cloned into p426GPD as a yeast expression vector. The new construct was transformed to competent cells of S. cerevisiae. After verification of heterologous expression of OsMTI-1b, the new strain and control were grown under stress conditions. In comparison to control strain, the transformed S. cerevisiae cells expressing OsMTI-1b showed more tolerance to Cd and accumulated more Cd ions when they were grown in the medium containing CdCl. In addition, the heterologous expression of GST-OsMTI-1b conferred HO and ethanol tolerance to S. cerevisiae cells. The results indicate that heterologous expression of plant MT isoforms can enhance the tolerance of S. cerevisiae to multiple stresses.

摘要

金属硫蛋白是一类低分子量、富含半胱氨酸(Cys)的蛋白质超家族,被认为在抵御金属毒性和氧化应激方面发挥着重要作用。本研究的主要目的是调查水稻金属硫蛋白异构体(OsMTI-1b)的异源表达对酿酒酵母耐受镉、过氧化氢和乙醇胁迫能力的影响。将编码OsMTI-1b的基因克隆到作为酵母表达载体的p426GPD中。将新构建体转化到酿酒酵母感受态细胞中。在验证OsMTI-1b的异源表达后,新菌株和对照在胁迫条件下生长。与对照菌株相比,表达OsMTI-1b的转化酿酒酵母细胞在含有CdCl的培养基中生长时,对镉表现出更高的耐受性并积累了更多的镉离子。此外,GST-OsMTI-1b的异源表达赋予酿酒酵母细胞对过氧化氢和乙醇的耐受性。结果表明,植物MT异构体的异源表达可以增强酿酒酵母对多种胁迫的耐受性。

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