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来自绿藻门(绿藻纲,共球藻目)的铁超氧化物歧化酶基因的分子特征及表达分析

Molecular characterization and expression analysis of iron superoxide dismutase gene from (Trebouxiophyceae, Chlorophyta).

作者信息

Ismaiel Mostafa M S, Piercey-Normore Michele D

机构信息

1Department of Biological Sciences, Faculty of Science, University of Manitoba, Winnipeg, MB R3T 2N2 Canada.

2Botany Department, Faculty of Science, Zagazig University, Zagazig, 44519 Egypt.

出版信息

Physiol Mol Biol Plants. 2019 Jan;25(1):221-228. doi: 10.1007/s12298-018-0569-5. Epub 2018 Jun 18.

Abstract

The FeSOD isoforms of were identified, a preliminary characterization of the enzyme was conducted, and the relationship among the FeSOD gene from and that of other organisms was examined. The FeSOD has an open reading frame of 612 bp that encodes 203 deduced amino acids with a molecular mass of 23 kDa. Expression of the recombinant FeSOD gene was done successfully in . The purified FeSOD has a specific enzyme activity that reached 688 U mg protein (in vitro assay). Alkaline conditions showed the highest activity for the recombinant FeSOD. Moreover, it showed a relative thermostability up to 50 °C, while at 50 and 70 °C, the activity was reduced by 32 and 68%, respectively, after 1 h as compared to the maximum. Phylogenetic analysis revealed three main clusters i.e., the prokaryotic Cyanophyta, bacteria, and the eukaryotic Chlorophyta intermingled with plant species and a dinoflagellate. was closely grouped with , however, other species showed a relative disparity. Alignment of FeSOD gene sequences of the different species showed many conserved regions which could be used for FeSOD sequences among unexplored species and may be useful for the taxonomy of the revised coccoid species.

摘要

鉴定了[具体物种]的铁超氧化物歧化酶(FeSOD)同工型,对该酶进行了初步表征,并研究了[具体物种]的FeSOD基因与其他生物的FeSOD基因之间的关系。FeSOD具有一个612 bp的开放阅读框,编码203个推导氨基酸,分子量为23 kDa。重组FeSOD基因在[具体宿主]中成功表达。纯化后的FeSOD的比酶活性在体外测定中达到688 U/mg蛋白质。碱性条件下重组FeSOD的活性最高。此外,它在高达50°C时表现出相对热稳定性,而在50°C和70°C时,与最高活性相比,1小时后活性分别降低了32%和68%。系统发育分析揭示了三个主要类群,即原核蓝藻、细菌以及与植物物种和一种甲藻混合的真核绿藻。[具体物种]与[另一具体物种]紧密聚类,然而,其他物种表现出相对差异。不同物种的FeSOD基因序列比对显示出许多保守区域,这些区域可用于未探索物种的FeSOD序列分析,可能对修订后的球状体[具体物种]的分类学有用。

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