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嗜盐古菌盐沼盐杆菌的rad2基因在紫外线诱导的DNA光产物修复中发挥作用。

The rad2 gene of haloarchaeum Halobacterium salinarum is functional in the repair of ultraviolet light induced DNA photoproducts.

作者信息

Vafadarnejad Ehsan, Amoozgar Mohammad Ali, Khansha Javad, Fallahzade Ramin

机构信息

Extremophiles Laboratory, Department of Microbiology, Faculty of Biology, College of Science, University of Tehran, Tehran, Iran.

Extremophiles Laboratory, Department of Microbiology, Faculty of Biology, College of Science, University of Tehran, Tehran, Iran.

出版信息

Microbiol Res. 2015 Apr;173:44-9. doi: 10.1016/j.micres.2015.01.012. Epub 2015 Feb 7.

Abstract

There are a lot of bacterial and eukaryotic DNA repair gene homologs among sequenced archaeal genomes but there is little information about DNA repair mechanisms and the interaction of involved repair proteins. In order to study DNA repair mechanisms in the third domain of life, we studied these processes in the model archaeon, Halobacterium salinarum. H. salinarum has homologs of eukaryotic nucleotide excision repair genes such as rad2 gene. A functional analysis of rad2 was performed by knocking down of this gene. We introduced an antisense RNA expression vector into the cells and the sensitivity of transformants against ultraviolet light exposure was measured to determine whether rad2 gene performs any role in the repair of the DNA lesions induced by UV light or not. Our data suggests that rad2 is functional in this pathway and knocked down strains were unable to completely repair the UV induced DNA damages. In this study, for the first time antisense RNA is used for functional analysis of a gene in H. salinarum and it is shown that antisense RNA could be used as a reliable genetic tool for understanding of the archaeal genetics.

摘要

在已测序的古菌基因组中有许多细菌和真核生物的DNA修复基因同源物,但关于DNA修复机制以及相关修复蛋白的相互作用的信息却很少。为了研究生命第三域中的DNA修复机制,我们在模式古菌盐沼盐杆菌中研究了这些过程。盐沼盐杆菌具有真核生物核苷酸切除修复基因的同源物,如rad2基因。通过敲低该基因对rad2进行了功能分析。我们将反义RNA表达载体导入细胞,并测量转化体对紫外线照射的敏感性,以确定rad2基因在修复紫外线诱导的DNA损伤中是否发挥任何作用。我们的数据表明rad2在该途径中具有功能,敲低菌株无法完全修复紫外线诱导的DNA损伤。在本研究中,首次将反义RNA用于盐沼盐杆菌中基因的功能分析,结果表明反义RNA可作为理解古菌遗传学的可靠遗传工具。

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