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2007•Journal of Nuclear Agricultural SciencesRequires access

DISRUPTION AND CHARACTERIZATION OF THE EXCISION REPAIR PATHWAY IN THE EXTREMELY RADIORESISTANT BACTERIUM Deinococcus SP.BR501

Liu Xiu-min, Jing Wu, Ming Chen

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Abstract

Deinococcus sp.BR501,an extremely radioresistant bacterium may contain two nucleotide excision repair pathways: the UV damage endonuclease β(UvsE)-dependent excision repair pathway and the UvrABC-dependent pathway.And the UvsE(coded by dr1819) and UvrABC(Unit A coded by dr1771) are their key enzymes respectively.PCR primers were designed and homologous genes were cloned and disrupted in vitro according to the completely nucleotide sequence of Deinococcus radiodurans R1 genome.Then PCR production was transformed to BR501,and the disrupted mutants(△dr1771,△dr1819 and △dr1771dr1819) were checked and confirmed by homologous recombination.These mutants and the wild type were irradiated by UV light and exposed to the DNA-damaging agents MMC and H_2O_2.The results showed that these pathways were existed in BR501 and only the two pathway losses could result in increased sensitivity to UV and MMC.

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What this paper is about

Deinococcus sp.BR501,an extremely radioresistant bacterium may contain two nucleotide excision repair pathways: the UV damage endonuclease β(UvsE)-dependent excision repair pathway and the UvrABC-dependent pathway.And the UvsE(coded by dr1819) and UvrABC(Unit A coded by dr1771) are their key enzymes respectively.PCR primers were designed and homologous genes were cloned and disrupted in vitro according to the completely nucleotide sequence of Deinococcus radiodurans R1 genome.Then PCR production was transformed to BR501,and the disrupted mutants(△dr1771,△dr1819 and △dr1771dr1819) were checked and confirmed by homologous recombination.These mutants and the wild type were irradiated by UV light and exposed to the DNA-damaging agents MMC and H_2O_2.The results showed that these pathways were existed in BR501 and only the two pathway losses could result in increased sensitivity to UV and MMC.

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Available abstract

Deinococcus sp.BR501,an extremely radioresistant bacterium may contain two nucleotide excision repair pathways: the UV damage endonuclease β(UvsE)-dependent excision repair pathway and the UvrABC-dependent pathway.And the UvsE(coded by dr1819) and UvrABC(Unit A coded by dr1771) are their key enzymes respectively.PCR primers were designed and homologous genes were cloned and disrupted in vitro according to the completely nucleotide sequence of Deinococcus radiodurans R1 genome.Then PCR production was transformed to BR501,and the disrupted mutants(△dr1771,△dr1819 and △dr1771dr1819) were checked and confirmed by homologous recombination.These mutants and the wild type were irradiated by UV light and exposed to the DNA-damaging agents MMC and H_2O_2.The results showed that these pathways were existed in BR501 and only the two pathway losses could result in increased sensitivity to UV and MMC.

Key concepts: Deinococcus radiodurans, DNA repair, Nucleotide excision repair, Deinococcus, Homologous recombination, Biology, Mutant, DNA damage

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DISRUPTION AND CHARACTERIZATION OF THE EXCISION REPAIR PATHWAY IN THE EXTREMELY RADIORESISTANT BACTERIUM Deinococcus SP.BR501 — Research Paper | ScholarLens