Host cell reactivation of ozone-treated T3 bacteriophage by different strains ofEscherichia coli

L'Hérault, Pierre; Chung, Young Sup

doi:10.1007/bf01955788

Cited by 5 publications

(2 citation statements)

References 24 publications

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“…Base excision repair (BER) has also been analyzed by HCR, using either nonspecific methylating agents such as N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) (23) or very specific methylating agents known to produce substrates for specific BER glycosylases, such as 5-(3-methyl-1-triazeno)imidazole-4-carboxamide (MTIC) (24). BER is also known to remediate some of the damage caused by exposure of DNA to oxidative agents, such as ozone and sources of singlet oxygen (25,26). The steps in BER beyond the initial glycosylation can be analyzed by acid/heat treatment of the plasmid to produce apurinic sites (27), or apurinic sites can be specifically introduced (28).…”

Section: Notesmentioning

confidence: 99%

Analysis of DNA Repair Using Transfection-Based Host Cell Reactivation

Johnson

Latimer

Molecular Toxicology Protocols

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SummaryHost cell reactivation (HCR) is a transfection-based assay in which intact cells repair damage localized to exogenous DNA. This chapter provides instructions for the application of this technique using UV irradiation as a source of damage to a luciferase reporter plasmid. Through measurement of the activity of a reporter enzyme, the amount of damaged plasmid that a cell can "reactivate" or repair and express can be quantitated. Different DNA repair pathways can be analyzed by this technique by damaging the reporter plasmid in different ways. Because it involves repair of a transcriptionally active gene, when applied to UV damage the HCR assay measures the capacity of the host cells to perform transcription-coupled repair (TCR), a subset of the overall nucleotide excision repair pathway that specifically targets transcribed gene sequences.

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Section: Notesmentioning

confidence: 99%

Analysis of DNA Repair Using Transfection-Based Host Cell Reactivation

Johnson

Latimer

Molecular Toxicology Protocols

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“…BER is also known to remediate some of the damage caused by exposure of DNA to oxidative agents, such as ozone and sources of singlet oxygen [24, 25]. The steps in BER beyond the initial glycosylation can be analyzed by acid/heat treatment of the plasmid to produce apurinic sites [26] or apurinic sites can be specifically introduced [27].…”

mentioning

confidence: 99%

Analysis of Actively Transcribed DNA Repair Using a Transfection-Based System

Latimer

2014

Molecular Toxicology Protocols

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Host cell reactivation (HCR) is a transfection-based assay in which intact cells repair damage localized to exogenous DNA. This chapter provides instructions for the application of this technique, using as an exemplar UV irradiation as a source of damage to a luciferase reporter plasmid. Through measurement of the activity of a successfully transcribed and translated reporter enzyme, the amount of damaged plasmid that a cell can “reactivate” or repair and express can be quantitated. Different DNA repair pathways can be analyzed by this technique by damaging the reporter plasmid in different ways. Since it involves repair of a transcriptionally active gene, when applied to UV damage the HCR assay measures the capacity of the host cells to perform transcription-coupled repair, a subset of the overall nucleotide excision repair pathway that specifically targets transcribed gene sequences.

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