Quantification of 8-OxodGuo Lesions in Double-Stranded DNA Using a Photoelectrochemical DNA Sensor
文献类型：期刊论文
| 作者 | Zhang, Bintian ; Guo, Liang-Hong ; Greenberg, Marc M. |
| 刊名 | ANALYTICAL CHEMISTRY
 |
| 出版日期 | 2012 |
| 卷号 | 84期号:14页码:6048－6053 |
| ISSN号 | 0003-2700 |
| 中文摘要 | Exposure of DNA to oxidative stress conditions results in the generation of 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodGuo). 8-OxodGuo is genotoxic if left unrepaired. We quantified 8-oxodGuo lesions in double-stranded DNA films by using a photoelectrochemical DNA sensor in conjunction with a specific covalent labeling method. A lesion-containing DNA film was assembled on a SnO2 nanoparticle modified indium tin oxide electrode through layer-by-layer electrostatic adsorption. The lesions were covalently labeled with a biotin conjugated spermine derivative, and ruthenium tris(bipyridine) labeled streptavidin was introduced as the signal reporter, molecule. Photocurrent increased with the number of lesions in the strand and decreased as the film was diluted with intact DNA. Quantification of 8-oxodGuo was achieved with an estimated detection limit of similar to 1 lesion in 650 bases or 1.6 fmol of 8-oxodGuo on the electrode. Incubation of the film with a DNA base excision repair enzyme, E. coli formamidopyrimidine DNA glycosylase (Fpg), resulted in complete loss of the signal, indicating efficient excision Of the isolated lesions in the nucleotide. Oxidatively generated DNA damage to a double stranded calf thymus DNA film by the Fenton reaction was then assessed. One 8-oxodGuo lesion in 520 bases was detected in DNA exposed to 50 mu M Fe2+/200 mu M H2O2. Treatment with Fpg reduced the photocurrent by 50%, indicating only partial excision of 8-oxodGuo. This suggests that tandem lesions, which are resistant to Fpg excision, are generated by the Fenton reaction. Unlike repair enzyme dependent methods, the sensor recognizes 8-oxodGuo in tandem lesions and can avoid underestimating DNA damage. |
| WOS记录号 | WOS:000306441200032 |
| 公开日期 | 2014-11-04 |
| 源URL | [http://ir.rcees.ac.cn/handle/311016/7743]  |
| 专题 | 生态环境研究中心_环境化学与生态毒理学国家重点实验室 |
| 推荐引用方式 GB/T 7714 | Zhang, Bintian,Guo, Liang-Hong,Greenberg, Marc M.. Quantification of 8-OxodGuo Lesions in Double-Stranded DNA Using a Photoelectrochemical DNA Sensor[J]. ANALYTICAL CHEMISTRY,2012,84(14):6048－6053. |
| APA | Zhang, Bintian,Guo, Liang-Hong,&Greenberg, Marc M..(2012).Quantification of 8-OxodGuo Lesions in Double-Stranded DNA Using a Photoelectrochemical DNA Sensor.ANALYTICAL CHEMISTRY,84(14),6048－6053. |
| MLA | Zhang, Bintian,et al."Quantification of 8-OxodGuo Lesions in Double-Stranded DNA Using a Photoelectrochemical DNA Sensor".ANALYTICAL CHEMISTRY 84.14(2012):6048－6053. |
入库方式： OAI收割
来源：生态环境研究中心
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