Synthesis and Characterization of DNA Fluorescent Probes Containing a Single Site-Specific Stereoisomer of anti-Benzo[a]pyrene Diol Epoxide-N-2-dG

CORC > 生态环境研究中心 > 中国科学院生态环境研究中心 > 环境化学与生态毒理学国家重点实验室

	Synthesis and Characterization of DNA Fluorescent Probes Containing a Single Site-Specific Stereoisomer of anti-Benzo[a]pyrene Diol Epoxide-N-2-dG
	Wang, Chao; Feng, Feng; Wang, Zhixin; Li, Tao; Le, X. Chris; Wang, Hailin
刊名	CHEMICAL RESEARCH IN TOXICOLOGY
	2009-04
卷号	22 期号:4 页码:676-682
中文摘要	We present a comprehensive study of synthesis and characterization of DNA probes containing covalently bound benzo[a]pyrene diol epoxide (BPDE) isomers at a defined site. Short oligonucleotides of 16mers containing a single trans-(+)- or trans-(-)-anti-BPDE-N-2-guanine adducts (BPDE-16mer) were first synthesized and then ligated with a fluorescently labeled single-stranded oligonucleotide. The ligation products (double-stranded or single-stranded 90mers) contained a single BPDE adduct of defined stereochemistry and a fluorescent label. The BPDE adduct could be recognized by a specific antibody, and the fluorescent label was useful for highly sensitive laser-induced fluorescence detection. The incorporation of single BPDE in the 16mers was validated by liquid chromatography, UV spectroscopy, and tandem mass spectrometry analysis. The stereochemistry of the single BPDE adducts in the 16mers was further identified by enzyme digestion-coupled stereoselective chromatography analysis. The ligation of BPDE-16mer with normal oligonucleotides for the synthesis of tetramethylrhodamine (TMR)-BPDE-90mers was evaluated. It was found that the modification of the 16mer by anti-BPDE could significantly reduce the ligation yield of ds90mer and lead to the formation of gapped DNA. The incorporation of a single anti-BPDE adduct in ligated ds90mers was confirmed using an antibody specific to the anti-BPDE-dG and affinity capillary electrophoresis. The detection limits of the TMR-BPDE-90mers by capillary electrophoresis coupled with laser-induced fluorescence are below 4 x 10(-19) mol.
WOS记录号	WOS:000265412300011
内容类型	期刊论文
源URL	[http://ir.rcees.ac.cn/handle/311016/21574]
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	Wang, Chao,Feng, Feng,Wang, Zhixin,et al. Synthesis and Characterization of DNA Fluorescent Probes Containing a Single Site-Specific Stereoisomer of anti-Benzo[a]pyrene Diol Epoxide-N-2-dG[J]. CHEMICAL RESEARCH IN TOXICOLOGY,2009,22(4):676-682.
APA	Wang, Chao,Feng, Feng,Wang, Zhixin,Li, Tao,Le, X. Chris,&Wang, Hailin.(2009).Synthesis and Characterization of DNA Fluorescent Probes Containing a Single Site-Specific Stereoisomer of anti-Benzo[a]pyrene Diol Epoxide-N-2-dG.CHEMICAL RESEARCH IN TOXICOLOGY,22(4),676-682.
MLA	Wang, Chao,et al."Synthesis and Characterization of DNA Fluorescent Probes Containing a Single Site-Specific Stereoisomer of anti-Benzo[a]pyrene Diol Epoxide-N-2-dG".CHEMICAL RESEARCH IN TOXICOLOGY 22.4(2009):676-682.