Isolation and characterization of oligodeoxynucleotides containing dG-N2-AAF and oxidation products of dG-C8-AF.
Isolation and characterization of oligodeoxynucleotides containing dG-N2-AAF and oxidation products of dG-C8-AF.
复制标题
含有 dG-N2-AAF 和 dG-C8-AF 氧化产物的寡脱氧核苷酸的分离和表征。
DOI：
10.1093/carcin/12.5.813
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发表时间：
1991
期刊：
影响因子：
4.7
通讯作者：
Grollman,AP
中科院分区：
文献类型：
--
作者：
Shibutani,S;Gentles,R;Johnson,F;Grollman,AP
分离和鉴定了十脱氧核苷酸containingN-(deoxyguanosine-N2-yl)-2-acetylaminofluorene(DG-N_2-AAF)和三个最近报道的n-脱氧鸟苷-8-基-2-氨基荧烯(DG-C_8-AF)的氧化产物。合成了DG-N_2-AAF，其结构经质谱学和核磁共振氢谱确证。通过允许d(CACTAGTCAC)与N-乙酰氧基-AAF反应并用高效液相色谱分离产物，制备了含有DG-N_2-AAF和DG-C_8-AAF的十脱氧核苷酸。将含有DG-C8-AAF的十聚体在好氧碱性条件下培养。在2-巯基乙醇存在下，加合物被脱乙酰化；在没有抗氧化剂的情况下，形成带有氧化产物的十聚体。用聚丙烯酰胺凝胶电泳法测定了修饰低聚物的均一性。这些修饰的寡脱氧核苷酸将被用于将DG-N-氨基荧加合物和定点氧化损伤引入DNA，从而将这些加合物与其突变特性相关联。
Decadeoxynudeotides containingN-(deoxyguanosine-N2-yl)-2-acetylaminofluorene (dG-N2-AAF) and three recently described products of oxidation ofN-{deoxyguanosine-8-yl)-2-aminofluorene (dG-C8-AF) were isolated and characterized. dG-N2-AAF was synthesized; its structure was established by mass spectroscopic and1H-NMR analysis. Decadeoxynudeotides containing dG-N2-AAF and dG-C8-AAF were prepared by permitting d(CACTAGTCAC) to react withN-acetoxy-AAF and separating the products by HPLC. The decamer containing dG-C8-AAF was incubated under aerobic alkaline conditions. In the presence of 2-mercaptoethanol, the adduct is deacetylated; in the absence of antioxidant, decamers bearing oxidation products are formed. Homogeneity of the modified oligomers was established by polyacrylamide gel electrophoresis. The modified oligodeoxy- nucleotides will be used to introduce dG-N2-aminofluorene adducts and oxidative lesions, site-specifically, into DNA, thereby to correlate these adducts with their mutagenic properties.