A novel method for site specific introduction of single model oxidative DNA lesions into oligodeoxyribonucleotides.
A novel method for site specific introduction of single model oxidative DNA lesions into oligodeoxyribonucleotides.
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DOI：
10.1093/nar/21.7.1563
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发表时间：
1993-04
影响因子：
14.9
通讯作者：
Z. Hatahet;A. Purmal;S. Wallace
中科院分区：
文献类型：
--
作者：
Z. Hatahet;A. Purmal;S. Wallace
小牛胸腺末端脱氧核苷酸转移酶用于将几种氧化碱基损伤产物掺入寡脱氧核苷酸的3'末端。在本报告所述的规定条件下，将二氢胸腺嘧啶、β-脲基异丁酸、胸腺嘧啶乙二醇、尿素、7-氢-8-氧代腺嘌呤、7-氢-8-氧代鸟嘌呤、5-羟基胞嘧啶和5-羟基尿嘧啶的单个残基掺入不同长度的寡脱氧核糖核苷酸中。该反应是有效的和成本有效的。反应产物的3'末端是噬菌体T4 DNA连接酶的合适底物，大大促进了含有独特的和位点特异性氧化DNA损伤的寡脱氧核苷酸的构建。
Calf thymus terminal deoxynucleotidyl transferase was used to incorporate several products of oxidative base damage onto the 3' end of oligodeoxyribonucleotides. Under the defined conditions described in this report, single residues of dihydrothymine, beta-ureidoisobutyric acid, thymine glycol, urea, 7-hydro-8-oxoadenine, 7-hydro-8-oxoguanine, 5-hydroxycytosine and 5-hydroxyuracil were incorporated into oligodeoxyribonucleotides of different lengths. The reaction is both efficient and cost effective. The 3' termini of the reaction products were suitable substrates for ligation by phage T4 DNA ligase, facilitating greatly the construction of oligodeoxyribonucleotides containing unique and site specific oxidative DNA lesions.