Hatahet Z, Purmal A A, Wallace S S
Department of Microbiology and Molecular Genetics, Markey Center for Molecular Genetics, University of Vermont, Burlington 05405.
Nucleic Acids Res. 1993 Apr 11;21(7):1563-8. doi: 10.1093/nar/21.7.1563.
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.
小牛胸腺末端脱氧核苷酸转移酶被用于将几种氧化碱基损伤产物掺入到寡脱氧核糖核苷酸的3'末端。在本报告所述的特定条件下，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被掺入到不同长度的寡脱氧核糖核苷酸中。该反应既高效又经济。反应产物的3'末端是噬菌体T4 DNA连接酶连接的合适底物，极大地促进了含有独特且位点特异性氧化DNA损伤的寡脱氧核糖核苷酸的构建。
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