Talpaert-Borlé M, Liuzzi M
Biochim Biophys Acta. 1983 Sep 9;740(4):410-6. doi: 10.1016/0167-4781(83)90089-1.
A simple and rapid method is described for the determination of AP (apurinic/apyrimidinic) sites in DNA. The method involves the reaction of [14C]methoxyamine with the aldehyde group present in the deoxyribose moiety after a base loss. Studies with alkylated-depurinated DNA and with uracil-containing polydeoxyribonucleotides depyrimidinated by uracil-DNA glycosylase show that methoxyamine reacts with both apurinic and apyrimidinic sites in a rapid and exhaustive way. Under standard conditions (30-min incubation with 5 mM methoxyamine at 37 degrees C, pH 7.2) untreated DNA is almost unreactive and the [14C]methoxyamine incorporation in DNA is proportional to the number of AP sites. Since the methoxyamine reaction is free from any degradative effect on DNA, AP sites may be estimated from a simple determination of the acid-insoluble radioactivity.
本文描述了一种简单快速的测定DNA中脱嘌呤/脱嘧啶（AP）位点的方法。该方法包括在碱基丢失后，使[¹⁴C]甲氧基胺与脱氧核糖部分存在的醛基发生反应。对烷基化脱嘌呤DNA以及经尿嘧啶-DNA糖基化酶脱嘧啶的含尿嘧啶多脱氧核糖核苷酸的研究表明，甲氧基胺能快速且彻底地与脱嘌呤和脱嘧啶位点发生反应。在标准条件下（37℃、pH 7.2时与5 mM甲氧基胺孵育30分钟），未处理的DNA几乎不发生反应，且DNA中[¹⁴C]甲氧基胺的掺入量与AP位点的数量成正比。由于甲氧基胺反应对DNA没有任何降解作用，因此可通过简单测定酸不溶性放射性来估算AP位点。
