Weinfeld M, Buchko G W
Radiobiology Program, Cross Cancer Institute, Edmonton, Alberta, Canada.
IARC Sci Publ. 1993(124):95-103.
This paper reviews the 32P-postlabelling techniques that have been employed in the detection and quantitation of apurinic sites and DNA damage induced by UV and ionizing radiation. The two major approaches utilize different enzymes for DNA digestion--in one case, micrococcal nuclease and calf spleen phosphodiesterase, and in the other DNase I, snake venom phosphodiesterase and calf intestinal phosphatase. As a result, each technique detects different classes of lesions with limited overlap and can, therefore, be considered complementary. The enzyme dependence and other technical aspects, in particular problems of background resulting from undamaged DNA, and some of the applications with cellular DNA and naked DNA, are discussed.
本文综述了用于检测和定量脱嘌呤位点以及紫外线和电离辐射诱导的DNA损伤的32P后标记技术。两种主要方法利用不同的酶进行DNA消化——一种情况下,使用微球菌核酸酶和小牛脾磷酸二酯酶,另一种情况下,使用DNase I、蛇毒磷酸二酯酶和小牛肠磷酸酶。因此,每种技术检测不同类别的损伤,重叠有限,因此可以认为是互补的。本文讨论了酶依赖性和其他技术方面,特别是未受损DNA产生的背景问题,以及一些在细胞DNA和裸露DNA方面的应用。