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电离辐射引起的DNA双链断裂是细胞最重要的生物损伤之一。但是中性蔗糖沉降法及最近兴起的中性滤过洗脱法一般不适于哺乳动物双链断裂及细胞内修复的研究。作者发明了一种以病毒作为细胞内探针的方法来研究单、双链断裂。此类病毒是一种亲核的小型环状DNA病毒,自身缺乏DNA修复能力(papovaviruses SV40 and BK)。将受照病毒感染未受照细胞后分别进行不同时间的培养。分离核,提取病毒DNA并测定其剩余的损伤。此法用琼脂糖凝胶电泳分离病毒DNA,可见三条带,分别含双链断裂,单链断裂及无断裂的DNA分子。Southern blot法
Ionizing radiation-induced DNA double-strand breaks are one of the cell’s most important biological lesions. However, neutral sucrose sedimentation and the recent rise of the neutral filtration elution method are generally not suitable for mammalian double-strand breaks and intracellular repair studies. The authors invented a virus as an intracellular probe to study single and double strand breaks. The virus is a nucleophilic, small circular DNA virus that itself lacks the DNA repair capacity (papovaviruses SV40 and BK). Will be infected with virus-infected cells were cultured at different times. Nuclei were isolated, viral DNA was extracted and the remaining damage was measured. This method agarose gel electrophoresis separation of viral DNA, showing three bands, containing double-strand breaks, single-strand breaks and non-breaking DNA molecules. Southern blot method