Krasin F, Hutchinson F
Biophys J. 1978 Dec;24(3):657-64. doi: 10.1016/S0006-3495(78)85411-3.
Supercircular gamma phage DNA with 10 bromouracils/100 thymine bases, irradiated with 313 nm light in Tris buffer and sedimented on alkaline and neutral gradients, showed 4.6 alkali-labile bonds per true single-strand break, in agreement with Hewitt and Marburger (1975 Photochem. Photobiol. 21:413). The same DNA irradiated in Escherichia coli host cells showed about the same number of breaks in alkaline gradients for equal fluence, but only 0.5 alkali-labile bond per true break. Similarly, E. coli DNA with bromouracil irradiated in the cells showed only 10--20% more breaks when denatured with 0.1 M NaOH than under neutral conditions with 9 M sodium perchlorate at 50 degrees C. These results show that true single-strand breaks occur more frequently than alkali-labile bonds after ultraviolet irradiation of DNA containing bromouracil in cells.
每100个胸腺嘧啶碱基含有10个溴尿嘧啶的超环状γ噬菌体DNA,在Tris缓冲液中用313nm光照射,然后在碱性和中性梯度中沉降,结果显示每一个真正的单链断裂有4.6个碱不稳定键,这与休伊特和马伯格(1975年,《光化学与光生物学》21:413)的研究结果一致。在大肠杆菌宿主细胞中照射的相同DNA,在相同通量下,碱性梯度中的断裂数大致相同,但每个真正的断裂只有0.5个碱不稳定键。同样,细胞中含有溴尿嘧啶的大肠杆菌DNA在用0.1M NaOH变性时,与在50℃下用9M高氯酸钠在中性条件下相比,断裂数仅多10%-20%。这些结果表明,在细胞中紫外线照射含溴尿嘧啶的DNA后,真正的单链断裂比碱不稳定键更频繁地发生。