Macleod H, Stadler D
Mol Gen Genet. 1986 Feb;202(2):321-6. doi: 10.1007/BF00331658.
Germinated conidia of Neurospora have been monitored for their ability to excise pyrimidine dimers. Dimer concentration was measured in DNA extracted immediately after UV treatment, and it was compared to that of DNA from cells which had a post-UV incubation before extraction. Two methods were used to assay dimer level in DNA: measurement of the number of single-strand breaks (as revealed in alkaline sucrose gradients) produced by a dimer-specific endonuclease; monitoring the ability to compete for binding to dimer-specific antibodies in a radioimmunoassay. Both methods showed efficient excision of dimers by wild-type and by uvs-2, even though an earlier study had reported that uvs-2 was unable to excise dimers. UV-induced mutation shows a dose-rate effect: acute UV yields several times as many mutations as does the same dose of chronic UV. There is a parallel effect on dimer accumulation. The concentration of dimers at the conclusion of the UV treatment shows a strong correlation with the resultant mutation frequency.
人们监测了粗糙脉孢菌发芽的分生孢子切除嘧啶二聚体的能力。在紫外线处理后立即提取的DNA中测量二聚体浓度,并将其与在提取前经过紫外线后培养的细胞的DNA中的二聚体浓度进行比较。使用两种方法测定DNA中的二聚体水平:测量由二聚体特异性内切核酸酶产生的单链断裂数(如在碱性蔗糖梯度中所示);在放射免疫测定中监测与二聚体特异性抗体结合的竞争能力。两种方法均显示野生型和uvs-2能有效切除二聚体,尽管早期研究报告称uvs-2无法切除二聚体。紫外线诱导的突变表现出剂量率效应:急性紫外线产生的突变数量是相同剂量慢性紫外线产生的突变数量的几倍。对二聚体积累也有类似的影响。紫外线处理结束时二聚体的浓度与产生的突变频率密切相关。