Loveday K S, Latt S A
Nucleic Acids Res. 1978 Nov;5(11):4087-104. doi: 10.1093/nar/5.11.4087.
Chinese hamster ovary cells (CHO) grown for one cycle in bromodeoxyuridine (BrdU) contain a small amount (0.5%) of unusually dense double stranded DNA. This dense DNA has been previously interpreted as being bifilarly substituted with BrdU and hence evidence that sister chromatid exchange (SCE) formation proceeds via the Holliday model of recombination. However, the amount of this dense DNA is 100 times greater than that expected based on the SCE frequency in similarly cultured CHO cells, and it is not increased by treating the cells with mitomycin C. Moreover, contrary to expectations for bifilary substituted DNA, the amount of this dense DNA is not reduced by growing BrdU-labeled cells for a second cycle in TdR. Finally, DNA isolated from CHO cells contains a minor band (0.5%) with a density 0.025 gm/cc greater than that of the main band, whether or not BrdU has been incorporated. These results call into question the identification of this unusually dense DNA as bifilarly substituted and hence its previously postulated relationship to SCE formation.
在溴脱氧尿苷(BrdU)中培养一个周期的中国仓鼠卵巢细胞(CHO)含有少量(0.5%)异常致密的双链DNA。这种致密DNA先前被解释为被BrdU双股取代,因此证明姐妹染色单体交换(SCE)的形成是通过霍利迪重组模型进行的。然而,这种致密DNA的量比基于类似培养的CHO细胞中的SCE频率预期的量高100倍,并且用丝裂霉素C处理细胞不会增加其数量。此外,与对双股取代DNA的预期相反,通过在胸苷(TdR)中让BrdU标记的细胞进行第二个周期的培养,这种致密DNA的量不会减少。最后,无论是否掺入了BrdU,从CHO细胞中分离的DNA都含有一条次要条带(0.5%),其密度比主要条带的密度大0.025克/立方厘米。这些结果使人质疑将这种异常致密的DNA鉴定为双股取代,以及因此其先前假定的与SCE形成的关系。