Yeisley W G, Pollard E C
Biophys J. 1969 Jul;9(7):925-49. doi: 10.1016/S0006-3495(69)86427-1.
A method has been worked out for studying the division of single bacterial cells (Escherichia coli B/r) in a uniform environment. Under optimal conditions the daughters of one single cell are found to divide at different times, a fact which indicates that they are not identical. The spread in generation times can be estimated quantitatively. When cells are irradiated with gamma rays in nutrient broth there is an increase in the spread in generation times, and the number of three-cell progeny (which require considerable difference between the daughters) rises. The results are consistent with the idea that damage to a segment of DNA has taken place and that there are three growing points on the DNA at any one time. In nutrient broth there is some evidence for repair of damage. For cells irradiated in minimal medium the pattern is different. The increase in generation times is not so marked, and only slight increase in three-cell progeny is seen. The results suggest that there is the same class of damage to the DNA, with only one growing point present and less repair. Using the criterion that no increase in generation time at all is permitted, we can estimate the total escape probability after radiation. The resulting calculation of critical target size is much closer to the whole bacterial chromosome than is found from normal plating experiments. The behavior of cells that have been exposed to irradiated medium is quite different, involving very long lags and cell death.
已制定出一种方法来研究单个细菌细胞(大肠杆菌B/r)在均匀环境中的分裂情况。在最佳条件下,发现单个细胞的子代在不同时间进行分裂,这一事实表明它们并不相同。世代时间的差异可以进行定量估计。当细胞在营养肉汤中受到伽马射线照射时,世代时间的差异会增加,并且三细胞后代(这需要子代之间有相当大的差异)的数量会增加。这些结果与这样一种观点一致,即DNA的某一片段发生了损伤,并且在任何时候DNA上都有三个生长点。在营养肉汤中,有一些证据表明损伤得到了修复。对于在基本培养基中照射的细胞,情况则不同。世代时间的增加不那么明显,并且只观察到三细胞后代略有增加。结果表明,对DNA的损伤属于同一类型,只有一个生长点,且修复较少。使用完全不允许世代时间增加的标准,我们可以估计辐射后的总逃逸概率。由此计算出的关键靶标大小比通过正常平板实验得出的结果更接近整个细菌染色体。暴露于受照射培养基中的细胞行为则大不相同,会出现很长的延迟期和细胞死亡。