Lin M, Chang C J, Green N S
Department of Cell Biology, Albert Einstein College of Medicine, Bronx, NY 10461, USA.
Mutat Res. 1996 Apr 13;351(2):105-16. doi: 10.1016/0027-5107(95)00209-x.
Fluctuation analysis allows for the determination of mutation rates in cell cultures in vitro. As originally described by Luria and Delbruck and extended by Lea and Coulson and by Capizzi and Jameson, this analysis has been useful in estimating mutation rates in cultured cells where the frequency of mutational events is low. However. in cultures where high mutation rates and multiple independent mutation events occur, leading to the accumulation of many mutant cells, these standard methods may not apply. Here, we present a new method for the estimation of mutation rates based on the assumption that multiple events may contribute to the accumulation of mutant cells. We compared mutation rates determined by Lea and Coulson's and by Capizzi and Jameson's methods with those determined by our method using experimental and stimulated data from our studies of immunoglobulin gene mutation and isotype switching in B lymphocyte cultures. The three methods resulted in very different calculated rates when many mutants were present in the culture, such as when mutation rates were high, while only small differences in calculated rates were found when mutants were rare. Unlike previous fluctuation analysis calculations, our method is applicable for the estimation of both low and high rates.
波动分析可用于确定体外细胞培养中的突变率。正如最初由卢里亚和德尔布吕克所描述，并由利和库尔森以及卡皮齐和詹姆森扩展的那样，这种分析在估计突变事件频率较低的培养细胞中的突变率时很有用。然而，在高突变率和多个独立突变事件发生，导致许多突变细胞积累的培养物中，这些标准方法可能不适用。在此，我们基于多个事件可能导致突变细胞积累的假设，提出了一种估计突变率的新方法。我们将利和库尔森的方法以及卡皮齐和詹姆森的方法所确定的突变率与我们的方法所确定的突变率进行了比较，使用了我们对B淋巴细胞培养中的免疫球蛋白基因突变和同种型转换研究的实验数据和模拟数据。当培养物中存在许多突变体时，例如当突变率很高时，这三种方法得出的计算速率差异很大，而当突变体很少时，计算速率的差异很小。与以前的波动分析计算不同，我们的方法适用于估计低突变率和高突变率。
