An easy-to-use simulation program demonstrates variations in bacterial cell cycle parameters depending on medium and temperature.
An easy-to-use simulation program demonstrates variations in bacterial cell cycle parameters depending on medium and temperature.
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DOI：
10.1371/journal.pone.0030981
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Skarstad K
中科院分区：
文献类型：
--
作者：
Stokke C;Flåtten I;Skarstad K
在大肠杆菌和其他能够进行复杂复制的细菌中，人们对染色体复制和分离进行了许多研究，C期跨越几代。对于这类调查，了解复制模式，包括起始点和复制叉的拷贝数，对于正确解释结果至关重要。流式细胞术是产生实验细胞群体DNA分布的重要工具。在此，我们编写了一个基于VB的模拟程序，用于计算不同细胞周期参数(C、D期持续时间、倍增时间等)下的理论DNA分布。这些细胞周期参数可以迭代，直到获得实验和理论DNA直方图之间的最佳匹配。附件中包含仿真软件的Excel文件作为支持信息。在12种不同的培养基和温度条件下培养大肠杆菌，用流式细胞仪测量和模拟DNA分布。通过使用我们的模拟程序，发现每种生长条件都有很好的拟合。由此得到的细胞周期参数在复制模式上显示出明显的介质间差异，但表明每种介质的温度独立性很高。例外是最差的培养基(醋酸盐)，在42°C时，细胞以重叠的复制周期生长，但在较低的温度下没有。我们已经开发了一种易于使用的工具，用于确定细菌的细胞周期参数，从而确定细胞的染色体配置。该程序只需要通过流式细胞仪测量DNA分布。对大肠杆菌培养的模拟程序的使用表明，即使细胞生长得非常慢，也可能有重叠的复制周期。因此，不仅要假设细胞的复制模式，而且要在改变生长条件时实际确定细胞周期参数，这总是很重要的。
Many studies are performed on chromosome replication and segregation in Escherichia coli and other bacteria capable of complex replication with C phases spanning several generations. For such investigations an understanding of the replication patterns, including copy numbers of origins and replication forks, is crucial for correct interpretation of the results. Flow cytometry is an important tool for generation of experimental DNA distributions of cell populations. Here, a Visual Basic based simulation program was written for the computation of theoretical DNA distributions for different choices of cell cycle parameters (C and D phase durations, doubling time etc). These cell cycle parameters can be iterated until the best fit between the experimental and theoretical DNA histograms is obtained. The Excel file containing the simulation software is attached as supporting information. Cultures of Escherichia coli were grown at twelve different media and temperature conditions, with following measurements by flow cytometry and simulation of the DNA distributions. A good fit was found for each growth condition by use of our simulation program. The resulting cell cycle parameters displayed clear inter-media differences in replication patterns, but indicated a high degree of temperature independence for each medium. The exception was the poorest medium (acetate), where the cells grew with overlapping replication cycles at 42°C, but without at the lower temperatures. We have developed an easy-to-use tool for determination of bacteria's cell cycle parameters, and consequently the cells' chromosome configurations. The procedure only requires DNA distribution measurements by flow cytometry. Use of this simulation program for E. coli cultures shows that even cells growing quite slowly can have overlapping replication cycles. It is therefore always important not only to assume cells' replication patterns, but to actually determine the cell cycle parameters when changing growth conditions.
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影响因子：
3.2
作者：
Bipatnath, M;Dennis, PP;Bremer, H
通讯作者：
Bremer, H
影响因子：
5.6
作者：
HELMSTETTER, CE;PIERUCCI, O
通讯作者：
PIERUCCI, O
影响因子：
2.6
作者：
BOYE, E;LOBNEROLESEN, A
通讯作者：
LOBNEROLESEN, A
影响因子：
3.7
作者：
Jannière L;Canceill D;Suski C;Kanga S;Dalmais B;Lestini R;Monnier AF;Chapuis J;Bolotin A;Titok M;Le Chatelier E;Ehrlich SD
通讯作者：
Ehrlich SD
影响因子：
2.1
作者：
Adachi, Shun;Fukushima, Toru;Hiraga, Sota
通讯作者：
Hiraga, Sota