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Precise determinations of C and D periods by flow cytometry in Escherichia coli K-12 and B/r

¹ Section of Molecular Microbiology, BioCentrum – DTU, Technical University of Denmark – DTU, Building 301, DK-2800 Lyngby, Denmark
² Department of Microbiology, E. C. Slater Institute, BioCentrum Amsterdam, University of Amsterdam, 1018 WS Amsterdam, The Netherlands

Correspondence
Ole Michelsen
Ole.Michelsen{at}BioCentrum.dtu.dk

The C and D cell cycle periods of seven Escherichia coli K-12 strains and three E. coli B/r strains were determined by computer simulation of DNA histograms obtained by flow cytometry of batch cultures grown at several different generation times. To obtain longer generation times two of the K-12 strains were cultivated at several different dilution rates in glucose-limited chemostats. The replication period (C period) was found to be similar in K-12 and B/r strains grown at similar generation times. At generation times below 60 min the C period was constant; above 60 min it increased linearly with increasing generation time. The period from termination of replication to cell division (D period) was more variable. It was much shorter in B/r than in K-12 strains. Like the C period it was relatively constant at generation times below 60 min and it increased with increasing generation times at longer generation times. In glucose-limited chemostats good correlation was found between D periods and generation times, whereas batch cultures exhibited carbon-source-dependent variations. Chemostat cultures showed cell cycle variations very similar to those obtained in batch cultures. These flow cytometric determinations of cell cycle periods confirm earlier determinations of the C period and establish that the D period also varies with generation time in slowly growing cultures. In addition they extend the range of growth rates at which cell cycle periods have been determined in E. coli K-12.

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