An Easy-To-Use Simulation Program Demonstrates Variations in Bacterial Cell Cycle Parameters Depending on Medium and Temperature 英文参考文献.docVIP

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An Easy-To-Use Simulation Program Demonstrates Variations in Bacterial Cell Cycle Parameters Depending on Medium and Temperature 英文参考文献.doc

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An Easy-To-Use Simulation Program Demonstrates Variations in Bacterial Cell Cycle Parameters Depending on Medium and Temperature 英文参考文献

AnEasy-To-UseSimulationProgramDemonstrates VariationsinBacterialCellCycleParametersDepending onMediumandTemperature CarolineStokke¤,IngvildFla?tten,KirstenSkarstad* DepartmentofCellBiology,InstituteforCancerResearch,TheNorwegianRadiumHospital,OsloUniversityHospital,Oslo,Norway Abstract ManystudiesareperformedonchromosomereplicationandsegregationinEscherichiacoliandotherbacteriacapableof 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. FlowcytometryisanimportanttoolforgenerationofexperimentalDNAdistributionsofcellpopulations.Here,a VisualBasicbasedsimulationprogramwaswrittenforthecomputationoftheoreticalDNAdistributionsfordifferentchoices ofcellcycleparameters(CandDphasedurations,doublingtimeetc).Thesecellcycleparameterscanbeiterateduntilthe 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 temperatureconditions,withfollowingmeasurementsbyflowcytometryandsimulationoftheDNAdistributions.Agood fitwasfoundforeachgrowthconditionbyuseofoursimulationprogram.Theresultingcellcycleparametersdisplayed clear inter-media differences in replication patterns, but indicated a high degree of temperature independence for each medium.Theexceptionwasthepoorestmedium(acetate),wherethecellsgrewwithoverlappingreplicationcyclesat42uC, butwithoutatthelowertemperatures. Wehavedevelopedaneasy-to-usetoolfordeterminationofbacteria’scellcycle parameters, and consequently the cells’ chromosome configurations. The procedure only requires DNA distribution measurementsbyflowcytometry.UseofthissimulationprogramforE.coliculturesshowsthatevencellsgrowingquite slowlycanhaveoverlappingreplicationcycles.Itisthereforealwaysimp