QSB: Variability in Cell Populations

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The overall objective of this project is to determine the relationship between phenotypic variations and the associated whole genome transcriptional pattern in populations of E.coli and S. cerevisiae cells cultivated under well defined, reproducible conditions. This will be done in three types of experiments distinguished according to the cultivation mode. The first type of experiments will involve simple batch cultivation of cells. Samples will be taken from different growth stages of the culture. The second type of experiments will involve continuous cultivation of cells in a chemostat operated at different steady states. The third type of experiments will involve exposure of the cells to an environmental challenge. Such challenges will include the shifting of a steady state chemostat culture to a new dilution rate. The response to such a nutritional upshift or downshift will be analyzed. In all types of experiments the phenotypic variability of the culture will be determined by flow cytometry and subpopulations of cells will be isolated via cell sorting. Sorted cells will be subjected to protocols for microarray analysis to determine the whole genome transcriptional profile. To make this possible an amplification protocol will be developed to reproducibly generate a sufficient amount of RNA from only a few cells to permit quantitative microarray analysis.