Research project P 14591 Analysis of cellular Physiology of recombinant E. coli cells Nicole BORTH 26.6.2000
Strong overexpression of recombinant proteins in bacteria exert a metabolic burden on the host cell. As cell
populations usually do not react homogeneously to such stress situations, a clearer picture of the physiological
consequences can be obtained with flow cytometry, where several parameters can be measured simultaneously in
single cells and the distribution of these parameters, and their combinations, can be visualized. During the project P
1227 1 -MOB, methods have been developed for the simultaneous measurement of total DNA, total protein and
specific recombinant protein. These methods require that the cells are fixed prior to staining, so that no data on cell
vitality can be obtained.
The aim of this project extension is now to analyze the development of cell vitality as a consequence of metabolic
burden due to recombinant protein overproduction. The major answers that are expected are how vitality changes in
time and distribution within the culture, and how vitality is related to other parameters like cell size and DNA
content.