Analysis of cellular Physiology of recombinant E coli cells
Abstract
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 P12271-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.
Schlagworte Durchflusszytometrie rekombinante Bakterien Hochproduzenten Stress
Publikationen
Mitarbeiter*innen
Nicole Borth
Ao.Univ.Prof. Dipl.-Ing. Dr.nat.techn. Nicole Borth
nicole.borth@boku.ac.at
Tel: +43 1 47654-79064, 79065
Projektleiter*in
01.09.2000 - 01.09.2001