The variability in protein number was studied. The reporter protein Enhanced Green Fluorescent Protein (EGFP) in various strains of Escherichia coli with regulated and unregulated plasmid copy number systems were investigated. The relaxation of perturbed population was studied in two different systems and cell division was observed to drive the relaxation of perturbed populations. It was found that an unregulated system has more inclusion bodies in comparison to regulated one. This accounted for the difference in correlation between cell size and protein number in two different systems. Flow cytometry was cross-validated with fluorescence microscopy and it was concluded that the Forward Scatter signal in a flow cytometer is representative of size in terms of generating size distributions. A new method for determining cell viability was devised, reducing the analysis time to less than five minutes. The unregulated system in comparison to the regulated one tends to have higher percentage of dead cells.
| Identifer | oai:union.ndltd.org:TORONTO/oai:tspace.library.utoronto.ca:1807/11162 |
| Date | 30 July 2008 |
| Creators | Sardana, Vandit |
| Contributors | McMillen, David |
| Source Sets | University of Toronto |
| Language | en_ca |
| Detected Language | English |
| Type | Thesis |
| Format | 12533303 bytes, application/pdf |
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