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dc.contributor.advisor Hellinga, Homme W en_US
dc.contributor.advisor You, Lingchong en_US
dc.contributor.author Marguet, Philippe Robert en_US
dc.date.accessioned 2011-01-06T16:04:00Z
dc.date.available 2011-01-06T16:04:00Z
dc.date.issued 2010 en_US
dc.identifier.uri http://hdl.handle.net/10161/3143
dc.description Dissertation en_US
dc.description.abstract <p>Driven by the development of new technologies and an ever expanding knowledge base of molecular and cellular function, Biology is rapidly gaining the potential to develop into a veritable engineering discipline - the so-called `era of synthetic biology' is upon us. Designing biological systems is advantageous because the engineer can leverage existing capacity for self-replication, elaborate chemistry, and dynamic information processing. On the other hand these functions are complex, highly intertwined, and in most cases, remain incompletely understood. Brazenly designing within these systems, despite large gaps in understanding, engenders understanding because the design process itself highlights gaps and discredits false assumptions. </p><p>Here we cover results from design projects that span several scales of complexity. First we describe the adaptation and experimental validation of protein functional assays on minute amounts of material. This work enables the application of cell-free protein expression tools in a high-throughput protein engineering pipeline, dramatically increasing turnaround time and reducing costs. The parts production pipeline can provide new building blocks for synthetic biology efforts with unprecedented speed. Tools to streamline the transition from the in vitro pipeline to conventional cloning were also developed. Next we detail an effort to expand the scope of a cysteine reactivity assay for generating information-rich datasets on protein stability and unfolding kinetics. We go on to demonstrate how the degree of site-specific local unfolding can also be determined by this method. This knowledge will be critical to understanding how proteins behave in the cellular context, particularly with regards to covalent modification reactions. Finally, we present results from an effort to engineer bacterial cell suicide in a population-dependent manner, and show how an underappreciated facet of plasmid physiology can produce complex oscillatory dynamics. This work is a prime example of engineering towards understanding.</p> en_US
dc.subject Biology, Molecular en_US
dc.subject Chemistry, Biochemistry en_US
dc.subject Engineering, Biomedical en_US
dc.subject gene circuits en_US
dc.subject local unfolding en_US
dc.subject protein engineering en_US
dc.subject protein stability en_US
dc.subject quantitative cysteine reactivity en_US
dc.subject synthetic biology en_US
dc.title Molecular Bioengineering: From Protein Stability to Population Suicide en_US
dc.type Dissertation en_US
dc.department Biochemistry en_US

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