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dc.contributor.advisorEllington, Andrew D.en
dc.creatorDavidson, Eric Alanen
dc.date.accessioned2011-06-14T21:14:47Zen
dc.date.accessioned2011-06-14T21:15:17Zen
dc.date.available2011-06-14T21:14:47Zen
dc.date.available2011-06-14T21:15:17Zen
dc.date.issued2010-05en
dc.date.submittedMay 2010en
dc.identifier.urihttp://hdl.handle.net/2152/ETD-UT-2010-05-948en
dc.descriptiontexten
dc.description.abstractIn this work, we seek to expand synthetic in vitro biological systems by using water-in-oil emulsions to provide an environment conducive to directed evolution. We approach this primarily by utilizing a model transcription system, the T7 RNA polymerase and promoter, which is orthogonal to both bacterial and eukaryotic transcription systems and is highly functional in vitro. First, we develop a method to identify functional promoter sequences completely in vitro. This method is tested using the T7 RNA polymerase-promoter model system. We then configure the T7 transcription system as an ‘autogene’ and investigate how this positive feedback circuit (whereby a T7 promoter expresses a T7 RNA polymerase gene) functions across various in vitro platforms, including while compartmentalized. The T7 autogene can be envisioned as a self-replicating system when compartmentalized, and its use for directed evolution is examined. Finally, we look towards future uses for these in vitro systems. One interesting application is to expand the utilization of unnatural base pairs within the context of a synthetic system. We investigate the ability of T7 RNA polymerase to recognize and utilize unnatural base pairs within the T7 promoter, complementing existing work on the utilization of unnatural base pairs for in vitro replication and transcription with an investigation of more complex protein-dependent regulatory function. We envision this work as a foundation for future in vitro synthetic biology efforts.en
dc.format.mimetypeapplication/pdfen
dc.language.isoengen
dc.subjectSynthetic biologyen
dc.subjectCompartmentalized self-replicationen
dc.subjectT7 RNA polymeraseen
dc.subjectT7 autogeneen
dc.subjectIn vitro systemsen
dc.titleSynthetic transcription systemsen
dc.date.updated2011-06-14T21:15:17Zen
dc.contributor.committeeMemberBull, James J.en
dc.contributor.committeeMemberMarcotte, Edward M.en
dc.contributor.committeeMemberWhitman, Christian P.en
dc.contributor.committeeMemberYin, Yuhui W.en
dc.description.departmentCellular and Molecular Biologyen
dc.type.genrethesisen
thesis.degree.departmentCellular and Molecular Biologyen
thesis.degree.disciplineCellular and Molecular Biologyen
thesis.degree.grantorUniversity of Texas at Austinen
thesis.degree.levelDoctoralen
thesis.degree.nameDoctor of Philosophyen


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