Engineering leave-one-out GFP based biosensors
Authors
Fraser, Keith
Issue Date
2016-08
Type
Electronic thesis
Thesis
Thesis
Language
ENG
Keywords
Biology
Alternative Title
Abstract
To this end we proposed the development of a biosensor platform based on the green fluorescent protein (GFP) called leave-one-out GFP (LOO-GFP) biosensors. This platform is built on the principle that when one of the 11 β-strands in GFP is excised (LOO-GFP) there is a loss in the native fluorescence of the molecule that can be reconstituted upon the addition of the missing peptide. Additionally, the pocket created as a result of the omission of any of the β-strands can be redesigned to accommodate an exogenous peptide sequence, allowing for LOO-GFPs to function as designable biosensors. Since this approach is a redesign of the existing molecule, we conducted a mechanistic study of the structure function relationship in GFP so as to identify the best engineering strategies. This work focuses on how the production of the fluorescent para-hydroxbenzylidene imidazolinone chromophore is regulated, the identification of optimal design scaffolds and developing methods that can be used to design LOO-GFP biosensors for diagnostics and environment surveillance.
Description
August 2016
School of Science
School of Science
Full Citation
Publisher
Rensselaer Polytechnic Institute, Troy, NY