Engineering leave-one-out GFP based biosensors

Authors
Fraser, Keith
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Other Contributors
Bystroff, Christopher, 1960-
Dordick, Jonathan S.
Gilbert, Susan P.
Wang, Chunyu
Lakshmi, K. V.
Issue Date
2016-08
Keywords
Biology
Degree
PhD
Terms of Use
This electronic version is a licensed copy owned by Rensselaer Polytechnic Institute, Troy, NY. Copyright of original work retained by author.
Full Citation
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
Department
Dept. of Biological Sciences
Publisher
Rensselaer Polytechnic Institute, Troy, NY
Relationships
Rensselaer Theses and Dissertations Online Collection
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