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Biochemistry, Biophysics, and Structural Biology Commons

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Molecular Biology

Series

University of Connecticut

Honors Scholar Theses

2017

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Full-Text Articles in Biochemistry, Biophysics, and Structural Biology

Engineering A Fluorescent Protease Sensor For In Vivo Protein Detection, Thomas C. Kinard Jan 2017

Engineering A Fluorescent Protease Sensor For In Vivo Protein Detection, Thomas C. Kinard

Honors Scholar Theses

This report details the results of an ongoing project to engineer a mutant form of Red Fluorescent Protein (RFP) variant mCherry that acts as a real-time in vivo protease sensor. The sought-after mutant only becomes fluorescent when exposed to Tobacco Etch Virus (TEV) Protease, this system’s model protease. This will be accomplished via the insertion of the TEV Protease Recognition Site (TEV-PRS) in such a position that, before cleavage, will prevent the protein from folding to fluorescent conformation, but upon cleavage, will allow for fluorescent conformation to occur. The cylindrical structure of the protein, composed of beta-pleated sheets, contains ...