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Title: Evaluation of different approaches to protein engineering and modulation
Keywords: Protein engineering, Intein, site-specific, immobilization, microarray, Sortase, evolution, phage display, 3CL protease
Issue Date: 4-May-2006
Citation: APARNA GIRISH (2006-05-04). Evaluation of different approaches to protein engineering and modulation. ScholarBank@NUS Repository.
Abstract: Three different approaches to protein engineering and modulation were evaluated. As the first approach to protein engineering, an intein mediated method to site-specifically modify proteins to allow immobilization onto the microarray was developed. Also as a complementary approach an intein mediated method that allows biotinylation of proteins produced from yeast host was developed. As a second approach, protein evolution of Sortase was considered. Sortase was displayed on the surface of the phage and subsequently shown to catalyze the transfer of biotinylated substrates onto itself. Thus when a library of random mutants of Sortase is displayed on the phage, only the active members will be selected by virtue of their biotin tag. Using this selection scheme one may evolve active members of Sortase from a pool of random mutants. Finally a random peptide library displayed on the phage was scanned for inhibitors to the SARS 3CL protease.
Appears in Collections:Master's Theses (Open)

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