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Identifier 000399744
Title Natural fibrous proteins as scaffolds for nanotechnology
Alternative Title Φυσικές ινώδεις πρωτεΐνες ως ικριώματα νανοτεχνολογίας
Author Γεωργιλής, Ευάγγελος Μ.
Thesis advisor Κοκκινίδης, Μιχαήλ
Reviewer Μητράκη, Άννα
Πετράτος, Κυριάκος
Abstract The present study was conducted in the laboratory of Natural Biomaterials, Dept. of Materials Science and Technology, University of Crete. The research was focused on the development of self-assembled protein materials that can be used as scaffolds in nanotechnology. The system used for this purpose was the â-helical domain of the P22 phage tailspike (Bhx). The sequence of Bhx is aggregation-prone and thermoresponsive. Moreover, it binds to oligosaccharides and has been shown to form irreversible, fibrous aggregates during refolding experiments. In this study, spherical and fibrous microstructures of Bhx were developed and observed via electron microscopy. These studies revealed a hierarchical formation of the microstructures from nano-filaments. Inspired by the â-helical virulence factor of Bordetella pertussis that mediates mammalian cell adhesion, an RGD-motif was inserted to the sequence of Bhx via site-directed mutagenesis for the development of biocompatible fibrous scaffolds that might promote cell adhesion and proliferation. Furthermore, an emerald green fluorescent protein (GFPem) and a biotinylation peptide sequence were introduced to the sequence of Bhx via the seamless cloning method. GFPem was fused to both the N- and C-terminus of Bhx, providing data on its degradation resistance and functionality of enzymes in inclusion bodies.
Language English
Issue date 2016-03-18
Collection   School/Department--School of Sciences and Engineering--Department of Biology--Post-graduate theses
  Type of Work--Post-graduate theses
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