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Abstract: DNA sequencing of phage display products presents a combination of two technologies capable of providing a high throughput means of screening protein-protein and protein-substrate interactions. Phage display presents a system for the controlled interrogation of molecular interactions at the expressed protein level, yet uses the encoding DNA as the analyte. Recent advances in DNA sequenci... read moreng technologies have rendered this technology a tractable readout for a variety of applications, including phage display. The method described circumvents the need for any infection of phagemid into bacteria with the intent of removing amplification as a source of bias. In order to understand the utility of this technology for screening and discovery of amino acid motifs involved in extracellular matrix adhesion, highly diverse phage libraries were screened against silk, collagen, and polystyrene substrates. Products were PCR amplified and prepared for Illumina sequencing. Recovered sequences were translated in silico to peptide sequences and aligned to known extracellular matrix protein sequences to identify regions of substrate interaction.
Thesis (M.S.)--Tufts University, 2012.
Submitted to the Dept. of Biomedical Engineering.
Advisors: David Kaplan, and Nathaniel Cosper.
Committee: Hyumin Yi, and Qiaobing Xu.
Keyword: Biomedical engineering.read less
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