A self-actuated cellular protein delivery machinery

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buir.advisorŞeker, Urartu Özgür Şafak
dc.contributor.authorAhan, Recep Erdem
dc.date.accessioned2018-08-10T13:32:23Z
dc.date.available2018-08-10T13:32:23Z
dc.date.copyright2018-08
dc.date.issued2018-08
dc.date.submitted2018-08-08
dc.departmentGraduate Program in Materials Science and Nanotechnologyen_US
dc.descriptionCataloged from PDF version of article.en_US
dc.descriptionThesis (M.S.): Bilkent University, Department of Materials Science and Nanotechnology, İhsan Doğramacı Bilkent University, 2018.en_US
dc.descriptionIncludes bibliographical references (leaves 86-98).en_US
dc.description.abstractOwing to increase the knowledge on biology and available tools for genetic manipulations, biological systems are engineered to perform complex tasks. They can be designed to degrade toxic molecules in environment, produce and deliver complex biological drugs, or process and synthesize valuable materials. Hence, the cellular machines hold great promises to solve world problems such as global warming, world hunger, cancer and so forth. However, most of the complex tasks require protein release to extracellular space in a controlled manner. Development of efficient cellular machines is hampered by lack of convenient strategy for controlled protein release as many of proposed secretion systems are limited in a narrow focused application. In this thesis, we are proposing a novel bifunctional self-exciting protein delivery system for broader applications. The proposed protein delivery machine harbours a genetic circuit that is able to display protein-of-interest on cell surface and to secrete to extracellular space in case of need. To do so, we engineered the autotransporter protein Ag43 to display POI with TEV protease recognition site on the cell surface of Escherichia coli (E. coli). The release of displayed POI was achieved and systematically optimized in vitro via addition of purified TEV protease. To accomplish the self-exciting and controlled release of POI by cells, TEV protease was aimed to be expressed and translocated to extracellular space to cleave the recognition site between POI and Ag43 protein. Four different secretion strategies was employed to secrete TEV protease to extracellular space. While cleavage of POI from cell surface can’t be accomplished through secretion of TEV protease by type I system, YebF fusion, and co-expressing lysis gene, codisplaying TEV protease on the cell surface can release the POI. Our data revealed that release of POI can be tuned with controlling the amount of TEV protease on the cell surface. Considering the simplicity of protein display as well as ability to express Ag43 protein in various organisms, the proposed system can be implemented in more complex genetic circuits and used in diverse applications.en_US
dc.description.degreeM.S.en_US
dc.description.statementofresponsibilityby Recep Erdem Ahan.en_US
dc.embargo.release2021-08-08
dc.format.extentxxii, 129 leaves : illustrations (some color) : charts ; 30 cm.en_US
dc.identifier.itemidB158766
dc.identifier.urihttp://hdl.handle.net/11693/47738
dc.language.isoEnglishen_US
dc.publisherBilkent Universityen_US
dc.rightsinfo:eu-repo/semantics/openAccessen_US
dc.subjectProtein Secretionen_US
dc.subjectCellular Machinesen_US
dc.subjectAutotransporter Proteinen_US
dc.subjectProtein Displayen_US
dc.subjectTEV Proteaseen_US
dc.titleA self-actuated cellular protein delivery machineryen_US
dc.title.alternativeKendiliğinden harekete geçebilen hücresel protein taşıma aracıen_US
dc.typeThesisen_US

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