Dokument: Engineering and implementation of synthetic (opto-) genetic tools for Ustilago maydis and reconstruction of (signaling) pathways in orthogonal Systems
| Titel: | Engineering and implementation of synthetic (opto-) genetic tools for Ustilago maydis and reconstruction of (signaling) pathways in orthogonal Systems | |||||||
| URL für Lesezeichen: | https://docserv.uni-duesseldorf.de/servlets/DocumentServlet?id=55387 | |||||||
| URN (NBN): | urn:nbn:de:hbz:061-20220302-093938-2 | |||||||
| Kollektion: | Dissertationen | |||||||
| Sprache: | Englisch | |||||||
| Dokumententyp: | Wissenschaftliche Abschlussarbeiten » Dissertation | |||||||
| Medientyp: | Text | |||||||
| Autor: | M. Sc. Heucken, Nicole [Autor] | |||||||
| Dateien: |
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| Beitragende: | Prof. Dr. Zurbriggen, Matias [Gutachter] Prof. Dr. Feldbrügge, Michael [Gutachter] | |||||||
| Dewey Dezimal-Klassifikation: | 500 Naturwissenschaften und Mathematik » 570 Biowissenschaften; Biologie | |||||||
| Beschreibung: | Production of pharmaceutically relevant substances is traditionally realized by
chemical synthesis with several drawbacks such as limited product diversity, accumulation of toxic waste and low outcome despite high costs. For these reasons, more and more compounds are produced biotechnologically and new production platforms are constantly needed to be able to satisfy the increasing demands of society. The dimorphic basidiomycete fungus Ustilago maydis is used as a model organism for many biological processes, and more recently, a biotechnological role, as a production platform for high value compounds, is emerging. During this work, several basic and advanced tools for the pathway engineering in U. maydis were constructed and tested. The functionality of the quantitative luminescent reporters FLuc, RLuc and GLuc was proven. With their help, a fast screening platform for the efficient generation of U. maydis strains was established. Additionally, their use as a normalization element was demonstrated in an inductionbased gene expression quantification system. They were further applied for the testing and characterization of several synthetic tools, such as IRES sequences and bidirectional promoters, but also DNA-binding protein – DNA operating sequence interactions, and chemical as well as light regulated gene expression. While IRES sequences and chemical and UV-B-light regulated gene expression are, at this stage, not yet functional, two bidirectional promoters, several orthogonal unidirectional promoters and the PIP-PIR3 system, as the base for split transcription factor systems, have shown to be functional and useful in U. maydis. In the second part of this work, certain aspects of phytohormone signalling have been analysed in more detail. Fluorescence microscopy studies have proven the interaction of the DELLA proteins RGA and GAI with the COP1/SPA1 complex, and shown that mainly SPA1 is responsible for this interaction. Moreover, the functionality of induction-based, ratiometric, luminescent phytohormone biosensors was demonstrated on the example of the abscisic acid receptor PYL8, which is upregulated upon hormone perception. Efficiency of this sensor was increased by downregulation of the receptor protein level by coexpression of the E3 ubiquitin ligase RFA4 before hormone treatment. Lastly, a potential-induction-fold-determination-vector was constructed using an updated version of AQUA cloning, with which dynamic ranges of phytohormone biosensors can be estimated in several organisms. | |||||||
| Lizenz: |  Urheberrechtsschutz | |||||||
| Fachbereich / Einrichtung: | Mathematisch- Naturwissenschaftliche Fakultät | |||||||
| Dokument erstellt am: | 02.03.2022 | |||||||
| Dateien geändert am: | 02.03.2022 | |||||||
| Promotionsantrag am: | 16.06.2020 | |||||||
| Datum der Promotion: | 15.12.2020 |
