Praktiek, Jan (2022) Studying a Synthetic Phospholipid Biosynthesis Pathway Using a Cell Lysate Expression System. Master's Research Project 1, Biomolecular Sciences.
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Abstract
An important module in the construction of a bottom-up synthetic cell will be the generation of a cellular membrane, which will likely be composed of phospholipids. Previous research has indicated the possibility of generating a functional cellular membrane using a synthetic phospholipid biosynthesis pathway. This biosynthesis pathway was already shown to be functional in an expression system using PURE. The aim of this research project was to study the phospholipid biosynthesis pathway in a cell lysate expression system. The influence of insertases SecYEG and YidC on co-translational insertion of this biosynthesis pathway into liposomal membranes was studied using mass spectrometry. Furthermore, the possibility of bulk lipid production in such a lysate expression system was elucidated using thin layer chromatography as an analysis method. It was found that while such a lysate expression system does allow for robust production of lipids, it is also prone to interference by background processes. However, analysis using thin layer chromatography coupled with fluorescent imaging also seemed to indicate the potential for bulk lipid production using such a system.
| Item Type: | Thesis (Master's Research Project 1) |
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| Supervisor name: | Driessen, A.J.M. and Uijl, M.J. den |
| Degree programme: | Biomolecular Sciences |
| Thesis type: | Master's Research Project 1 |
| Language: | English |
| Date Deposited: | 03 Oct 2022 11:35 |
| Last Modified: | 03 Oct 2022 11:35 |
| URI: | https://fse.studenttheses.ub.rug.nl/id/eprint/28799 |
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