Global ETD Search
Search theses and dissertations gathered from participating repositories worldwide. Every result links back to the library that holds it. No account is needed.
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Showing 1 to 16 of 16 for “"cell-free expression"”.
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Cell-free expression systems for the construction of artificial cells
Cell-free expression systems are widely used to synthesize proteins for subsequent further characterization, to manufacture potentially useful commercial end products, and to construct cellular mimics in the laboratory. The first part of the thesis explores the feasibility of preparing two of the …
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ToxIN-mediated resistance to and cell-free production of Salmonella bacteriophages
… of FelixO1 using an Escherichia coli-based cell-free expression system, demonstrating for the first time that a Salmonella phage can be produced in this manner. Taken together, this work illustrates that ToxIN may be of concern for food applications of phages while also establishing an …
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Lateral Transfer of DNA in Protocell-like Synthetic Cells
… how molecules could have moved between primitive cells is a central problem in Astrobiology and Geobiology. This thesis investigates whether electroporation can mediate the transfer of DNA into and between synthetic cells, potentially enabling gene expression in initially DNA-free compartments. …
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Resolving Signal Transduction in Complex Biological Environments
Cell surface receptors reside in complex biological environments along with their numerous extracellular ligands to transduce information across the membrane bilayer. These receptors regulate major functions including cell proliferation, facilitation of neuronal transmission, cell growth and …
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Technological advancements towards paper-based biomolecular diagnostics
… and translation, limiting their applicability to cellular contexts In vitro cell-free expression systems that contain transcription and translation machinery provide the environment necessary for biologically-based technologies to function independently of living cells. Our lab recently developed …
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Developing New Modalities for Biosensing using Synthetic Biology
… a mobile, functionalized microsurface with cell-free expression approaches. This biosensor demonstrated a dynamic range of 1- 100 ng/ml. In addition, I also explored the possibility of combining these biosensing systems with engineered living cells. By leveraging the tools of synthetic …
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Towards The Engineering Of A Synthetic Minimal Cell
Cell-free expression (CFE) systems are one of the main platforms for building a synthetic cell. A major drawback is the orthogonality of cell-free systems across species. To generate a CFE system compatible with recently established minimal cell constructs, we attempted to optimize a Mycoplasma …
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Engineering fluorescent imaging platforms to characterise molecular interactions with HIV-1 capsid
… measurements confirmed that binding of the host cell protein cyclophilin A (CypA) to the tubular lattice is substoichiometric due to steric hindrance between CypA molecules. Binding analysis of cleavage and polyadenylation specificity factor subunit 6 (CPSF6) expressed as a full-length protein in …
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Biochemical and biophysical investigations of N-linked glycosylation pathways in archaea
… unique polyprenyl-P-linked substrates. Lastly, a cell-free expression system was established for the efficient synthesis of Alg5, a yeast dolichyl-phosphate glucosyltransferase that shares high sequence similarity to AglK, the first glycosyltransferase in the M. voltae pathway. Dol-P-Glc was …
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Olfactory-related receptors : methods towards enabling structural and functional studies
… at low levels endogenously or in heterologous expression systems. Second, detergents capable of solubilizing and stabilizing these proteins in aqueous solution must be found. Third, the flexible nature of GPCRs can inhibit crystal lattice formation. Methods for overcoming each obstacle were …
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Expression and characterisation of metalloproteins from mycobacterium tuberculosis
… research presented in this thesis focuses on the expression and characterisation of targets from the Mycobacterium tuberculosis genome. Cell-free expression trials of 36 unique targets were performed. Initial screening resulted in soluble expression for 30 % of the targets and inclusion of …
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MOLECULAR CHARACTERIZATION AND EXPRESSION OF GENTISATE 1,2-DIOXYGENASE FROM PSEUDOMONAS ALCALIGENES NCIB 9867
… using three different heterologous E. coli overexpression systems, namely the 6X His tag, GST tag and the His-Patch ThioFusion™ systems as well as the in vitro cell free expression RTS 500 system. Binding of a 6X His-GOO fusion protein to the Ni2+ affinity column was poor when the 6X His tag was …
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A Bottom-Up Approach to Building an Artificial Cell by Self-Assembly
… now allow the formation of minimal or artificial cells composed of a minimum number of components, capable of performing specific functions. New developments contributing to the complexity of artificial cells or making their design more cell-like increases the number of possible applications. …
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Development of a saposin A based native-like phospholipid bilayer system for NMR studies
… II (pSRII), were tested. The parameters for expression and purification of ASR were first screened for the optimal yield. Although incorporation of ASR resulted in inhomogeneous particles due to imperfect experimental procedure, pSRII in SapA-DMPC nanoparticles showed high sample quality. The …