The Control and Detection of Biomolecules Using Functional DNA

Ali, Aysha (2019). The Control and Detection of Biomolecules Using Functional DNA. University of Birmingham. Ph.D.

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Abstract

Deoxyribonucleic acid (DNA) is the most important component of life and has many uses beyond the realm of natural biology. Synthetic oligonucleotide chemistry has allowed for the development of novel DNA based biotherapeutic and biosensing platforms that aim to overcome challenges faced within these fields. The versatility of synthetic DNA in the control and detection of biomolecules was investigated in this thesis using three topics:
1. The synthetic modification of the backbone of a thrombin binding aptamer (TBA). Thrombin is an important enzyme involved in the process of blood clotting and TBA competitively binds to thrombin at the same site as fibrinogen, disrupting the process of blood clotting. The TBA folds into a G-quadruplex formation and this structure is disrupted upon intramolecular photo-induced dimerisation of the two anthracene modifications. As a result, a switchable system was developed that allowed the reversible photo-control of thrombin activity using anthracene photochromism.
2. The synthetic modification, with one anthracene monomer, of the backbone of an oligonucleotide that probed for a Thymine (T) to Adenine (A) point variation in the BRAF gene, a known mutation in certain cancers. Genomic samples were amplified and successfully probed for the T or A point variant using fluorescence spectroscopy. The probe generated a linear dependence fluorescence plot, using synthetic DNA targets, to allow for the quantification of the allelic ratio of T to A within a sample of genomic DNA.
3. The bioconjugation of oligonucleotide probes to the M13-bacteriophage was utilised to design an assay, based on linear dichroism spectroscopy, for two potato virus strains. The assay was able to detect double stranded DNA targets at 0.03 nM, a level competitive with other detections methods that do not require amplification, and was further developed to detect two viral strains at once.

Type of Work:	Thesis (Doctorates > Ph.D.)
Award Type:	Doctorates > Ph.D.
Licence:	All rights reserved
College/Faculty:	Colleges (2008 onwards) > College of Engineering & Physical Sciences
School or Department:	School of Chemistry
Funders:	Engineering and Physical Sciences Research Council
Subjects:	Q Science > QD Chemistry
URI:	http://etheses.bham.ac.uk/id/eprint/9004

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