A Combinatorial Approach for the Selection of Novel Bioactive Peptides

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dc.contributor.advisor Snell, R en
dc.contributor.author Lawrence, Shani en
dc.date.accessioned 2017-11-09T21:04:56Z en
dc.date.issued 2017 en
dc.identifier.uri http://hdl.handle.net/2292/36374 en
dc.description Full text is available to authenticated members of The University of Auckland only. en
dc.description.abstract Modern drug identification primarily focuses on the development and screening of prospective drug libraries in systems with known target properties. Although conceptually simple, new diseases and targets are being continuously discovered, and so fast efficient screening methods for increasingly larger compound libraries are required to identify prospective drugs. This is prohibitively expensive for smaller groups interested in rarer conditions, as high throughput screening requires advanced robotic equipment and large compound libraries which are not readily available to most academic labs. To overcome these economic and practical restrictions, the aim of the work presented here was the development of an inexpensive method for generating and screening novel bioactive compounds. Furthermore, the hope was that this method could be applied in any competent molecular biology lab. The basis for this method is the use of simple biological systems to screen complex peptide expression libraries developed using a molecular approach that explores the combinatorial search space of possible peptides. This proof of concept study developed and tested the methodology through combinatorial randomized sequence generation by degenerate primer PCR amplification, and screening this library in a modified toxin/antitoxin plasmid cloning system. For the first step, 30bp random sequences of DNA were synthesized as degenerate oligonucleotides and incorporated into two separate sites in the antitoxin coding sequence. Functional coding sequences were identified from the complex libraries by a simple live/dead screen after transformation in E.coli. This approach generated incredibly complex libraries. Screening and a relatively small scale analysis revealed multiple bioactive antitoxin sequences, which differed significantly from the known sequence. Although larger scale analysis, and further characterisation of the compounds is required, initial testing has indicated that this method is capable of generating very complex libraries simply and functions as an effective source for target specific, novel bioactive compounds. The next step for this proof of principle experiment is to undertake larger scale analysis of selected clones to determine codon patterns. Ultimately, the aim of the system is to select peptides for a human disease target. en
dc.publisher ResearchSpace@Auckland en
dc.relation.ispartof Masters Thesis - University of Auckland en
dc.relation.isreferencedby UoA99265060313702091 en
dc.rights Items in ResearchSpace are protected by copyright, with all rights reserved, unless otherwise indicated. Previously published items are made available in accordance with the copyright policy of the publisher. en
dc.rights Restricted Item. Available to authenticated members of The University of Auckland. en
dc.rights.uri https://researchspace.auckland.ac.nz/docs/uoa-docs/rights.htm en
dc.title A Combinatorial Approach for the Selection of Novel Bioactive Peptides en
dc.type Thesis en
thesis.degree.discipline Biomedical Science en
thesis.degree.grantor The University of Auckland en
thesis.degree.level Masters en
dc.rights.holder Copyright: The author en
pubs.elements-id 710641 en
pubs.record-created-at-source-date 2017-11-10 en

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