Host defence peptide (HDP) human beta defensin 9 (HBD9)

Introduction: The emergence of antibiotic resistance has led to the continuing search for discovery of effective antibiotics. Host defence peptides (HDPs) confer defence mechanisms against infection and investigation of their specific roles and interplays are ongoing. Among the HDPs, defensins are...

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Main Author: Omar, Nazri
Format: Thesis (University of Nottingham only)
Language:English
Published: 2016
Subjects:
Online Access:https://eprints.nottingham.ac.uk/33280/
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author Omar, Nazri
author_facet Omar, Nazri
author_sort Omar, Nazri
building Nottingham Research Data Repository
collection Online Access
description Introduction: The emergence of antibiotic resistance has led to the continuing search for discovery of effective antibiotics. Host defence peptides (HDPs) confer defence mechanisms against infection and investigation of their specific roles and interplays are ongoing. Among the HDPs, defensins are a group of effector molecules which plays important roles in humans. Although several stereotypes of human beta defensins (hBDs) such as the hBD1-3 are well studied, other members including the human beta defensin 9 (hBD9), are not entirely known. Understanding the properties of these HDPs will enable us to discover a safe and efficacious, broad-spectrum and resistance-free antibiotic for therapeutic application in the future. Purpose: The purpose of this study is to clone the DEFB109 gene, express and purify the hBD9 propeptide, before determining the hBD9 propeptide antimicrobial property using a recombinant system in Escherichia coli. Methods: The second exon of the DEFB109 was amplified through reverse transcription polymerase chain reaction (RT-PCR) and inserted into selected plasmid vectors. The recombinant plasmid construct was cloned, and transformed into E coli expression host. The correctly transformed colonies were selected before the plasmid constructs were purified and verified through nucleotide sequencing. Expression and purification of the hBD9 propeptide were carried out and antimicrobial property of the peptide was investigated. Result: HBD9 fusion protein was successfully expressed and purified. It was shown to have antimicrobial efficacy against Staphylococcus aureus and Pseudomonas aeruginosa. The effect of the free hBD9 propeptide against wider spectrum of organisms needs to be studied in the future.
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format Thesis (University of Nottingham only)
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language English
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spelling nottingham-332802025-02-28T13:27:09Z https://eprints.nottingham.ac.uk/33280/ Host defence peptide (HDP) human beta defensin 9 (HBD9) Omar, Nazri Introduction: The emergence of antibiotic resistance has led to the continuing search for discovery of effective antibiotics. Host defence peptides (HDPs) confer defence mechanisms against infection and investigation of their specific roles and interplays are ongoing. Among the HDPs, defensins are a group of effector molecules which plays important roles in humans. Although several stereotypes of human beta defensins (hBDs) such as the hBD1-3 are well studied, other members including the human beta defensin 9 (hBD9), are not entirely known. Understanding the properties of these HDPs will enable us to discover a safe and efficacious, broad-spectrum and resistance-free antibiotic for therapeutic application in the future. Purpose: The purpose of this study is to clone the DEFB109 gene, express and purify the hBD9 propeptide, before determining the hBD9 propeptide antimicrobial property using a recombinant system in Escherichia coli. Methods: The second exon of the DEFB109 was amplified through reverse transcription polymerase chain reaction (RT-PCR) and inserted into selected plasmid vectors. The recombinant plasmid construct was cloned, and transformed into E coli expression host. The correctly transformed colonies were selected before the plasmid constructs were purified and verified through nucleotide sequencing. Expression and purification of the hBD9 propeptide were carried out and antimicrobial property of the peptide was investigated. Result: HBD9 fusion protein was successfully expressed and purified. It was shown to have antimicrobial efficacy against Staphylococcus aureus and Pseudomonas aeruginosa. The effect of the free hBD9 propeptide against wider spectrum of organisms needs to be studied in the future. 2016-07-19 Thesis (University of Nottingham only) NonPeerReviewed application/pdf en arr https://eprints.nottingham.ac.uk/33280/1/Nazri%20thesis%20final%20revised%20and%20checked%20corrections%20V3%20170516%20amended.pdf Omar, Nazri (2016) Host defence peptide (HDP) human beta defensin 9 (HBD9). PhD thesis, University of Nottingham. Antimicrobial peptide Defensin Beta defensin Human beta defensin Human defence peptide
spellingShingle Antimicrobial peptide
Defensin
Beta defensin
Human beta defensin
Human defence peptide
Omar, Nazri
Host defence peptide (HDP) human beta defensin 9 (HBD9)
title Host defence peptide (HDP) human beta defensin 9 (HBD9)
title_full Host defence peptide (HDP) human beta defensin 9 (HBD9)
title_fullStr Host defence peptide (HDP) human beta defensin 9 (HBD9)
title_full_unstemmed Host defence peptide (HDP) human beta defensin 9 (HBD9)
title_short Host defence peptide (HDP) human beta defensin 9 (HBD9)
title_sort host defence peptide (hdp) human beta defensin 9 (hbd9)
topic Antimicrobial peptide
Defensin
Beta defensin
Human beta defensin
Human defence peptide
url https://eprints.nottingham.ac.uk/33280/