Generation of Pseudomonas mutants for investigating a model mixed species biofilm

Mixed-species microbial biofilms are the predominant form of biofilms in nature, however, majority of studies on biofilms have focused on single-species biofilms. Unique interspecies interactions could shape the biofilm community structure and functions in a manner that is not represented in single-...

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Main Author: Neo, Desmond
Other Authors: Scott Rice
Format: Final Year Project
Language:English
Published: 2018
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Online Access:http://hdl.handle.net/10356/74624
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Institution: Nanyang Technological University
Language: English
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spelling sg-ntu-dr.10356-746242023-02-28T18:01:43Z Generation of Pseudomonas mutants for investigating a model mixed species biofilm Neo, Desmond Scott Rice School of Biological Sciences DRNTU::Science Mixed-species microbial biofilms are the predominant form of biofilms in nature, however, majority of studies on biofilms have focused on single-species biofilms. Unique interspecies interactions could shape the biofilm community structure and functions in a manner that is not represented in single-species biofilms. Thus, a model mixed-species biofilm comprised of Klebsiella pneumoniae (KP-1), Pseudomonas aeruginosa (PAO1) and Pseudomonas protegens (Pf-5) was selected to investigate the differentially expressed genes between single-species and mixed-species biofilms to identify key genes within Pf-5 involved in overall biofilm structure via mutagenesis studies. This study involved the fluorescent tagging of PAO1 via specific insertion mutagenesis involving a Tn7 transposon. Deletion mutagenesis of differentially expressed genes in Pf-5 were performed, with PFL_5251 as the target gene to develop a reliable method for deletion mutagenesis in Pf-5 where multiple key methods were identified at each stage. Both electroporation and conjugation were evaluated for the introduction of pK18 assembled with deletion construct into Pf-5. Both failed to select for transformants and transconjugants until the antibiotic concentrations were optimized. PFL_5251 mutants were generated and confirmed for Pf-5 via sequence alignment. This paved the way for further analysis with bioimaging techniques to help improve understanding and determination of genes important in mixed-species biofilms. Bachelor of Science in Biological Sciences 2018-05-22T06:49:09Z 2018-05-22T06:49:09Z 2018 Final Year Project (FYP) http://hdl.handle.net/10356/74624 en Nanyang Technological University 49 p. application/pdf
institution Nanyang Technological University
building NTU Library
continent Asia
country Singapore
Singapore
content_provider NTU Library
collection DR-NTU
language English
topic DRNTU::Science
spellingShingle DRNTU::Science
Neo, Desmond
Generation of Pseudomonas mutants for investigating a model mixed species biofilm
description Mixed-species microbial biofilms are the predominant form of biofilms in nature, however, majority of studies on biofilms have focused on single-species biofilms. Unique interspecies interactions could shape the biofilm community structure and functions in a manner that is not represented in single-species biofilms. Thus, a model mixed-species biofilm comprised of Klebsiella pneumoniae (KP-1), Pseudomonas aeruginosa (PAO1) and Pseudomonas protegens (Pf-5) was selected to investigate the differentially expressed genes between single-species and mixed-species biofilms to identify key genes within Pf-5 involved in overall biofilm structure via mutagenesis studies. This study involved the fluorescent tagging of PAO1 via specific insertion mutagenesis involving a Tn7 transposon. Deletion mutagenesis of differentially expressed genes in Pf-5 were performed, with PFL_5251 as the target gene to develop a reliable method for deletion mutagenesis in Pf-5 where multiple key methods were identified at each stage. Both electroporation and conjugation were evaluated for the introduction of pK18 assembled with deletion construct into Pf-5. Both failed to select for transformants and transconjugants until the antibiotic concentrations were optimized. PFL_5251 mutants were generated and confirmed for Pf-5 via sequence alignment. This paved the way for further analysis with bioimaging techniques to help improve understanding and determination of genes important in mixed-species biofilms.
author2 Scott Rice
author_facet Scott Rice
Neo, Desmond
format Final Year Project
author Neo, Desmond
author_sort Neo, Desmond
title Generation of Pseudomonas mutants for investigating a model mixed species biofilm
title_short Generation of Pseudomonas mutants for investigating a model mixed species biofilm
title_full Generation of Pseudomonas mutants for investigating a model mixed species biofilm
title_fullStr Generation of Pseudomonas mutants for investigating a model mixed species biofilm
title_full_unstemmed Generation of Pseudomonas mutants for investigating a model mixed species biofilm
title_sort generation of pseudomonas mutants for investigating a model mixed species biofilm
publishDate 2018
url http://hdl.handle.net/10356/74624
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