Title page for ETD etd-11082006-161405


Document Type Doctoral Thesis
Author Steyn, Bridgitta
URN etd-11082006-161405
Document Title Proteomic analysis of the biofilm and biofilm-associated phenotypes of Pseudomonas aeruginosa cultured in batch
Degree PhD (Microbiology)
Department Microbiology and Plant Pathology
Supervisor
Advisor Name Title
Prof J Theron Committee Chair
Prof V S Brozel Committee Co-Chair
Keywords
  • Pseudomonas aeruginosa
  • biofilm and biofilm-associated phenotypes
  • biofilm-forming organisms
Date 2006-05-02
Availability unrestricted
Abstract
Pseudomonas aeruginosa is one of the most studied biofilm-forming organisms and has emerged as a model organism in the study of surface- and biofilm-induced gene expression. The transition from a planktonic to a biofilm mode of growth results in diverse changes in gene expression, which causes the attaching cells to become phenotypically and metabolically distinct from their planktonic counterparts. In this study, a proteomic approach was used to study differences in protein profiles obtained from 18-h old P. aeruginosa PAO1 (DSM 1707) planktonic, surface influenced planktonic (SIP) and biofilm populations grown in batch in the absence or presence of a glass wool substratum. Glass wool as an attachment substratum not only supported growth of biofilms, but it also allowed for the separation of the biofilm biomass from the surrounding surface influenced planktonic (SIP) cells for further characterisation. Comparative analysis of the respective proteomes indicated striking differences in the protein patterns of planktonic, biofilm and SIP cells and several uniquely expressed proteins were seen on the 2-DE protein maps of the respective populations. Whereas a general down-regulation of protein expression was seen in the biofilm cells, in SIP cells, expression of the proteins was generally up-regulated. The results confirmed that the biofilm population differs from the planktonic population and indicated that the SIP population is not merely a mixture of planktonic and biofilm cells but rather a unique phenotype.

Several differentially expressed protein spots were selected and identified using a combination of N-terminal protein sequencing and peptide mass fingerprinting. The proteins comprised mostly of outer membrane or membrane-associated proteins. Based on these analyses, a mutant P. aeruginosa strain, deficient in outer membrane protein OprG, was generated and its ability to form biofilms on a glass wool substratum was compared with that of the wild-type P. aeruginosa strain. The mutant strain was attachment-proficient but biofilm-deficient, suggesting that OprG plays a role in P. aeruginosa biofilm development under the culturing conditions used in this study.

2005, University of Pretoria. All rights reserved. The copyright in this work vests in the University of Pretoria. No part of this work may be reproduced or transmitted in any form or by any means, without the prior written permission of the University of Pretoria.

Please cite as follows:

Steyn, B 2005, Proteomic analysis of the biofilm and biofilm-associated phenotypes of Pseudomonas aeruginosa cultured in batch, PhD thesis, University of Pretoria, Pretoria, viewed yymmdd < http://upetd.up.ac.za/thesis/available/etd-11082006-161405/ >

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