- Page 1 and 2: Characterisation of pathogenicity i
- Page 3: Statement of Originality This accom
- Page 7 and 8: Table of contents 1 INTRODUCTION 1
- Page 9 and 10: 4 THE CONTRIBUTION OF PAPI-1 AND PA
- Page 11 and 12: 1 Introduction 1 INTRODUCTION 1 1.1
- Page 13 and 14: Cell-surface Proteases Haemolysins
- Page 15 and 16: 1.2.1 Pathogenicity islands: a subg
- Page 17 and 18: appear to be related and there have
- Page 19 and 20: assessed in number of assays and mo
- Page 21 and 22: TAR cloning is a method designed by
- Page 23 and 24: Figure 1-3 Top plate shows colony g
- Page 25 and 26: Figure 1-5 depicts the principles o
- Page 27 and 28: Balb/c mice. The models described b
- Page 29 and 30: urns-sepsis model, but not in the n
- Page 31 and 32: ORF ID Gene name Gene function PA14
- Page 33 and 34: functional activities it could be s
- Page 35 and 36: Figure 1-6 Schematic showing the OR
- Page 37 and 38: RhlI Autoinducer RhlR Rhl Regulon G
- Page 39 and 40: 1.5.2 Pseudomonas aeruginosa LES Th
- Page 41 and 42: 2 Material and Methods 2 MATERIAL A
- Page 43 and 44: 2.1.2 Strains used Pseudomonas aeru
- Page 45 and 46: Plasmids Plasmid Description Refere
- Page 47 and 48: In the case of capture vector const
- Page 49 and 50: Primer name Sequence Capture vector
- Page 51 and 52: The relevant homology sequence was
- Page 53 and 54: Important to the use of the capture
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2.3.4 Electroporation: Transfer of
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2.4.2 Infection dose preparation Th
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mould was then added to the metal c
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3 Analysis of genomic islands captu
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3.1 Capture experiments involving g
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DNA No of colonies Control No DNA 0
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were designed based on the assumpti
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3.2.1 KR115-lys10 and KR159-lys10 A
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estriction digestion with I-SceI to
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generated. The most significant nuc
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they both produced three fragments
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elated genomic islands and this res
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All the Yersinia species available
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Leicester). E105-leuX was the first
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The strains highlighted with alignm
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predict this protein to be a dnaJ-c
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Figure 3-15 depicts the codon usage
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ORF Gene range Blastx Description y
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ecognition sequences are found with
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(S: 150, E: 1e-37). This predicts t
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having multiple copies of itself. A
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The capture vectors used during thi
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cloning and their findings was inco
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The genomic island capture method c
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unpublished). This was achieved by
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Pseudomonas aeruginosa PA14 (UCBPP-
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stressful’ to the bacteria and th
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log CFU/mg 4 3 2 1 0 0 2 4 6 8 Days
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mice exhibited visible symptoms at
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4.4.1 Survival times post-infection
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(P
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4.4.4 Intravenous infection A quest
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Symptom score Symptom score Symptom
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Fold increase Fold increase 15 10 5
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Figure 4-15 Histopathology lung sec
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4.4.7 Progression of disease post-i
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Figure 4-19 Leukocytes numbers moni
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interesting result is that the ∆P
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Unfortunately, 65% of the ORFs with
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morphology. ∆PAPI-1∆PAPI-2 leav
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The intravenous sepsis model data s
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in PAO1(Lee et al. 2006). They show
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shown that deletion of the exoU gen
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5 The role of quorum-sensing in the
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Figure 5-1 shows overnight growth o
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Quorum-sensing is bacterial cell de
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Interestingly, the quorum-sensing d
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Symptom score Symptom score Symptom
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LES431 LESB58 LESB65 LES400 � �
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Additional comparisons of the CFU r
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log CFU/mg log CFU/mg log CFU/ml 5
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For LESB58-infected mice the histop
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Strain Mouse Timepoint HB CI GC Sco
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Score: 4 HB Score: 4 Score: 4 B HCI
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post-infection to that of healthy m
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Tang et al. (1996) performed additi
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Figure 5-14 PFGE comparison of the
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isolates have a similar virulence i
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The data presented in this thesis s
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than PAO1. Despite the bacterial lu
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6 Thesis conclusion The underlying
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Hopefully the methodologies describ
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1 2 3 4 5 6 7 8 9 10 11 12 13 λHin
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AQ: 1 AQ: 2 AQ: 3 AQ: 4 AQ: 5 AQ: 6
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AQ: 18 AQ: 19 AQ: 20 F1 AQ: 21 tapr
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AQ: 32 AQ: 33 AQ: 34 tapraid5/z9d-j
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AQ: 40 AQ: 41 DISCUSSION tapraid5/z
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AQ: 48 tapraid5/z9d-jid/z9d-jid/z9d
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tapraid5/z9d-jid/z9d-jid/z9d01009/z
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8 Bibliography AL-ALOUL, M., CRAWLE
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D'ARGENIO, D.A., WU, M., HOFFMAN, L
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genomic islands in Pseudomonas aeru
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MURRAY, A.W. and SZOSTAK, J.W., 198
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specificity of chaperone-usher mach
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VAN HEECKEREN, A.M. and SCHLUCHTER,