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D'ARGENIO, D.A., WU, M., HOFFMAN, L.R., KULASEKARA, H.D., DEZIEL, E., SMITH, E.E., NGUYEN, H., ERNST, R.K., LARSON FREEMAN, T.J., SPENCER, D.H., BRITTNACHER, M., HAYDEN, H.S., SELGRADE, S., KLAUSEN, M., GOODLETT, D.R., BURNS, J.L., RAMSEY, B.W. and MILLER, S.I., 2007. Growth phenotypes <strong>of</strong> <strong>Pseudomonas</strong> aerug<strong>in</strong>osa lasR mutants adapted to <strong>the</strong> airways <strong>of</strong> cystic fibrosis patients. Molecular microbiology, 64(2), pp. 512-533. DRENKARD, E. and AUSUBEL, F.M., 2002. <strong>Pseudomonas</strong> bi<strong>of</strong>ilm formation and antibiotic resistance are l<strong>in</strong>ked to phenotypic variation. Nature, 416(6882), pp. 740- 743. DUAN, K. and SURETTE, M.G., 2007. Environmental regulation <strong>of</strong> <strong>Pseudomonas</strong> aerug<strong>in</strong>osa PAO1 Las and Rhl <strong>quorum</strong>-<strong>sens<strong>in</strong>g</strong> systems. Journal <strong>of</strong> Bacteriology, 189(13), pp. 4827-4836. ERMOLAEVA, M.D., 2001. Synonymous codon usage <strong>in</strong> bacteria. Current Issues <strong>in</strong> Molecular Biology, 3(4), pp. 91-97. FINCK-BARBANCON, V., GORANSON, J., ZHU, L., SAWA, T., WIENER- KRONISH, J.P., FLEISZIG, S.M., WU, C., MENDE-MUELLER, L. and FRANK, D.W., 1997. ExoU expression by <strong>Pseudomonas</strong> aerug<strong>in</strong>osa correlates with acute cytotoxicity and epi<strong>the</strong>lial <strong>in</strong>jury. Molecular microbiology, 25(3), pp. 547-557. FINCK-BARBANCON, V., GORANSON, J., ZHU, L., SAWA, T., WIENER- KRONISH, J.P., FLEISZIG, S.M., WU, C., MENDE-MUELLER, L. and FRANK, D.W., 1997. ExoU expression by <strong>Pseudomonas</strong> aerug<strong>in</strong>osa correlates with acute cytotoxicity and epi<strong>the</strong>lial <strong>in</strong>jury. Molecular microbiology, 25(3), pp. 547-557. FOTHERGILL, J.L., PANAGEA, S., HART, C.A., WALSHAW, M.J., PITT, T.L. and WINSTANLEY, C., 2007. Widespread pyocyan<strong>in</strong> over-production among isolates <strong>of</strong> a cystic fibrosis epidemic stra<strong>in</strong>. BMC microbiology, 7, pp. 45. FUHRMANN, M., HAUSHERR, A., FERBITZ, L., SCHODL, T., HEITZER, M. and HEGEMANN, P., 2004. Monitor<strong>in</strong>g dynamic expression <strong>of</strong> nuclear genes <strong>in</strong> Chlamydomonas re<strong>in</strong>hardtii by us<strong>in</strong>g a syn<strong>the</strong>tic luciferase reporter gene. Plant Molecular Biology, 55(6), pp. 869-881. FURUKAWA, S., KUCHMA, S.L. and O'TOOLE, G.A., 2006. Keep<strong>in</strong>g <strong>the</strong>ir options open: acute versus persistent <strong>in</strong>fections. Journal <strong>of</strong> Bacteriology, 188(4), pp. 1211-1217. GARAU, J. and GOMEZ, L., 2003. <strong>Pseudomonas</strong> aerug<strong>in</strong>osa pneumonia. Current op<strong>in</strong>ion <strong>in</strong> <strong>in</strong>fectious diseases, 16(2), pp. 135-143. GIETZ, R.D. and WOODS, R.A., 2001. Genetic transformation <strong>of</strong> yeast. Biotechniques, 30(4), pp. 816-20, 822-6, 828 passim. GIETZ, R.D. and WOODS, R.A., 2002. Transformation <strong>of</strong> yeast by lithium acetate/s<strong>in</strong>gle-stranded carrier DNA/polyethylene glycol method. Methods <strong>in</strong> enzymology, 350, pp. 87-96. 173
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Characterisation of pathogenicity i
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Statement of Originality This accom
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% Percent Abbreviations ºC Degrees
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Table of contents 1 INTRODUCTION 1
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4 THE CONTRIBUTION OF PAPI-1 AND PA
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1 Introduction 1 INTRODUCTION 1 1.1
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Cell-surface Proteases Haemolysins
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1.2.1 Pathogenicity islands: a subg
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appear to be related and there have
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assessed in number of assays and mo
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TAR cloning is a method designed by
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Figure 1-3 Top plate shows colony g
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Figure 1-5 depicts the principles o
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Balb/c mice. The models described b
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urns-sepsis model, but not in the n
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ORF ID Gene name Gene function PA14
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functional activities it could be s
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Figure 1-6 Schematic showing the OR
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RhlI Autoinducer RhlR Rhl Regulon G
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1.5.2 Pseudomonas aeruginosa LES Th
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2 Material and Methods 2 MATERIAL A
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2.1.2 Strains used Pseudomonas aeru
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Plasmids Plasmid Description Refere
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In the case of capture vector const
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Primer name Sequence Capture vector
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The relevant homology sequence was
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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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- Page 187 and 188: AQ: 40 AQ: 41 DISCUSSION tapraid5/z
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- Page 193: 8 Bibliography AL-ALOUL, M., CRAWLE
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