Location of Repository

Characterisation of pathogenicity islands in vivo and in vitro and the in vivo virulence of Pseudomonas aeruginosa

By Melissa Elvira Koreen Carter


Pseudomonas aeruginosa is an opportunistic pathogen and usually targets immunocompromised patients such as burn victims and patients with AIDS, cancer or cystic fibrosis (CF). Acquisition of this organism is associated with high mortality and can cause death within 24 hours. The main themes covered within this thesis are pathogenicity island characterisation in vitro and in vivo as well as investigation of in vivo virulence of P. aeruginosa. The rationale behind this focus is that 10-20% of the P. aeruginosa genome is variable between strains and large variable regions such as genomic (pathogenicity) islands are considered more likely to contribute to the differences in disease-causing ability between strains.\ud The first project covers the development of a novel generic yeast-based genomic island capture method, which enables a complete genomic island to be present within a cloning vector. It was used in the characterisation of genomic islands in both P. aeruginosa and Escherichia coli. A novel genomic island in E. coli was captured and characterised.\ud The second project investigates the contribution of two pathogenicity islands, PAPI- 1 and PAPI-2 to the in vivo virulence of P. aeruginosa PA14. Three pathogenicity island deletant isogenic mutants were tested for virulence in a murine acute respiratory model of infection developed for this project. The results showed that both pathogenicity islands contribute to virulence, but the presence of PAPI-2 is enough to maintain wild-type virulence.\ud The third project covers the exploration of the role of quorum-sensing in the virulence of P. aeruginosa LES; one of the transmissible epidemic strains and the most common strain recovered from CF patients across the UK. The project assessed whether over-expression of quorum-sensing products is a reliable indicator of increased virulence within a murine acute respiratory model of infection. The results showed in general, over-expressing mutants were more virulent than deficient mutants, but there was one exception, LESB58

Publisher: University of Leicester
Year: 2009
OAI identifier: oai:lra.le.ac.uk:2381/7578

Suggested articles



  1. (2005). A cystic fibrosis epidemic strain of Pseudomonas aeruginosa displays enhanced virulence and antimicrobial resistance.
  2. (2001). A functional genomic analysis of type 3 Streptococcus pneumoniae virulence. Mol Microbiol
  3. (2003). A general cloning system to selectively isolate any eukaryotic or prokaryotic genomic region in yeast.
  4. (2001). A genomic island in Pseudomonas aeruginosa carries the determinants of flagellin glycosylation.
  5. (2005). A guild of 45 CRISPR-associated (Cas) protein families and multiple CRISPR/Cas subtypes exist in prokaryotic genomes.
  6. A molecular sensor that allows a gut commensal to control its nutrient foundation in a competitive ecosystem.
  7. (1987). A mouse model of chronic pulmonary infection with Pseudomonas aeruginosa and Pseudomonas cepacia.
  8. (2003). A novel strategy for analysis of gene homologues and segmental genome duplications.
  9. (2006). A novel strategy for the identification of genomic islands by comparative analysis of the contents and contexts of tRNA sites in closely related bacteria.
  10. (2008). a rat model of chronic lung infection.
  11. (1979). A rat model of chronic respiratory infection with Pseudomonas aeruginosa.
  12. (1989). A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae.
  13. (2006). Acquisition and evolution of the exoU locus in Pseudomonas aeruginosa.
  14. (2000). Acquisition of expression of the Pseudomonas aeruginosa ExoU cytotoxin leads to increased bacterial virulence in a murine model of acute pneumonia and systemic spread.
  15. (2005). Activities of Pseudomonas aeruginosa effectors secreted by the Type III secretion system in vitro and during infection.
  16. (2004). An adenylate cyclasecontrolled signaling network regulates Pseudomonas aeruginosa virulence in a mouse model of acute pneumonia.
  17. (2005). An improved method for rapid generation of unmarked Pseudomonas aeruginosa deletion mutants.
  18. (2003). an orthologue of Mga, acts as a transcriptional repressor of the genes within the rlrA pathogenicity islet in Streptococcus pneumoniae.
  19. Analysis of the hypervariable region of the Salmonella enterica genome associated with tRNA(leuX).
  20. (2008). Analyzing real-time PCR data by the comparative C(T) method. Nat Protoc
  21. (2004). Antibacterial activity of a competence-stimulating peptide in experimental sepsis caused by Streptococcus pneumoniae.Antimicrob Agents Chemother
  22. (2004). Bacteriophage P4 Vis protein is needed for prophage excision.
  23. (2004). Biofilm formation by the small colony variant phenotype of Pseudomonas aeruginosa.
  24. (1997). BrandMiller JC,McNeil Y,McVeagh P. Sialic acid content of infant saliva: comparison of breast fedwith formula fed infants. ArchDis Child
  25. (2005). Cloning and analysis of specific Trypanosoma brucei TREU 927 telomeres by TAR cloning strategy,
  26. (1988). Cloning and expression of the pneumococcal neuraminidase gene in Escherichia coli. Gene
  27. (2000). Combination therapy for chronic Pseudomonas aeruginosa respiratory infection associated with biofilm formation.
  28. (1995). Common virulence factors for bacterial pathogenicity in plants and animals.
  29. (2007). Comparison of the composition of oral mucosal residual saliva with whole saliva. Oral Dis
  30. (2000). Complete genome sequence of Pseudomonas aeruginosa PA01, an opportunistic pathogen.
  31. (2003). Conservation of genome content and virulence determinants among clinical and environmental isolates of Pseudomonas aeruginosa.
  32. (1983). Construction of artificial chromosomes in yeast.
  33. (1991). Construction of versatile low-copynumber vectors for cloning, sequencing and gene expression in Escherichia coli.
  34. (1999). Contribution of quorum sensing to the virulence of Pseudomonas aeruginosa in burn wound infections.
  35. (1996). Contribution of specific Pseudomonas aeruginosa virulence factors to pathogenesis of pneumonia in a neonatal mouse model of infection.
  36. (2005). CRISPR elements in Yersinia pestis acquire new repeats by preferential uptake of bacteriophage DNA, and provide additional tools for evolutionary studies.
  37. (2007). CRISPR provides acquired resistance against viruses in prokaryotes.
  38. (2008). CRISPR--a widespread system that provides acquired resistance against phages in bacteria and archaea.
  39. (2004). Current protocols in molecular biology / edited by Frederick M.Ausubel ...[et al.],
  40. (1963). Decrease of bound sialic acid and inhibitor in chorioallantoic membranes infected with influenza virus.
  41. (2006). Deglycosylation of human glycoconjugates by the sequential activities of exoglycosidases expressed by Streptococcus pneumoniae.Mol Microbiol
  42. (1998). Designer deletion strains derived from Saccharomyces cerevisiae S288C: a useful set of strains and plasmids for PCR-mediated gene disruption and other applications.
  43. (2006). Differential expression of key pneumococcal virulence genes in vivo.Microbiology
  44. (2006). Direct detection of bacterial biofilms on the middle-ear mucosa of children with chronic otitis media. JAMA
  45. (1992). Direct transfer of plasmid DNA from yeast to E.coli by electroporation.
  46. (2007). Diversity of the abundant pKLC102/PAGI-2 family of genomic islands in Pseudomonas aeruginosa.
  47. (2004). Effect of antiviral treatment on the outcome of secondary bacterial pneumonia after influenza.
  48. (2007). Environmental regulation of Pseudomonas aeruginosa PAO1 Las and Rhl quorum-sensing systems.
  49. (1980). Epidemiologic studies of Streptococcus pneumoniae in infants: acquisition, carriage, and infection during the first 24 months of life.
  50. (2000). Evaluation of the virulence of a Streptococcus pneumoniae neuraminidase-deficient mutant in nasopharyngeal colonization and development of otitis media in the chinchilla model. Infect Immun
  51. Evidence for diversifying selection at the pyoverdine locus of Pseudomonas aeruginosa.
  52. (2006). Evolving stealth: genetic adaptation of Pseudomonas aeruginosa during cystic fibrosis infections.
  53. (2000). ExoT of cytotoxic Pseudomonas aeruginosa prevents uptake by corneal epithelial cells.
  54. (1997). ExoU expression by Pseudomonas aeruginosa correlates with acute cytotoxicity and epithelial injury.
  55. (2002). Gene islands integrated into tRNA(Gly) genes confer genome diversity on a Pseudomonas aeruginosa clone.
  56. (2006). Genetic adaptation by Pseudomonas aeruginosa to the airways of cystic fibrosis patients.
  57. (2001). Genetic transformation of yeast.
  58. (2002). Genetic variation at the O-antigen biosynthetic locus in Pseudomonas aeruginosa.
  59. (2004). Genome of bacteriophage P1.
  60. (2006). Genomic analysis reveals that Pseudomonas aeruginosa virulence is combinatorial.
  61. (2008). Growth of Streptococcus pneumoniae on human glycoconjugates is dependent upon the sequential activity of bacterial exoglycosidases.
  62. (2007). Growth phenotypes of Pseudomonas aeruginosa lasR mutants adapted to the airways of cystic fibrosis patients.
  63. (1985). Guidelines on the recognition of pain, distress and discomfort in experimental animals and an hypothesis for assessment.
  64. (2005). Heparin stimulates Staphylococcus aureus biofilm formation. Infect Immun
  65. (1996). Highly selective isolation of human DNAs from rodent-human hybrid cells as circular yeast artificial chromosomes by transformation-associated recombination cloning.
  66. (2002). Host cellular immune response to pneumococcal lung infection inmice.
  67. (2006). How phosphotransferase system-related protein phosphorylation regulates carbohydrate metabolism in bacteria. Microbiol Mol Biol Rev
  68. (2006). How to become a uropathogen: comparative genomic analysis of extraintestinal pathogenic Escherichia coli strains.
  69. (2004). Human and avian influenza viruses target different cell types in cultures of human airway epithelium. Proc Natl Acad Sci USA
  70. Hypersusceptibility of cystic fibrosis mice to chronic Pseudomonas aeruginosa oropharyngeal colonization and lung infection.
  71. (2004). Identification and characterization of transmissible Pseudomonas aeruginosa strains in cystic fibrosis patients in England and Wales.
  72. (2001). Identification of a genomic island present in the majority of pathogenic isolates of Pseudomonas aeruginosa.
  73. (2002). Identification of genes that are associated with DNA repeats in prokaryotes.
  74. (2002). Identification of virulence genes in a pathogenic strain of Pseudomonas aeruginosa by representational difference analysis.
  75. (2007). Impact of Pseudomonas aeruginosa quorum sensing on biofilm persistence in an in vivo intraperitoneal foreign-body infection model.
  76. (2004). Inhibition ofmicrobial sialidases—What happened beyond the influenza virus? Curr Med Chem- Anti-infective Agents
  77. (1976). Initial colonization of teeth in monkeys as related to diet. Infect Immun
  78. (2004). Integrated regulatory responses of fimB to N-acetylneuraminic (sialic) acid and GlcNAc
  79. (2006). Interstrain transfer of the large pathogenicity island (PAPI-1) of Pseudomonas aeruginosa.
  80. (2001). Introduction of DNA into yeast cells.
  81. (2001). Introduction of plasmid DNA into cells. Current protocols in molecular biology / edited by Frederick M.Ausubel ...[et al.],
  82. (2006). Keeping their options open: acute versus persistent infections.
  83. (1999). Killing of Caenorhabditis elegans by Pseudomonas aeruginosa used to model mammalian bacterial pathogenesis.
  84. (2002). Large-scale identification of serotype 4 Streptococcus pneumoniae virulence factors. Mol Microbiol
  85. (1998). Large-scale identification of virulence genes from Streptococcus pneumoniae. Infect Immun
  86. (2002). Lethal synergism between influenza virus and Streptococcus pneumoniae: characterization of a mouse model and the role of platelet-activating factor receptor.
  87. (2002). Linker-mediated recombinational subcloning of large DNA fragments using yeast.
  88. (2004). Method for introducing specific and unmarkedmutations into the chromosome of Streptococcus pneumoniae.Mol Biotechnol
  89. (2004). Microarray analysis of pneumococcal gene expression during invasive disease. Infect Immun
  90. (2007). MobilomeFINDER: web-based tools for in silico and experimental discovery of bacterial genomic islands. Nucleic acids research,
  91. (2004). Monitoring dynamic expression of nuclear genes in Chlamydomonas reinhardtii by using a synthetic luciferase reporter gene.
  92. (2000). Mouse models of chronic lung infection with Pseudomonas aeruginosa: models for the study of cystic fibrosis.
  93. (2002). Murine models of chronic Pseudomonas aeruginosa lung infection.
  94. (2006). Mutational analysis of RetS, an unusual sensor kinase-response regulator hybrid required for Pseudomonas aeruginosa virulence.
  95. (2003). NEBcutter: A program to cleave DNA with restriction enzymes.
  96. (2009). Newly introduced genomic prophage islands are critical determinants of in vivo competitiveness in the Liverpool Epidemic Strain of Pseudomonas aeruginosa.
  97. (2004). Nontypeable Haemophilus influenzae strain2019producesabiofilmcontainingN-acetylneuraminicacid thatmaymimic sialylated O-linked glycans. Infect Immun
  98. okay that I changed “Transonic 460, 35 kHz” to “sonicator water bath (a Transonic 460/H ultrasonic bath [Elma])”? 15. Au: (a) In the sentence “The sugars
  99. (2003). Optimum conditions for selective isolation of genes from complex genomes by transformation-associated recombination cloning.
  100. (2000). Pathogenicity islands and the evolution of microbes.
  101. (1998). PepA, a secreted protein of Pseudomonas aeruginosa, is necessary for cytotoxicity and virulence.
  102. (2004). Phase variable desialylation of host proteins that bind to Streptococcus pneumoniae in vivo and protect the airway. Mol Microbiol
  103. (2000). Plants and animals share functionally common bacterial virulence factors.
  104. (2006). Pneumococcal neuraminidases A and B both have essential roles during infection of the respiratory tract and sepsis. Infect Immun
  105. (2000). Positive correlation between virulence of Pseudomonas aeruginosa mutants in mice and insects.
  106. (1996). Posttranscriptional regulation of EcoP1I and EcoP15I restriction activity.
  107. (1996). Predominant pathogens found in the European Prevalence of Infection in Intensive Care Study. European journal of clinical microbiology & infectious diseases : official publication of the
  108. (2001). Preparation of genomic DNA from bacteria. Current protocols in molecular biology / edited by Frederick M.Ausubel ...[et al.],
  109. (2006). Presence of the exoU gene of Pseudomonas aeruginosa is correlated with cytotoxicity in MDCK cells but not with colonization in BALB/c mice.
  110. (2000). Primer3 on the WWW for general users and for biologist programmers.
  111. (2003). Prophage insertion sites.
  112. (2000). Pseudomonas aeruginosa cell-to-cell signaling is required for virulence in a model of acute pulmonary infection.
  113. (2007). Pseudomonas aeruginosa chronic colonization in cystic fibrosis patients. Current opinion in pediatrics,
  114. Pseudomonas aeruginosa cupA-encoded fimbriae expression is regulated by a GGDEF and EAL domain-dependent modulation of the intracellular level of cyclic diguanylate.
  115. (1999). Pseudomonas aeruginosa gene products PilT and PilU are required for cytotoxicity in vitro and virulence in a mouse model of acute pneumonia.
  116. (2003). Pseudomonas aeruginosa pneumonia. Current opinion in infectious diseases,
  117. (2004). Pseudomonas aeruginosa pyocyanin is critical for lung infection in mice.
  118. (2002). Pseudomonas biofilm formation and antibiotic resistance are linked to phenotypic variation.
  119. (1977). Purification and properties of the P15 specific restriction endonuclease from Escherichia coli.
  120. (1992). Recombination between similar but not identical DNA sequences during yeast transformation occurs within short stretches of identity.
  121. (1997). Recombination-mediated PCR-directed plasmid construction in vivo in yeast.
  122. (2004). Relative contributions of Pseudomonas aeruginosa ExoU, ExoS, and ExoT to virulence in the lung.
  123. (2003). Restriction endonucleases: classification, properties, and applications.
  124. (2003). Role of neuraminidase in lethal synergism between influenza virus and Streptococcus pneumoniae.
  125. (2005). Role of Pseudomonas aeruginosa quorum-sensing systems in a mouse model of chronic respiratory infection.
  126. (2006). Saccharomyces cerevisiae-based molecular tool kit for manipulation of genes from gram-negative bacteria.
  127. (2003). Secretion of the toxin ExoU is a marker for highly virulent Pseudomonas aeruginosa isolates obtained from patients with hospital-acquired pneumonia.
  128. (2006). Selective isolation of mammalian genes by TAR cloning.
  129. (2004). Sequence analysis of the mobile genome island pKLC102 of Pseudomonas aeruginosa C.
  130. (2007). serogroup-related differences in observed durations of nasopharyngeal carriage of penicillin-resistant pneumococci.
  131. (2008). Sialic acid metabolism and regulation by Haemophilus influenzae: potential novel antimicrobial therapies. Curr Infect Dis Rep
  132. (2007). Spec Care Dentist
  133. (2002). Spread of an epidemic Pseudomonas aeruginosa strain from a patient with cystic fibrosis (CF) to non-CF relatives.
  134. (1991). Streptococcus pneumoniae produces at least two distinct enzymes with neuraminidase activity: cloning and expression of a second neuraminidase gene in Escherichia coli. Infect Immun
  135. (2008). Structural basis of Escherichia coli single-stranded DNA-binding protein stimulation of exonuclease I.
  136. (2006). Studies on pneumolysin and on pneumococcal gene expression under aerobiosis.
  137. (2007). Sucrose metabolism contributes to in vivo fitness of Streptococcus pneumoniae.Mol Microbiol
  138. (2005). SudhakaraBabuY,McCullers JA. The novel parainfluenza virus hemagglutinin-neuraminidase inhibitor BCX 2798 prevents lethal synergism between a paramyxovirus and Streptococcus pneumoniae. Antimicrob Agents Chemother
  139. (2001). Superinfection with a transmissible strain of Pseudomonas aeruginosa in adults with cystic fibrosis chronically colonised by P aeruginosa.
  140. (2006). Switch from planktonic to sessile life: a major event in pneumococcal pathogenesis. MolMicrobiol
  141. (2001). Synonymous codon usage in bacteria.
  142. (2004). tapraid5/z9d-jid/z9d-jid/z9d01009/z9d4168d09a sangreyj S5 3/20/09 Art: 42180
  143. (2008). The Ashwell receptor mitigates the lethal coagulopathy of sepsis. Nat Med
  144. (2006). The association of respiratory viruses, temperature and other climatic parameters with the incidence of invasive pneumococcal disease in Sydney, Australia. Clin Infect Dis
  145. (2004). The broad host range pathogen Pseudomonas aeruginosa strain PA14 carries two pathogenicity islands harboring plant and animal virulence genes.
  146. (2006). The current burden of pneumococcal disease in England and Wales.
  147. (2006). The prevalence of penicillin-non-susceptible Streptococcus pneumoniae among children aged 5 years correlates with the biannual epidemic activity of respiratory syncytial virus. Clin Microbiol Infect
  148. (2002). The pyocins of Pseudomonas aeruginosa.
  149. (2008). The role of Streptococcus pneumoniae virulence factors in host respiratory colonization and disease. Nat Rev Microbiol
  150. (2007). Thewar against influenza: discovery and development of sialidase inhibitors. Nat Rev Drug Discov
  151. (2007). Towards pathogenomics: a web-based resource for pathogenicity islands. Nucleic acids research, 35(Database issue),
  152. (2002). Transformation of yeast by lithium acetate/single-stranded carrier DNA/polyethylene glycol method.
  153. (1987). Transformation of yeast spheroplasts without cell fusion.
  154. (2002). Tuberculosis drug targets.
  155. (2004). Tuomanen EI. Tissue-specific contributions of pneumococcal virulence factors to pathogenesis. J Infect Dis
  156. (2004). Twenty year surveillance of invasive pneumococcal disease in Nottingham: serogroups responsible and implications for immunisation. Arch Dis Child
  157. (2002). Type III protein secretion is associated with poor clinical outcomes in patients with ventilator-associated pneumonia caused by Pseudomonas aeruginosa.
  158. (2001). update, how to gapture.
  159. (2007). update, protocol for yeast recombinational cloning. Available: http://mml.sjtu.edu.cn/MobilomeFINDER/YCV_data/Protocol_for_Yeast_Recombi national_cloning.pdf.
  160. (2006). Use of suppression subtractive hybridization to examine the accessory genome of the Liverpool cystic fibrosis epidemic strain of Pseudomonas aeruginosa.
  161. (2008). UvrD and UvrD252 counteract RecQ, RecJ and RecFOR in the rep mutant.
  162. (2002). Ventilator-associated pneumonia. American journal of respiratory and critical care medicine,
  163. (2007). Widespread pyocyanin over-production among isolates of a cystic fibrosis epidemic strain.
  164. (1998). Yeast physiology and biotechnology.
  165. (1981). Yeast transformation: a model system for the study of recombination.

To submit an update or takedown request for this paper, please submit an Update/Correction/Removal Request.