Thesis (M.A.)--Central Connecticut State University, 2002. / Thesis advisor: Michael A. Davis. " ... in partial fulfillment of the requirements for the degree of Master of Arts in Biology." Includes bibliographical references. Also available via the World Wide Web.

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Listeria monocytogenes is a Gram-positive bacterium that is found ubiquitously throughout nature and is the etiologic agent of listeriosis. The majority of human listeriosis is foodborne, resulting from the consumption of unpasteurized and ready-to-eat foods that are contaminated during food processing. During the 2008 nationwide outbreak, the Gilmour laboratory performed the first real-time application of high-throughput whole-genome sequencing (WGS) of outbreak strains. Within this genomic data, the 77 kb plasmid, pLM5578, was newly identified in a clinical isolate, and additional Listeria plasmids (the 80 kb pLM5026 and the 60 kb pLM0813) were subsequently identified after WGS was completed on an expanded panel of outbreak isolates. Little was known regarding how plasmids contribute to persistence and virulence of L. monocytogenes, and to investigate these potential relationships, a panel of 147 L. monocytogenes food, environmental, and clinical isolates from Canadian public health events from the last three decades was selected for further study of the plasmids they might contain. Strain carriage of plasmids was determined using conventional PCR targeting known plasmid gene targets. Bioinformatic analyses were then used to predict the functions of individual genes encoded by each sequenced plasmid. These analyses were then used to direct experiments investigating the functions and associated phenotypes conferred by plasmid carriage. Phenotypic analyses included antimicrobial susceptibility testing, heavy metal resistance, and biofilm formation assays. Finally, WGS analyses was performed on isolates with plasmid screening patterns that indicated carriage of potential novel plasmids. Screening revealed that 75 of 147 isolates were positive for the presence of a plasmid, for which WGS analysis identified 24 unique newly identified L. monocytogenes plasmids. Phenotypically, 15 of these plasmids were found to contribute to a decreased susceptibility to the heavy metal cadmium, whereas 4 conferred resistance to the sanitizer benzalkonium chloride. Plasmid carriage was also found to affect biofilm formation. Nine plasmids correlated with stronger biofilm formation phenotypes; whereas 5 plasmids were correlated with weaker biofilm formation phenotypes. No known virulence factors or antibiotic resistance determinants were present in the DNA sequences of these 24 newly identified plasmids. Numerous coding sequences predicted to assist with survival under environmental stress were identified, and it is hypothesized that these plasmids likely contributed to persistence of L. monocytogenes within food processing environments.

Bibliography: leaves 113-130 / ix, 130 leaves, [28] leaves of plates : ill. (some col.) ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Thesis (Ph.D.)--University of Adelaide, Dept. of Plant Pathology, 1988

The initial aim of this project was to characterise the integrative recombination mechanism of bacteriophage ÖAR29 , to provide a better understanding for development of the shuttle plasmid pBA as a site-specific Bacteroides integration vector. RT-PCR showed that the previously identified ÖAR29 recombination genes, integrase (Int) and excisionase (Xis), were transcribed from pBA in E. coli SCS110, B. thetaiotaomicron AR29 and B. uniformis AR20. In silico derived amino acid sequences from both genes showed only very low levels of similarity to other known Int and Xis in GenBank. To improve understanding of the phage recombination system, the ÖAR29 genome was sequenced. This revealed a 35,558 bp double-stranded DNA genome with GC content of 39.11%. Bioinformatic analysis identified 53 open reading frames (>30 codons) and gene promoters and terminators that allowed the genome arrangement to be compared with other phages. Comparison of deduced gene products with proteins from other phages identified 6 reading frames, allowed tentative identification of 7 others, but left 40 ORFs unidentified. Those with strong homology to known genes were: large terminase subunit (44.66 kDa), dnaC (27.94 kDa), helix-turn-helix (HTH) transcription regulator (14.69 kDa), cI repressor (26.48 kDa), amidase (18.42kDa) and a novel integrase (54.22 kDa). The integrase gene is located 162 base-pairs downstream of the phage attachment (attP) core site, rather than the previously suggested location upstream of the integration site. The ÖAR29 attP was shown to include a 16-bp att core region, 117 bp upstream of the previously suggested location. Integration of ÖAR29 was found to occur at the 3’end of an arg-tRNA gene on the AR29 genome (attB). Imperfect direct repeats with a consensus sequence (ANGTTGTGCAA) were found surrounding the attP core. A review of pBA sequence showed that only the 5’ end (435 bp) of the newly identified Int gene was cloned in pBA. Despite this, PCR analysis revealed integration of pBA into the AR29 genome. Serial subculturing of pBA transformed AR29 was able to cure AR29 of the ÖAR29 prophage, providing an improved host for integrative plasmids, and for detailed studies of AR29 physiology and ÖAR29 life cycles.

Thesis (Ph.D.)--Murdoch University, 2005. / Thesis submitted to the Division of Science and Engineering. Bibliography: leaves 211-230.

Thesis (Ph. D.)--University of Hong Kong, 2009. / Includes bibliographical references (p. 143-183). Also available in print.