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Identification and characterization of organotin inducible bacterial genes

Organotin compounds are used for a variety of applications including antifouling paints for ship hulls, wood preservatives, and UV stabilizers in plastics. When they are released into aquatic environments, they easily accumulate in various marine organisms. Organotin compounds are considered endocrine disrupting chemicals since numerous marine organisms exhibit sexual abnormalities when exposed to dibutyltin. Singe certain bacteria show resistance to these compounds, we screened for genes that are induced by organotins. 3000 Escherichia coli clones were genetically engineered to contain the luxAB reporter genes from Vibrio harveyi in single, yet different, chromosomal locations. We screened these 3000 clones in the presence and absence of organotins, and identified a uniquely responsive clone designated TBT3. Luminescence was measured in liquid media and in solid media. Upon addition of tributyltin and dibutyltin, luminescence greatly increased. The transcriptional activity was measured by RNA dot blotting, and two potential genes ygaP and b2667, were identified as induced by organotins. Also, TBT3 transcriptional activity was measured and compared with the transcriptional activity of MG1655, the parent strain of TBT3. The measured transcriptional activity of the two strains was different, although the conditions in which the measurements were taken were identical, which gives place to interesting arguments what determines this difference.

Identiferoai:union.ndltd.org:LACETR/oai:collectionscanada.gc.ca:QMM.80893
Date January 2003
CreatorsVitos, Gabriella
ContributorsDuBow, M. S. (advisor), Marczynski, G. T. (advisor)
PublisherMcGill University
Source SetsLibrary and Archives Canada ETDs Repository / Centre d'archives des thèses électroniques de Bibliothèque et Archives Canada
LanguageEnglish
Detected LanguageEnglish
TypeElectronic Thesis or Dissertation
Formatapplication/pdf
CoverageMaster of Science (Department of Microbiology and Immunology.)
RightsAll items in eScholarship@McGill are protected by copyright with all rights reserved unless otherwise indicated.
Relationalephsysno: 002032010, proquestno: AAIMQ98758, Theses scanned by UMI/ProQuest.

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