Title page for ETD etd-12072000-151028


Type of Document Master's Thesis
Author Burnley, Leigh-Emma
Author's Email Address leighemmaburnley@aol.com
URN etd-12072000-151028
Title Heavy Metal Resistance in the Genus Gluconobacter
Degree Master of Science
Department Biology
Advisory Committee
Advisor Name Title
Claus, G. William Committee Chair
Popham, David L. Committee Member
Yousten, Allan A. Committee Member
Keywords
  • Gluconobacter
  • zinc
  • cadmium
  • plasmids
  • heavy metal resistance
  • Ralstonia
  • cobolt
  • czc
Date of Defense 2000-11-10
Availability unrestricted
Abstract
The genus Gluconobacter is industrially important due to the ability to accomplish unusual and almost complete oxidation reactions (bioconversions) and to contaminate high sugar content products. Following preliminary evidence that some strains of Gluconobacter were resistant to cadmium, and realizing that cadmium resistance among gram-negative organisms is often encoded by an operon which also encodes cobalt and zinc resistance via an efflux mechanism, 10 strains of Gluconobacter were tested for heavy-metal resistance. Three of the 10 representative strains appeared to be resistant to cadmium chloride, and two were also resistant to cobalt- and zinc chloride. These strains, as well as two cadmium-sensitive strains were analyzed using PCR and sequencing to establish gene homology with Ralstonia eutropha, the most frequently studied Gram-negative bacterium exhibiting cadmium resistance. Amplification of two genes from the czc operon, known to encode cadmium, cobalt and zinc resistance in Ralstonia, was attempted in the three resistant and two sensitive strains of Gluconobacter. The gene, czcA, thought to encode the main pump protein of the efflux mechanism, was found in all Gluconobacter strains tested. However, amplification of a regulatory gene czcD, thought to sense the extracellular metal ion concentration, was not possible in the Gluconobacter strains tested. The PCR products were sequenced and analyzed for homology to the czc operon in Ralstonia. From the data gathered, it appears as though some strains of Gluconobacter contain at least a portion of the czc operon , encoding cadmium, cobalt and zinc resistance in Ralstonia eutropha.
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