Electrotransformation of Thiobacillus ferrooxidans with plasmids containing a mer determinant

T. Kusano, K. Sugawara, C. Inoue, T. Takeshima, M. Numata, T. Shiratori

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49 Citations (Scopus)

Abstract

The mer operon from a strain of Thiobacillus ferrooxidans (C. Inoue, K. Sugawara, and T. Kusano, Mol. Microbiol. 5:2707-2718, 1991) consists of the regulatory gene merR and an operator-promoter region followed by merC and merA structural genes and differs from other known gram-negative mer operons. We have constructed four potential shuttle plasmids composed of a T. ferrooxidans-borne cryptic plasmid, a pUC18 plasmid, and the above-mentioned mer determinant as a selectable marker. Mercury ion-sensitive T. ferrooxidans strains were electroporated with constructed plasmids, and one strain, Y4-3 (of 30 independent strains tested), was found to have a transformation efficiency of 120 to 200 mercury-resistant colonies per μg of plasmid DNA. This recipient strain was confirmed to be T. ferrooxidans by physiological, morphological, and chemotaxonomical data. The transformants carried a plasmid with no physical rearrangements through 25 passages under no selective pressure. Cell extracts showed mercury ion-dependent NADPH oxidation activity.

Original languageEnglish
Pages (from-to)6617-6623
Number of pages7
JournalJournal of bacteriology
Volume174
Issue number20
DOIs
Publication statusPublished - 1992

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

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