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PMID: 1400213 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Electrotransformation of Thiobacillus ferrooxidans with plasmids containing a mer determinant.

Journal of bacteriology ·Vol. 174 ·No. 20 ·1992-10-00 ·Pages 6617-23

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

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 microgram 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.

Related Genes
mer
MeSH Terms
Blotting, Southern Cloning, Molecular Drug Resistance, Microbial/genetics Escherichia coli/drug effects,genetics Genes, Bacterial/genetics Genetic Markers/genetics Genetic Vectors/genetics Mercury/pharmacology Plasmids/genetics Restriction Mapping Thiobacillus/drug effects,genetics Transformation, Bacterial/genetics
Chemicals
Genetic Markers Mercury
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Kusano T
Laboratory of Plant Genetic Engineering, Akita Prefectural College of Agriculture, Japan.
Sugawara K
Inoue C
Takeshima T
Numata M
Shiratori T
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.174.20.6617-6623.1992
Published
1992-10-00
Pages
6617-23
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC207636
Subset
IM
External Links
PubMed source DOI: 10.1128/jb.174.20.6617-6623.1992 PMC Full Text View journal details
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