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. 2005 Aug;71(8):4935-7.
doi: 10.1128/AEM.71.8.4935-4937.2005.

Identification of genes involved in cytochrome c biogenesis in Shewanella oneidensis, using a modified mariner transposon

R Bouhenni  1 , A Gehrke, D Saffarini
Affiliations

Identification of genes involved in cytochrome c biogenesis in Shewanella oneidensis, using a modified mariner transposon

R Bouhenni et al. Appl Environ Microbiol. 2005 Aug.

Abstract

A modified mariner transposon, miniHimar RB1, was generated to mutagenize cells of the metal-reducing bacterium Shewanella oneidensis. The use of this transposon led to the isolation of stable mutants and allowed rapid identification of disrupted genes. Fifty-eight mutants, including BG104 and BG148 with transposon insertions in the cytochrome c maturation genes ccmC and ccmF1, respectively, were analyzed. Both mutants were deficient in anaerobic respiration and cytochrome c production.

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Figures

FIG. 1.
FIG. 1.
Map of pMiniHimar RB1. The locations of oriT-Plac upstream of the transposase gene, inverted repeats (IR), origin of replication (R6K), and kanamycin (Km) resistance gene are indicated.
FIG. 2.
FIG. 2.
Arrangement of the ccm genes on the S. oneidensis chromosome. Insertion sites of miniHimar RB1 (filled triangles) and the names of the resulting mutants are indicated.
FIG. 3.
FIG. 3.
Detection of c-type cytochromes in S. oneidensis strains. Proteins (100 μg) were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and then stained for heme-associated peroxidase activity. Lanes 1 through 3, cell extracts from aerobically grown wild type, BG104 (ccmF1 mutant), and BG148 (ccmC mutant); lanes 4 through 6, cell extracts from anaerobic wild type, BG104, and BG148.
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