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  • recA mediated spontaneous deletions of the icaADBC operon of clinical Staphylococcus epidermidis isolates: a new mechanism of phenotypic variations.

recA mediated spontaneous deletions of the icaADBC operon of clinical Staphylococcus epidermidis isolates: a new mechanism of phenotypic variations.

Antonie van Leeuwenhoek (2008-05-06)
Titik Nuryastuti, Henny C van der Mei, Henk J Busscher, Roel Kuijer, Abu T Aman, Bastiaan P Krom
RESUMO

Phenotypic variation of Staphylococcus epidermidis involving the slime related ica operon results in heterogeneity in surface characteristics of individual bacteria in axenic cultures. Five clinical S. epidermidis isolates demonstrated phenotypic variation, i.e. both black and red colonies on Congo Red agar. Black colonies displayed bi-modal electrophoretic mobility distributions at pH 2, but such phenotypic variation was absent in red colonies of the same strain as well as in control strains without phenotypic variation. All red colonies had lost ica and the ability to form biofilms, in contrast to black colonies of the same strain. Real time PCR targeting icaA indicated a reduction in gene copy number within cultures exhibiting phenotypic variation, which correlated with phenotypic variations in biofilm formation and electrophoretic mobility distribution of cells within a culture. Loss of ica was irreversible and independent of the mobile element IS256. Instead, in high frequency switching strains, spontaneous mutations in lexA were found which resulted in deregulation of recA expression, as shown by real time PCR. RecA is involved in genetic deletions and rearrangements and we postulate a model representing a new mechanism of phenotypic variation in clinical isolates of S. epidermidis. This is the first report of S. epidermidis strains irreversibly switching from biofilm-positive to biofilm-negative phenotype by spontaneous deletion of icaADBC.

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Sigma-Aldrich
Lysostaphin from Staphylococcus staphylolyticus, lyophilized powder, Protein 50-70 % by biuret, ≥500 units/mg protein
Sigma-Aldrich
Congo Red, Dye content ≥35 %
Sigma-Aldrich
Congo Red, certified by the Biological Stain Commission, BioXtra
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Lysostaphin from Staphylococcus simulans, recombinant, expressed in E. coli, lyophilized powder
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Lysostaphin from Staphylococcus staphylolyticus, BioUltra, ≥97% (SDS-PAGE), Protein 40-60 % by biuret, ≥2,000 units/mg protein
Supelco
Congo Red, analytical standard
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Lysostaphin from Staphylococcus staphylolyticus, aseptically filled