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J Bacteriol. 2017 May 30. pii: JB.00271-17. doi: 10.1128/JB.00271-17. [Epub ahead of print]

Bypassing the restriction system to improve transformation of Staphylococcus epidermidis.

Author information

1
Department of Microbiology and Immunology, Geisel School of Medicine at Dartmouth, Hanover, NH 03755.
2
Genomic Research Laboratory, Service of Infectious Diseases, University Hospital of Geneva, 1205 Geneva 4, Switzerland.

Abstract

Staphylococcus epidermidis is the leading cause of infections on indwelling medical devices worldwide. Intrinsic antibiotic resistance and vigorous biofilm production have rendered these infections difficult to treat and, in some cases, require the removal of the offending medical prostheses. With the exception of two widely-passaged isolates RP62A and 1457, the pathogenesis of infections caused by clinical S. epidermidis strains is poorly understood due to the strong genetic barrier that precludes efficient transformation of foreign DNA into clinical isolates. The difficulty in transforming clinical S. epidermidis isolates is primarily due to the type I and IV restriction modification systems which act as genetic barriers. Here, we showed that efficient plasmid transformation of clinical S. epidermidis isolates from clonal complexes 2, 10 and 89 could be realized by employing a plasmid artificial modification (PAM) in E. coli DC10B containing a Δdcm mutation. This transformative technique should facilitate our ability to genetically modify clinical isolates of S. epidermidis and hence improve our understanding of its pathogenesis in human infections.ImportanceStaphylococcus epidermidis is a source of considerable morbidity worldwide. The underlying mechanisms contributing to the commensal and pathogenic lifestyles of S. epidermidis are poorly understood. Genetic manipulations of clinically relevant strains of S. epidermidis are largely prohibited due to the presence of a strong restriction barrier. With the introductions of the tools presented here, genetic manipulation has now become possible with clinically relevant S. epidermidis isolates, thus improving our understanding of S. epidermidis as a pathogen.

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