Affiliation:
1. Department of Microbiology and Immunology, Geisel School of Medicine at Dartmouth, Hanover, New Hampshire, USA
2. Genomic Research Laboratory, Service of Infectious Diseases, University Hospital of Geneva, Geneva, Switzerland
Abstract
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 prosthesis. 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 the 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 show that efficient plasmid transformation of clinical
S. epidermidis
isolates from clonal complexes 2, 10, and 89 can be realized by employing a plasmid artificial modification (PAM) in
Escherichia coli
DC10B containing a Δ
dcm
mutation. This transformative technique should facilitate our ability to genetically modify clinical isolates of
S. epidermidis
and hence improve our understanding of their pathogenesis in human infections.
IMPORTANCE
Staphylococcus 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 of clinically relevant
S. epidermidis
isolates has now become possible, thus improving our understanding of
S. epidermidis
as a pathogen.
Funder
NIAID
Swiss National Foundation
Publisher
American Society for Microbiology
Subject
Molecular Biology,Microbiology
Cited by
21 articles.
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