Transfer of plasmid DNA to clinical coagulase-negative staphylococcal pathogens using a unique bacteriophage

TY - JOUR

T1 - Transfer of plasmid DNA to clinical coagulase-negative staphylococcal pathogens using a unique bacteriophage

AU - Winstel, Volker

AU - Kühner, Petra

AU - Krismer, Bernhard

AU - Peschel, Andreas

AU - Rohde, Holger

N1 - Copyright © 2015, American Society for Microbiology. All Rights Reserved.

PY - 2015/1/23

Y1 - 2015/1/23

N2 - Genetic manipulation of emerging bacterial pathogens such as coagulase-negative staphylococci (CoNS) is a major hurdle in clinical and basic microbiological research. Strong genetic barriers such as restriction modification systems or clustered regularly interspaced short palindromic repeats (CRISPR) usually interfere with available techniques for DNA transformation and therefore complicate or render manipulation of CoNS impossible. Thus, current knowledge on pathogenicity and virulence determinants of CoNS is very limited. Here, a rapid, efficient and highly reliable technique is presented to transfer plasmid DNA essential for genetic engineering to important CoNS pathogens from a unique Staphylococcus aureus strain via a specific S. aureus bacteriophage Φ187. Even strains refractory to electroporation can be transduced by this technique once donor and recipient strains share similar Φ187 receptor properties. As a proof of principle, this technique was used to delete the alternative transcription factor sigma B (SigB) via allelic replacement in nasal and clinical Staphylococcus epidermidis isolates at high efficiencies. The described approach will allow for the genetic manipulation of a wide range of CoNS pathogens and might inspire research activities to manipulate other important pathogens in a similar fashion.

AB - Genetic manipulation of emerging bacterial pathogens such as coagulase-negative staphylococci (CoNS) is a major hurdle in clinical and basic microbiological research. Strong genetic barriers such as restriction modification systems or clustered regularly interspaced short palindromic repeats (CRISPR) usually interfere with available techniques for DNA transformation and therefore complicate or render manipulation of CoNS impossible. Thus, current knowledge on pathogenicity and virulence determinants of CoNS is very limited. Here, a rapid, efficient and highly reliable technique is presented to transfer plasmid DNA essential for genetic engineering to important CoNS pathogens from a unique Staphylococcus aureus strain via a specific S. aureus bacteriophage Φ187. Even strains refractory to electroporation can be transduced by this technique once donor and recipient strains share similar Φ187 receptor properties. As a proof of principle, this technique was used to delete the alternative transcription factor sigma B (SigB) via allelic replacement in nasal and clinical Staphylococcus epidermidis isolates at high efficiencies. The described approach will allow for the genetic manipulation of a wide range of CoNS pathogens and might inspire research activities to manipulate other important pathogens in a similar fashion.

U2 - 10.1128/AEM.04190-14

DO - 10.1128/AEM.04190-14

M3 - SCORING: Journal article

C2 - 25616805

VL - 81

SP - 2481–2488.

JO - APPL ENVIRON MICROB

JF - APPL ENVIRON MICROB

SN - 0099-2240

ER -