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Genetic engineering of untransformable coagulase-negative staphylococcal pathogens

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Genetic engineering of untransformable coagulase-negative staphylococcal pathogens. / Winstel, Volker; Kühner, Petra; Rohde, Holger; Peschel, Andreas.

In: NAT PROTOC, Vol. 11, No. 5, 05.2016, p. 949-59.

Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review

Harvard

Winstel, V, Kühner, P, Rohde, H & Peschel, A 2016, 'Genetic engineering of untransformable coagulase-negative staphylococcal pathogens', NAT PROTOC, vol. 11, no. 5, pp. 949-59. https://doi.org/10.1038/nprot.2016.058

APA

Winstel, V., Kühner, P., Rohde, H., & Peschel, A. (2016). Genetic engineering of untransformable coagulase-negative staphylococcal pathogens. NAT PROTOC, 11(5), 949-59. https://doi.org/10.1038/nprot.2016.058

Vancouver

Winstel V, Kühner P, Rohde H, Peschel A. Genetic engineering of untransformable coagulase-negative staphylococcal pathogens. NAT PROTOC. 2016 May;11(5):949-59. https://doi.org/10.1038/nprot.2016.058

Bibtex

@article{f5a4c97b822641e7a65262bf5f787520,
title = "Genetic engineering of untransformable coagulase-negative staphylococcal pathogens",
abstract = "Coagulase-negative staphylococci (CoNS) are recognized as significant opportunistic pathogens. However, current knowledge of virulence mechanisms is very limited because a significant proportion of CoNS are refractory to available techniques for DNA transformation. We describe an efficient protocol for plasmid transfer using bacteriophage Φ187, which can transduce plasmid DNA to a wide range of CoNS from a unique, engineered Staphylococcus aureus strain. The use of a restriction-deficient, modification-proficient S. aureus PS187 mutant, which has a CoNS-type bacteriophage surface receptor, allows plasmid transfer to CoNS even when they are refractory to electroporation. Once the Φ187 titer reaches 10(9) plaque-forming units per milliliter, plasmid transfer can be accomplished within 1-2 d. Thus, our protocol is a major technical advance offering attractive opportunities for research on CoNS-mediated infections.",
author = "Volker Winstel and Petra K{\"u}hner and Holger Rohde and Andreas Peschel",
year = "2016",
month = may,
doi = "10.1038/nprot.2016.058",
language = "English",
volume = "11",
pages = "949--59",
journal = "NAT PROTOC",
issn = "1754-2189",
publisher = "NATURE PUBLISHING GROUP",
number = "5",

}

RIS

TY - JOUR

T1 - Genetic engineering of untransformable coagulase-negative staphylococcal pathogens

AU - Winstel, Volker

AU - Kühner, Petra

AU - Rohde, Holger

AU - Peschel, Andreas

PY - 2016/5

Y1 - 2016/5

N2 - Coagulase-negative staphylococci (CoNS) are recognized as significant opportunistic pathogens. However, current knowledge of virulence mechanisms is very limited because a significant proportion of CoNS are refractory to available techniques for DNA transformation. We describe an efficient protocol for plasmid transfer using bacteriophage Φ187, which can transduce plasmid DNA to a wide range of CoNS from a unique, engineered Staphylococcus aureus strain. The use of a restriction-deficient, modification-proficient S. aureus PS187 mutant, which has a CoNS-type bacteriophage surface receptor, allows plasmid transfer to CoNS even when they are refractory to electroporation. Once the Φ187 titer reaches 10(9) plaque-forming units per milliliter, plasmid transfer can be accomplished within 1-2 d. Thus, our protocol is a major technical advance offering attractive opportunities for research on CoNS-mediated infections.

AB - Coagulase-negative staphylococci (CoNS) are recognized as significant opportunistic pathogens. However, current knowledge of virulence mechanisms is very limited because a significant proportion of CoNS are refractory to available techniques for DNA transformation. We describe an efficient protocol for plasmid transfer using bacteriophage Φ187, which can transduce plasmid DNA to a wide range of CoNS from a unique, engineered Staphylococcus aureus strain. The use of a restriction-deficient, modification-proficient S. aureus PS187 mutant, which has a CoNS-type bacteriophage surface receptor, allows plasmid transfer to CoNS even when they are refractory to electroporation. Once the Φ187 titer reaches 10(9) plaque-forming units per milliliter, plasmid transfer can be accomplished within 1-2 d. Thus, our protocol is a major technical advance offering attractive opportunities for research on CoNS-mediated infections.

U2 - 10.1038/nprot.2016.058

DO - 10.1038/nprot.2016.058

M3 - SCORING: Journal article

C2 - 27101516

VL - 11

SP - 949

EP - 959

JO - NAT PROTOC

JF - NAT PROTOC

SN - 1754-2189

IS - 5

ER -