Loss of the Penicillinase Plasmid After Treatment of Staphylococcus aureus with Sodium Dodecyl Sulfate

Abstract

Two strains of Staphylococcus aureus (PC1 and 196E), when grown in medium containing 0.002% sodium dodecyl sulfate (SDS), showed from 96.1 to 100% loss of the ability to produce penicillinase. Resistance to cadmium and zinc was lost concomitantly with the ability to produce penicillinase. A comparison of the rate of curing by SDS with the curing effects exerted by elevated temperature and ethidium bromide suggested that SDS is a more effective plasmid curing agent for susceptible strains of S. aureus than methods in use at present.

Nature of the Genetic Determinant Controlling Exfoliative Toxin Production in Staphylococcus aureus

“…The early and rapid rate of appearance of Toxcells during growth of the Tox+ culture in either EB or SDS, or at 44 C, and the percentage of Tox-variants appearing in the population with time indicate that the selection and growth of a small number of spontaneously produced Toxcells could not account for the high frequencies of negative variants which appeared during the early intervals of growth during treatment. This is supported further by the observation that a cured substrain has no selective growth advantage over a similarly treated but uncured derivative when both substrains are grown again in either EB or SDS, or at 44 C. Similar data have been presented to provide evidence for the extrachromosomal nature of other staphylococcal genes (9,14,16,17,19).…”

“…A second chemical agent successfully used to eliminate staphylococcal plasmids was sodium dodecyl sulfate. Growth of two strains of S. aureus in 0.002% sodium dodecyl sulfate resulted in high rates of loss of ability to produce penicillinase (19). Loss of some, but not all, staphylococcal plasmids can be enhanced by growth at elevated temperatures (9,16).…”

“…The resulting colonies were examined for loss of ET, cadmium resistance, and penicillin resistance. The method of Sonstein and Baldwin (19) was used for plasmid elimination with SDS. An initial inoculum of 105 cells per ml was incubated with shaking at 37 C for 18 h in P broth containing 0.003% SDS.…”

“…The resulting phage concentration was approximately 109 to 1010 PFU/ml. Transduction was performed by the method of Sonstein and Baldwin (19) with the exception of P broth being used in place of Trypticase soy broth, and P agar without CaCl, in place of yeast extract-Trypticase soy agar. The selective concentration of CdNO, was 90 Mg/ml for recipient strain 04081 and 60 Mg/ml for recipient strain UT 0017, and the selective concentration of penicillin was 1.6 U/ml for both recipients.…”

Strategies for Developing CRISPR‐Based Gene Editing Methods in Bacteria

“…Some earlier studies indicated that supplementing low concentrations of sodium dodecyl sulfate (SDS) or ethidium bromide (EB) would facilitate the natural loss of plasmid in S. aureus. [150,151] However, the methods lack further exploration in other bacterial species and rise the possible secondary loss of other intrinsic plasmids. To selectively and efficiently cure the artificial plasmids in bacteria, temperature-sensitive (TS) replicons are widely used in many genera (Table S1, Supporting Information), whose replication is generally allowed at a relatively low temperature (typically around 30 °C).…”