Transformation analysis of three linkage groups in Staphylococcus aureus

Abstract

While studying a set of multiply marked mutants of Staphylococcus aureus strain 8325 by transformation, several instances of apparent genetic linkage were encountered. After showing that these linked transformations were readily inactivated by shearing of the deoxyribonucleic acid (DNA) but were resistant to dilution of the DNA, and showing that mixtures of DNA failed to form double transformants, it was concluded that the linkages were legitimate rather than the result of congression. Three linkage groups were defined: thy-101-lys-115-trp-103-thr-106, pyr-141-hisGb15-nov-pur-102, and pur-110-ilv-129. The positions of the previously studied trp and his operons corresponded to the trp-103 and hisGbl5 loci. The ilv-129 position adjacent to pur-l10 probably corresponds to the ilv-leu gene cluster. The distance over which linkage was detected was greater by transformation than by generalized transduction. 201 on August 11, 2020 by guest http://jb.asm.org/ Downloaded from for 30 min on a Burrell wrist-action shaker (Burrell distilled (under N2) phenol saturated with 0.01 Corp., Pittsburgh, Pa.). The lysis mixtures were then M Tris(hydroxymethylfaminomethane-hydrochloride extracted three times with equal volumes of freshly buffer, pH 8.

Polyamine‐independent growth and biofilm formation, and functional spermidine/spermine N‐acetyltransferases in Staphylococcus aureus and Enterococcus faecalis

“…For planktonic growth, S. aureus SH1000 (Horsburgh et al, 2002;O'Neill, 2010), a methicillin sensitive S. aureus was grown in Tryptic Soy Broth (TSB) or polyamine-free chemically defined medium (PN medium) (Pattee and Neveln, 1975) at 37°C, buffered to pH 7.0 from single colonies. The S. aureus SH1000 strain is derived from NCTC 8325 that had been cured of its three prophages to yield strain 8325-4, and thereafter the 8325-4 strain rsbU gene was repaired to yield strain SH1000 (Horsburgh et al, 2002).…”

Arginine catabolic mobile element encoded speG abrogates the unique hypersensitivity of Staphylococcus aureus to exogenous polyamines

“…Strains used in this work are listed in Table S1. Bacteria were generally cultivated in BHI Broth/Agar (Difco, Detroit, MI) or chemically defined PN media (Pattee and Neveln, 1975) in shaking cultures (250 r.p.m.) at 37°C.…”

THE STAPHYLOCOCCAL ENTEROTOXINS — A GENETIC OVERVIEW1

“…Recently, Pattee and Glatz (1980) attempted to transform the e n t E phenotype in strain S6 and S. aureus C234. However since these determinants were apparently located outside the three linkage groups of known staphylococcal genes (see Pattee and Neveln 1975 for details of these mapping procedures), they were unable to convert a set of multiply-marked auxotrophic derivatives of NCTC 8325 to SEB' and the location of the genes governing SEB could not be determined. Even if they were successful, these experiments would be difficult to evaluate since our laboratory has shown that two determinants (pSN2 and the presumed SEB structural gene) are required for enterotoxigenesis.…”