Journal of Antimicrobial Chemotherapy (2002) 50, 313-321
© 2002 The British Society for Antimicrobial Chemotherapy
Genetic characterization of the fusidic acid and cadmium resistance determinants of Staphylococcus aureus plasmid pUB101
1 School of Biomedical Sciences and Gram-positive Bacteria Typing and Research Unit, Curtin University of Technology, Perth 6845, Western Australia, Australia; 2 Microbiology Department, College of Osteopathic Medicine, Midwestern University, Downers Grove, IL 60515, USA
Received 7 March 2002; returned 7 May 2002; revised 14 June 2002; accepted 20 June 2002
We report the cloning of the fusidic acid and cadmium resistance determinants from Staphylococcus aureus plasmid pUB101. The pUB101 fusidic acid resistance determinant was located on a 2.9 kb HindIII fragment. Sequencing of this fragment revealed three putative open reading frames (ORFs) of 213 (far1), 152 (orf152) and 170 amino acids (orf170), which are flanked by the right-hand end of insertion sequence IS431/257 (IS431/257RH) and a partial ORF. Far1 and Orf152 demonstrated homology with a chromosomally encoded fibronectin-binding protein of Listeria monocytogenes and the putative protein YosT, found on the SPßc2 prophage of Bacillus subtilis, respectively. Transformation of S. aureus with a construct containing a 949 bp far1-specific amplicon led to the isolation of a fusidic acid-resistant transformant, thereby identifying the pUB101 fusidic acid resistance structural gene. Between orf152 and far1 we identified a unique 113 bp symmetrical element and other repeat elements that may be involved with the control of orf152 and/or far1 expression. Hybridization of Southern blots revealed that far1 was not located on the chromosome or plasmid content of a limited number of Australian, UK and Hong Kong fusidic acid-resistant isolates. The pUB101 cadmium resistance determinant was located on a 3.6 kb HindIII fragment that carried a cadDX operon, remnants of two putative plasmid replication protein genes and IS431/257RH. Sequence analysis also demonstrated the presence of a single-stranded origin of replication, normally found on rolling circle replicating plasmids, within the putative promoter region of the cadDX operon.
* Correspondence address. Department of Biology MSC 3AF, New Mexico State University, PO Box 30001, Las Cruces, NM 88003-8001, USA. Tel: +1-505-646-3611; Fax: +1-505-646-5660; E-mail: jgustafs{at}nmsu.edu
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