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Cloning of the gene conferring resistance to mupirocin in Staphylococcus aureus
K G Dyke1, S P Curnock, M Golding
1Department of Biochemistry, University of Oxford, U.K.
Abstract:
A plasmid known to be associated with mupirocin resistance of Staphylococcus aureus has been isolated and a restriction enzyme map constructed. An EcoRI fragment of 4.05 kb from this plasmid has been cloned into an Escherichia coli-Staphylococcus aureus shuttle vector and shown to carry the gene for resistance to mupirocin. The DNA sequence of a small section of the gene has been determined and the derived amino acid sequence compared with a data bank. The amino acid sequence is identical for eight amino acids with the sequence of isoleucyl tRNA synthetase of E. coli. This finding adds to the evidence that mupirocin resistance is the result of a modified isoleucyl tRNA synthetase.
Insights
Mupirocin resistance in Staphylococcus aureus is linked to a modified isoleucyl tRNA synthetase. This study cloned and sequenced the resistance gene, confirming its role in altered enzyme function.
Area of Science:
- Microbiology
- Molecular Biology
- Genetics
Background:
- Staphylococcus aureus is a significant pathogen.
- Mupirocin is an important antibiotic for treating S. aureus infections.
- Antibiotic resistance, including to mupirocin, is a growing public health concern.
Purpose of the Study:
- To investigate the genetic basis of mupirocin resistance in Staphylococcus aureus.
- To identify and characterize the gene responsible for mupirocin resistance.
- To elucidate the molecular mechanism underlying mupirocin resistance.
Main Methods:
- Isolation and characterization of a mupirocin resistance-associated plasmid from Staphylococcus aureus.
- Construction of a restriction enzyme map of the plasmid.
- Cloning of a specific EcoRI fragment into an Escherichia coli-Staphylococcus aureus shuttle vector.
- DNA sequencing of the cloned fragment and comparison of derived amino acid sequence with databases.
Main Results:
- A 4.05 kb EcoRI fragment carrying the mupirocin resistance gene was successfully cloned.
- DNA sequencing revealed that the resistance gene encodes a modified isoleucyl tRNA synthetase.
- The derived amino acid sequence showed identity with E. coli isoleucyl tRNA synthetase.
Conclusions:
- The study provides strong evidence that mupirocin resistance in Staphylococcus aureus results from alterations in the isoleucyl tRNA synthetase enzyme.
- This finding contributes to understanding the molecular mechanisms of antibiotic resistance.
- The identified gene and its product are potential targets for future therapeutic strategies.