Cloning, sequencing, and expression of the DNA gyrase genes from Staphylococcus aureus

Abstract

We have isolated and cloned the gyrA and gyrB genes from Staphylococcus aureus. These adjacent genes encode the subunits of DNA gyrase. The nucleotide sequence of a 5.9-kb region which includes part of an upstream recF gene, the whole of gyrB and gyrA, and about 1 kb of unknown downstream sequence has been determined. The gyrB and gyrA gene sequences predict proteins of 886 and 644 amino acid residues, respectively, which have significant homologies with the gyrase subunits of Escherichia coli and Bacillus subtilis. Residues thought to be important to the structure and function of the subunits are conserved. These genes have been expressed separately by using a T7 promoter vector. N-terminal sequencing of the cloned gene products suggests that the mature GyrB subunit exists mainly with its initial five residues removed. Protein sequencing also supports the interpretation of our DNA sequencing data, which are inconsistent in several placed with the recently published sequence of the same genes (E. E. C. Margerrison, R. Hopewell, and L. M. Fisher, J. Bacteriol. 174:1596-1603, 1992).