Construction of Staphylococcus aureus Proline Transport Gene Recombinant Plasmids

The bacterial species Staphylococcus aureus is responsible for a significant amount of infections and disease in humans each year. The emergence of antibiotic resistance within the species is of great concern. A recent study has suggested that proline transport is important for S. aureus survival in mice. This previous study specifically examined the putP gene, which codes for a high affinity proline transporter that brings the essential amino acid proline into the bacterial cell. The aims of this study were: 1) to construct a plasmid that replicates in S. aureus, containing a full-length copy of the putP gene, that will restore a mutation in the putP gene and 2) to construct a plasmid for creation of a stable deletion mutation in the putP gene of S. aureus strain RN6390. A replicating plasmid containing the full-length putP gene was constructed and it is in the process of being moved into S. aureus strain RN4220 putPcells. The plasmid to be used for generating the deletion mutation has also been constructed and is in the process of being moved into S. aureus strain RN4220.