TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Description | Reference or source |
|---|---|---|
| E. coli strains | ||
| BL21 Gold(DE3) | Protein expression cells | Stratagene |
| K-12 ATCC 23716 | Parental strain | Laboratory stock |
| LMS4209 | K-12 (ATCC 23716) ytfE::Cmr | 25 |
| S. aureus strains | ||
| NCTC 8325 | Parental strain | Laboratory stock |
| RN4220 | Restriction-negative derivative of NCTC 8325; transformable by electroporation | Laboratory stock |
| LMSA0229 | RN4220 scdA::Ermr | This study |
| N. gonorrhoeae strains | ||
| F62 | Parental strain | Laboratory stock |
| JCGC704 | dnrN::Kanr | This study |
| JCGC212 | kat::ermC | 51 |
| JCGC705 | kat::ermC dnrN::Kanr | This study |
| Plasmids | ||
| pSP64D-E | Cloning vector carrying an erythromycin resistance cassette | 8 |
| pSPScdA | Upstream region and 5′ end of scdA cloned into pSP64D-E, next to the erythromycin resistance cassette | This study |
| pSUB11 | Epitope tagging plasmid carrying three copies of the FLAG tag and a kanamycin resistance cassette | 55 |
| pGEM-T Easy | Cloning vector | Promega |
| pGEMDnrN | Sequences upstream and downstream of the dnrN gene (NGO0653) cloned into pGEM-T Easy | This study |
| pGEMDnrN-KO | Sequences upstream and downstream of the dnrN gene (NGO0653) flanking a kanamycin resistance cassette cloned into pGEM-T Easy | This study |
| pET28a(+) | T7-based expression vector that inserts a sequence encoding a His6 tag at the N terminus | Novagen |
| pET-ScdA | S. aureus scdA gene cloned into pET28a(+) | This study |
| pGS57 | Fumarase A-expressing plasmid | 57 |
| pUC18 | Cloning vector | Laboratory stock |
| pScdA | pUC18 carrying the scdA gene of S. aureus and its promoter region | This study |
| pYtfE | pUC18 carrying the ytfE gene of E. coli and its promoter region | 24 |
| pRKISC | Plasmid for expression of the E. coli isc operon | 51 |
| pRKSUF | Plasmid for expression of the E. coli suf operon | 52 |