Table 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Descriptiona | Source or reference |
|---|---|---|
| Strains | ||
| E. coli | ||
| DH5α | hsdR17 recA1 gyrA endA1 relA1 | Invitrogen |
| Rosetta 2(DE3) | F−ompT hsdSB(rB− mB−) gal dcm (DE3) pRARE2 (Camr) | Novagen |
| S. pyogenes | ||
| NZ131 | M49 serotype | D.R. Martin, New Zealand |
| rgg mutant | NZ131 rgg mutant; Emr | 11 |
| SA5 | NZ131 rgg mutant complemented with pSA3; Emr Kanr; expresses an Rgg-Myc fusion protein | This study |
| wt::luc strain | NZ131 transformed with pKSM720; Specr | This study |
| wt::PspeB-luc strain | NZ131 transformed with pSA11; Specr | This study |
| wt::Pspd3-luc strain | NZ131 transformed with pSA12; Specr | This study |
| wt::PpulA-luc strain | NZ131 transformed with pSA13; Specr | This study |
| wt::P396-luc strain | NZ131 transformed with pSA14; Specr | This study |
| wt::P746-luc strain | NZ131 transformed with pSA15; Specr | This study |
| wt::P1113-luc strain | NZ131 transformed with pSA16; Specr | This study |
| rgg::luc strain | NZ131 rgg mutant transformed with pKSM720; Specr | This study |
| rgg::PspeB-luc strain | NZ131 rgg mutant transformed with pSA11; Specr | This study |
| rgg::Pspd3-luc strain | NZ131 rgg mutant transformed with pSA12; Specr | This study |
| rgg::PpulA-luc strain | NZ131 rgg mutant transformed with pSA13; Specr | This study |
| rgg::P396-luc strain | NZ131 rgg mutant transformed with pSA14; Specr | This study |
| rgg::P746-luc strain | NZ131 rgg mutant transformed with pSA15; Specr | This study |
| rgg::P1113-luc strain | NZ131 rgg mutant transformed with pSA16; Specr | This study |
| Plasmids | ||
| pTrcHis2B | Cloning vector; Ampr | Invitrogen |
| pSA1 | rgg ORF cloned into pTrcHis2B; Ampr | This study |
| pKK1 | Shuttle vector defective for replication in S. pyogenes; Kanr | 26 |
| pSA3 | rgg ORF in frame with Myc and 6× histidine tags cloned into pKK; Kanr | This study |
| pMAL-c4X | Cloning vector; Ampr | New England BioLabs |
| pSA4 | rgg ORF cloned into pMAL-c4X; Ampr | This study |
| pSA5 | Promoter region of speB cloned into pGEM-T-vector; Ampr | This study |
| pSA6 | Promoter region of spd3 cloned into pGEM-T-vector; Ampr | This study |
| pSA7 | Promoter region of pulA cloned into pGEM-T-vector; Ampr | This study |
| pSA8 | Promoter region of Spy49_0396 cloned into pGEM-T-vector; Ampr | This study |
| pSA9 | Promoter region of Spy49_0746 cloned into pGEM-T-vector; Ampr | This study |
| pSA10 | Promoter region of Spy49_1113 cloned into pGEM-T-vector; Ampr | This study |
| pKSM720 | GAS replicating plasmid with firefly luciferase and RBS; Specr | 28 |
| pSA11 | Noncoding region upstream of speB cloned into pKSM720; Specr | This study |
| pSA12 | Noncoding region upstream of spd3 cloned into pKSM720; Specr | This study |
| pSA13 | Noncoding region upstream of pulA cloned into pKSM720; Specr | This study |
| pSA14 | Noncoding region upstream of Spy49_0396 cloned into pKSM720; Specr | This study |
| pSA15 | Noncoding region upstream of Spy49_0746 cloned into pKSM720; Specr | This study |
| pSA16 | Noncoding region upstream of Spy49_1113 cloned into pKSM720; Specr | This study |
a
RBS, ribosome binding site.