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 (Chlr) | Novagen |
| S. pyogenes | ||
| NZ131 | M49 serotype | D. R. Martin, New Zealand |
| rgg mutant | NZ131 rgg mutant, Eryr | 15 |
| SA5 | NZ131 rgg mutant complemented with pSA3, Eryr Kanr | 2 |
| wt::luc | NZ131 transformed with pKSM720, Sptr | 2 |
| wt::Pemm49-luc | NZ131 transformed with pSA21, Sptr | This study |
| wt::Psof-luc | NZ131 transformed with pSA22, Sptr | This study |
| wt::PsfbX49luc | NZ131 transformed with pSA23, Sptr | This study |
| wt::PspeH-luc | NZ131 transformed with pSA24, Sptr | This study |
| rgg::luc | NZ131 rgg mutant transformed with pKSM720, Eryr Sptr | 2 |
| rgg::Pemm49-luc | NZ131 rgg mutant transformed with pSA21, Eryr Sptr | This study |
| rgg::Psof-luc | NZ131 rgg mutant transformed with pSA22, Eryr Sptr | This study |
| rgg::PsfbX49-luc | NZ131 rgg mutant transformed with pSA23, Eryr Sptr | This study |
| rgg::PspeH-luc | NZ131 rgg mutant transformed with pSA24, Eryr Sptr | This study |
| lacD.1 mutant | NZ131 lacD.1 mutant, Eryr | This study |
| SA6 | NZ131 lacD1 mutant complemented with pSA26, Eryr Kanr | This study |
| lacD.1::PspeH-luc | NZ131 lacD.1 mutant transformed with pSA24, Eryr Sptr | This study |
| SA6::PspeH-luc | NZ131 lacD.1 complemented strain (SA6) transformed with pSA24, Eryr Kanr Sptr | This study |
| lacD.1::Pemm49-luc | NZ131 lacD.1 mutant transformed with pSA21, Eryr Sptr | This study |
| SA6::Pemm49-luc | NZ131 lacD.1 complemented strain (SA6) transformed with pSA21, Eryr Kanr Sptr | This study |
| lacD.1::Psof-luc | NZ131 lacD.1 mutant transformed with pSA22, Eryr Sptr | This study |
| SA6::Psof-luc | NZ131 lacD.1 complemented strain (SA6) transformed with pSA22, Eryr Kanr Sptr | This study |
| lacD.1::PsfbX49-luc | NZ131 lacD.1 mutant transformed with pSA23, Eryr Sptr | This study |
| SA6::PsfbX49-luc | NZ131 lacD.1 complemented strain (SA6) transformed with pSA23, Eryr Kanr Sptr | This study |
| Plasmids | ||
| pVA891-2 | Suicide cloning vector, Eryr | 40 |
| pVaLac | Shuttle vector defective for replication in S. pyogenes; Eryr | This study |
| pSA17 | Promoter region of emm49 cloned into pGEM-T vector, Ampr | This study |
| pSA18 | Promoter region of sof cloned into pGEM-T vector, Ampr | This study |
| pSA19 | Promoter region of sfbX49 cloned into pGEM-T vector, Ampr | This study |
| pSA20 | Promoter region of speH cloned into pGEM-T vector, Ampr | This study |
| pKSM720 | GAS replicating plasmid with firefly luciferase and RBS, Sptr | 33 |
| pSA21 | Noncoding region upstream of emm49 cloned into pKSM720, Sptr | This study |
| pSA22 | Noncoding region upstream of sof cloned into pKSM720, Sptr | This study |
| pSA23 | Noncoding region upstream of sfbX49 cloned into pKSM720, Sptr | This study |
| pSA24 | Noncoding region upstream of speH cloned into pKSM720, Sptr | This study |
| pMNN23 | Streptococcal expression plasmid containing rofA promoter, Kanr | 45 |
| pSA26 | lacD.1 open reading frame cloned into pMNN23 | This study |
a
Chlr, chloramphenicol resistant; Eryr, erythromycin resistant; Kanr, kanamycin resistant; Sptr, spectinomycin resistant; Ampr, ampicillin resistant; GAS, group A streptococcus; RBS, ribosome-binding site.