TABLE 2.
Plasmids used in this study
| Plasmid | Relevant characteristic(s) | Reference or source |
|---|---|---|
| pBluescriptII KS(−) | Blue/white cloning vector | Stratagene |
| pET21a | Six-His vector | Novagen |
| pMal-c2 | malE MBP fusion vector | New England Biolabs |
| pMga4-4 | WTmga4ain pLZ12-Spec | 2 |
| pMga50-His | WT mga50 gene with C-terminal six-His tag | This study |
| pPmga-blue | Pmga in pBluescript II KS(−) | This study |
| pPmga-gusA | gusA under Pmga6 | This study |
| pPmrp-gusA | gusA under Pmrp | This study |
| pVIT164 | GAS integration vector | 6 |
| pJRS312 | Spectinomycin cassette | 26 |
| pJRS515 | WT mga6 in pBluescript II KS(−) | 18 |
| pJRS547 | Deletion of 2.1 kb of mga and Pmga (mga-16) | This study |
| pJRS9160 | Counterselectable gene replacement vector | 20 |
| pKSM140 | Promoterless gusA in pCIV2 suicide vector | This study |
| pKSM155 | N-terminal MBP fusion to M4 Mga | This study |
| pKSM156 | N-terminal MBP fusion to M50 Mga | This study |
| pKSM162 | WT mga6 under constitutive Pspac | 20 |
| pKSM164 | WT mga6-his under native Pmga6 | 1 |
| pKSM174 | mga-16 deletion allele in pJRS9160 | This study |
| pKSM315 | WT mga50 under constitutive Pspac | This study |
| pKSM318 | WT mga6-his under constitutive Pspac | This study |
| pKSM320 | WT mga4-his under constitutive Pspac | This study |
| pKSM321 | WT mga50-his under constitutive Pspac | This study |
| pKSM317-H | mga50-his (S26N) under constitutive Pspac | This study |
| pP361A-H | mga50-his (P361A) under constitutive Pspac | This study |
| pR461M-H | mga50-his (R461M) under constitutive Pspac | This study |
| pKSM322 | WT mga4-his under native Pmga4 | This study |
| pSKM323 | mga50-his (S26N) under native Pmga4 | This study |
| pKSM324 | WT mga4-his under constitutive PrpsL | This study |
| pKSM325 | WT mga50-his under constitutive PrpsL | This study |
| pKSM326 | mga50-his (S26N) under constitutive PrpsL | This study |
| pKSM327 | mga50-his (P361A) under constitutive PrpsL | This study |
| pKSM328 | mga50-his (R461M) under constitutive PrpsL | This study |
| pKSM329 | WT mga50-his under native Pmga4 | This study |
| pKSM330 | mga50-his (R461M) under native Pmga4 | This study |
| pKSM331 | mga50-his (S26N/R461M) under native Pmga4 | This study |
| pKSM332 | mga50-his (S26N/R461M) under constitutive PrpsL | This study |
| pKSM333 | WT mga6-his under constitutive PrpsL | This study |
| pKSM336 | mga50-his (P361A) under native Pmga4 | This study |
| pKSM337 | mga6-his (M461R) under constitutive Pmga6 | This study |
| pKSM338 | mga6-his (M461R) under constitutive PrpsL | This study |
a
WT, wild type.