TABLE 1.
Strains and plasmids
| Strain or plasmid | Relevant genotype | Phenotype | Source or reference |
|---|---|---|---|
| Strain | |||
| E. coli | |||
| XL1-Blue | F′::Tn10 proA+B+ lacIqΔ(lacZ)M15/recA1 endA1 gyrA96 (Nalr) thi hsdR17(rK− mK+) supE44 relA1 lac | Stratagene | |
| BL21(DE3) | F−ompT gal [dcm] [lon] hsdSB(rB− mB−) (DE3) | Novagen | |
| Top10 | F−mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74 recA1 araD139 Δ(ara leu)7697 galU galK rpsL (StrR) endA1 nupG | Invitrogen | |
| S. pneumoniae | |||
| Cp1015 | RX derivative; str1 hexA | Smr | 30 |
| Cp1016 | Cp1015, but rif23 and stgF | Rifr, Stgr | 5 |
| R119 | R6 hex, but rif23 stgF str1 fusA nov1 opt12 gln tdr | Smr, Fusr, Rifr, Stgr, Novr, Optr | 46 |
| Cp1008 | Cp1015, but comA::spc | Spr | 30 |
| Cp1095 | Cp1015, but lytA::ery | Eryr | 47, 49 |
| Cp7000 | Cp1015, but ΔstkP::cat | Camr | 31 |
| Cp9000 | Cp1015, but stkP::aphA-3 | Kmr | 17 |
| Cp9090 | Cp1095, but stkP::aphA-3 | Kmr, Eryr | 17 |
| Cp7004 | Cp7000, but phpPD1 | Camr | This work |
| Cp9091 | Cp9090, but stkP+ | Kans | This work |
| Cp6600 | Cp1015, but comD(D299N) rif23 | Rifr | 14 |
| Cp6607 | Cp6600, but ΔstkP::cat | Rifr, Camr | This work |
| Plasmid | |||
| pCR2.1TOPO | Cloning vector | Ampr, Kmr | Invitrogen |
| pWSK29 | Cloning vector derived from pBluescript | Ampr | 51 |
| pET28b | His tag expression vector; Kanr, f1 ori | Kmr | Novagen |
| pBluescript ISK+/KS+ | Ampr, lacZ, f1± ori, ColE1 ori | Ampr | Stratagene |
| pPJ1 | pUC derivative containing a HincII fragment carrying the kanamycin resistance gene | Ampr, Kmr | 32 |
| pPHK29 | 2.96-kb EcoRI/SalI amplicon (primers PHL and PCR) from S. pneumoniae Cp1015 DNA containing phpP and stkP genes cloned into pWSK29 | Ampr | 17 |
| pPK29 | 2.3-kb EcoRI/SalI amplicon (primers PCL and PCR) from S. pneumoniae Cp1015 DNA containing stkP gene in pWSK29 | Ampr | This work |
| pEXstkP-N | 0.825-kb NdeI/EcoRI amplicon (primers STKP-F and STKP-RT) from pPHK29 containing fragment (kinase domain) of stkP gene inserted into pET28b | Kmr | This work |
| pEX-PhpP | 0.74-kb amplicon (phpP) inserted into pET28b | Kmr | 31 |
| pEX-PASTA | 0.9-kb NcoI/EcoRI amplicon (primers STKP-PF and STKP-R2) from pPHK29 containing stkP gene inserted into pET28b | Kmr | This work |
| pMUT1 | 2.6-kb amplicon of phpPD1 stkP+ | Kmr | This work |
| pMUT2 | 2.6-kb amplicon of phpPD2 stkP+ | Kmr | This work |
| pMUT3 | 2.6-kb amplicon of phpPD1 ΔstkP::cat | Kmr | This work |
| pMUT4 | 2.6-kb amplicon of phpPD2 ΔstkP::cat | Kmr | This work |