TABLE 1.
Bacterial strains and plasmids
| Strain or plasmid | Derivationa | Reference or source |
|---|---|---|
| S. pneumoniae strains | ||
| GA16638 | Parent type 14, type 2 mega, Emr | This study |
| GA17719 | Parent type 14, type 1 mega, Emr | This study |
| KA3000/01 | pKA309 × GA16638 ΔmefE Ems | This study |
| KA3003/04 | pKA312 × GA16638 Δmel Ems | This study |
| KA3005/06 | pKA321 × GA16638 ΔmefE Δmel Ems | This study |
| ATCC 49619 | 26 | |
| E. coli DH5α plasmids | ||
| pCR2.1-TOPO | Vector for cloning of PCR products, Apr Knr | Invitrogen |
| pSF151 | 41 | |
| pWA101 | TOP10 pCR2.1::0.34-kb KG7/KG11 PCR product from GA16638 | This study |
| pWA103 | TOP10 pCR2.1::0.56-kb KG8/KG10 PCR product from GA16638 | This study |
| pKA309 | DH5α pSF151::double-fragment ligation of KpnI/XbaI insert of pWA101 and KpnI/PstI insert of pWA103 ligated to XbaI/PstI of vector | This study |
| pKA310 | TOP10 pCR2.1::0.66-kb KG20/KG41R PCR fragment from GA16638 | This study |
| pKA312 | DH5α pSF151::double-fragment ligation of SacI/EcoRV fragment from pWA103 ligated to EcoRV/SpeI fragment of pKA310 | This study |
| pKA321 | DH5α pSF151::0.34-kb EcoRI/SpeI fragment of pWA101 and 0.6-kb XbaI/BamHI fragment of pKA310 ligated to EcoRI/BamHI of vector, Knr | This study |
a
Emr, erythromycin resistant; Ems, erythromycin sensitive; Apr, ampicillin resistant; Knr, kanamycin resistant. × indicates transformation (or crossing) of the plasmid into the strain. Primers are listed in Table 2.