Table 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Descriptiona | Source or reference |
|---|---|---|
| S. pneumoniae strains | ||
| D39 | S. pneumoniae serotype 2 | 63 |
| ST581 | D39 with rpsL1 allele; Strepr | 42 |
| ST2718 | Δpde1::Janus cassette in ST581; Kanr | This study |
| ST2719 | Δpde2::Janus cassette in ST581; Kanr | This study |
| ST2729 | In-frame deletion mutant of pde1 in ST581; Strepr | This study |
| ST2730 | In-frame deletion of pde1 and Δpde2::Janus cassette; Kanr | This study |
| ST2733 | In-frame deletion mutant of pde2 in ST581; Strepr | This study |
| ST2734 | In-frame deletion mutant of pde1 and pde2 in ST581; Strepr | This study |
| ST2847 | ST581(pVA838); Strepr Ermr | This study |
| ST2848 | ST2733(pST2846); Strepr Ermr | This study |
| ST2849 | ST2733(pVA838); Strepr Ermr | This study |
| ST2850 | ST2729(pST2846); Strepr Ermr | This study |
| ST2851 | ST2729(pVA838); Strepr Ermr | This study |
| E. coli strains | ||
| DH5α | E. coli strain used for cloning | Laboratory stock |
| BL21(DE3) | E. coli strain used for expression | Novagen |
| ST2714 | E. coli BL21(DE3)(pST2710); Kanr | This study |
| ST2715 | E. coli BL21(DE3)(pST2711); Kanr | This study |
| ST2761 | E. coli DH5α(pST2761); Apr | This study |
| ST2762 | E. coli DH5α(pST2762); Apr | This study |
| Plasmids | ||
| pET28a(+) | His tag expression vector; Kanr | Novagen |
| pProEXHTb | His tag expression vector; Apr | Invitrogen |
| pVA838 | E. coli-S. pneumoniae shuttle vector; Ermr | 64 |
| pST2710 | pET28a(+) carrying S. pneumoniae dacA ORF; Kanr | This study |
| pST2711 | pET28a(+) carrying S. pneumoniae pde2 ORF; Kanr | This study |
| pST2761 | pProEX HTb carrying S. pneumoniae pde1 aa 51–657 ORF; Apr | This study |
| pST2762 | pProEX HTb carrying S. pneumoniae pde1 aa 109–657 ORF; Apr | This study |
| pST2846 | pVA838 carrying S. pneumoniae pde2 promoter and ORF; Ermr | This study |
a
Strepr, streptomycin resistance; Kanr, kanamycin resistance; Ermr, erythromycin resistance; Apr, ampicillin resistance.