TABLE 1.
Bacterial strains and plasmids
| Strain or plasmida | Derivation and propertiesb,c | Reference(s) |
|---|---|---|
| S. pneumoniae strains | ||
| D39 | Type 2 encapsulated; PspA+ (84 kDa) | 1, 33 |
| GH1003 | pGG108 × WU2; Emr; insertion downstream of pspA; PspA+ (92 kDa) | This study |
| JD611 | Nonencapsulated derivative of WU2; Cps3D−; PspA+ (92 kDa) | 10 |
| JD770 | Type 3 encapsulated derivative of WU2 containing insertion-duplication of cps3DSU; Emr; PspA+ (92 kDa) | 10 |
| JD803 | JD770 × D39; Emr; type 3 encapsulated; PspA+ (84 kDa) | 25 |
| JD804 | Same as JD803; independent derivative | 25 |
| JY1123 | pKSD300 × WU2; Emr; PspAtr (46 kDa) | 45 |
| MA101 | pJY4173 × D39; Emr; insertion downstream of pspA; PspA+ (84 kDa) | This study |
| MA139 | GH1003 × D39; Emr; PspA+ (92 kDa) | This study |
| MA158 | Same as MA139; independent derivative | This study |
| MA200 | WG44.1 × R36A; Emr; PspA− | This study |
| MA1024 | MA101 × WU2; Emr; PspA+ (84 kDa) | This study |
| MA1025 | Same as MA1024; independent derivative | This study |
| R36A | Nonencapsulated derivative of D39; PspA+ (84 kDa) | 1 |
| Rx1 | Avirulent derivative of D39; PspA+ (84 kDa) | 33, 39 |
| WG44.1 | PspA− derivative of Rx1 (ΔpspA); Emr | 36, 45 |
| WU2 | Type 3 encapsulated, virulent; PspA+ (92 kDa) | 3, 33 |
| Plasmids | ||
| pGG103 | pJY4163::1.8-kb Sau3AI-KpnI fragment containing 3′ region of Rx1/WU2 pspA | This study |
| pGG108 | pJY4163::1.2-kb SacI-KpnI fragment containing WU2 pspA 3′ region | This study |
| pGH160 | pUC18::1.2-kb SphI-SalI containing WU2 pspA 5′ region; encodes PspA46 | This study |
| pJY4163 | Vector for insertion-duplication mutagenesis; replicates in E. coli, not in S pneumoniae; Emr | 45 |
| pJY4173 | pJY4163 containing 3′ region of Rx1 pspA | 45 |
| pJY4306 | pUC18::1.0-kb HindIII-DraI fragment (Rx1 pspA′ bp 1-997); encodes PspA42 | 45 |
| pJY4310 | pUC18::HindIII-SacI (Rx1 pspA′ bp 1-790); encodes PspA30 | 45 |
| pKSD300 | pVA891::0.55-kb internal Sau3AI fragment of Rx1 pspA | 36, 43 |
| pRS102 | HindIII self-ligated fragment from JY1123 (in pVA891); C-terminal coding region of WU2 pspA; Emr | This study |
| pVA891 | Vector for insertion-duplication mutagenesis; replicates in E. coli, not in S. pneumoniae; Cmr (E. coli only); Emr | 29 |
a
Strains MA139, MA158, MA1024, MA1025, JD803, and JD804 are each the result of at least three backerosses with the parental recipients.
b
Crosses were done by transformation.
c
The apparent molecular masses of PspA (in parentheses) were determined by SDS-PAGE and Western immunoblotting. PspA+, full-length PpA; PspAtr, truncated PspA.