Table 1.

Bacterial strains and plasmids

Strain or plasmidDerivation and propertiesaReference(s)
Strains
 A66Type 3 parent strain; Cps+4
 AM161pJD330 × A66, insertion-duplication mutation downstream of cps3S; Emr Cps+This study
 AM179pCV182 × A66, Δcps3UMtnpA1; Cps+This study
 AM188A66 cps3D5(V283E); Cpsr (20%)This study
 AM199pJD369 × A66, insertion-duplication mutation downstream of cps3D (polar oncps3S); Emr CpsThis study
 AM201AM188 with repaired cps3D5 mutation; Cps+This study
 AM1000D39 Δcps2ABCDETFGHI; CpsThis study
 D39Type 2 parent strain; Cps+4
 JD600WU2; Strr Cps+20
 JD614JD600 cps3D3 (mutation located between bp 885 and 1802); Strr Cpsr (<6%)19, 20
 JD692JD600 cps3D4 (mutation located between bp 1027 and 1802); Strr Cpsr (<0.6%)19, 20
 JD770pJD330 × WU2, insertion-duplication mutation downstream of cps3S; Emr Cps+19, 20
 JD908pJD369 × WU2, insertion-duplication mutation downstream of cps3D (polar on cps3S); EmrCps19, 20
 JY1060WU2pgm-1(K381T); Cpsr (20%) PGMr26
 R36AD39 serially passaged 36 times in rabbit anti-type 2 serum, Δcps2ABCDETFGHI; Cps4, 31
 WU2Type 3 parent strain; Cps+10
Plasmids
 pJY4163 and pJY4164Cloning vectors for insertion-duplication mutagenesis; lack origin of replication for S. pneumoniae; multiple cloning site in opposite orientations in pJY4163 and pJY4164; Emr58
 pCV182pJY4164::3.14-kb [Sau3AI-PstI (‘cps3DSU’),SacI-HindIII (‘tnpA-plpA’)]; Emr27; this study
 pJD330pJY4163::2.38-kb Sau3AI (‘cps3DSU’); Emr20
 pJD369pJY4164::0.575-kbPvuII-RsaI (‘cps3DS’); Emr20
 pJD380pJY4164::0.356-kbSau3AI-SspI fragment from pJD330 (‘cps3D’); Emr20
  • a Emr, erythromycin resistant; Strr, streptomycin resistant; Cps+, encapsulated; Cps, nonencapsulated; Cpsr(percentage of parental level), reduced capsule production; PGMr, decreased PGM activity.