TABLE 1.

Strains used in the studya

StrainGenotype (derivation)Antibiotic resistancebReference or source
EL59Unencapsulated laboratory strain R6 derived from intermediates of strain D39None33
EL1454R6 kant1t2-Pc-pcsB+Kanr46
IU1533R6 ΔbgaA′::kant1t2-Pc-pcsB+Kanr45
IU1545R6 kant1t2-PfcsK-pcsB+Kanr45
IU1547R6 ΔpcsB<>Pc-ermAM ΔbgaA<> kant1t2-Pc-pcsBErmr Kanr45
IU1614E. coli BL21(DE3) [pET22B::pcsB(ΔN27)]AmprThis study
IU1621R6 kant1t2-Pc-pcsB+ ΔbgaA′::Pc-aad9 (new amplicon × IU1454)Kanr SpecrThis study
IU1690Serotype 2 encapsulated D39 (NCTC 7466)None33
IU1705R6 ΔdacA::Pc-aad9 (new amplicon × EL59)SpecrThis study
IU1744R6 kant1t2-Pc-pcsB+ (amplicon from EL1454 × EL59)KanrThis study and reference 45
IU1751R6 ΔmreCD<>aad9 (new amplicon × EL59)SpecrThis study
IU1781D39 rpsL1Strr53
IU1783D39 rpsL1 Δcps2BCDE<>kan-rpsL (new amplicon × IU1781)Strr Kanr33
IU1807D39 kant1t2-Pc-pcsB+ (amplicon from EL1454 × IU1690)KanrThis study
IU1845R6 pcsB+-FLAG Pc-ermAM (new amplicon × EL59)ErmrThis study
IU1859E. coli BL21(DE3) [pET15B::mreC(ΔN32)]AmprThis study
IU1945cD39 Δcps2A′ (cps2BCDETFG)H′None33
IU1979R6 ΔpcsB<>ermAM ΔbgaA′::kant1t2-Pc-pcsB+ (new amplicon × IU1533)Ermr KanrThis study
IU1983R6 ΔmreCD<>aad9 (amplicon from IU1751 × EL59)SpecrThis study
IU2091D39 kant1t2-PfcsK-pcsB+ (amplicon from IU1545 × IU1690)KanrThis study
IU2102D39 ΔmreCD<>aad9 (amplicon from IU1751 × IU1690)SpecrThis study
IU2105D39 pcsB+-FLAG Pc-ermAM (amplicon from IU1845 × IU1690)ErmrThis study
IU2142L. lactis NZ9000 (pLS1RGFP) (plasmid × NZ9000)Ermr65
IU2191R6 (pLS1RGFP) (plasmid × EL59)ErmrThis study
IU2192R6 (pLS1RGFP::Pmal-pcsB+) (ligation mixture × EL59)ErmrThis study
IU2336D39 Δcps2A′ (cps2BCDETFG)H′ kant1t2-Pc-pcsB+ (amplicon from IU1744 × IU1945)KanrThis study
IU2517D39 ΔbgaA′::kant1t2-Pc-pcsB+ (amplicon from IU1533 × IU1690)KanrThis study
IU2519D39 ΔbgaA′::kant1t2-PfcsK-pcsB+ (new amplicon × IU1690)KanrThis study
IU2537D39 ΔpcsB<>ermAM ΔbgaA′::kant1t2-Pc-pcsB+ (amplicon from IU1979 × IU2517)Ermr KanrThis study
IU2539D39 ΔpcsB<>ermAM ΔbgaA′::kant1t2-PfcsK-pcsB+ (amplicon from IU1979 × IU2519)Ermr KanrThis study
IU2547D39 ΔbgaA′::Pc-aad9 (amplicon from IU1621 × IU1690)SpecrThis study
IU2551D39 Δcps2A′ (cps2BCDETFG)H′ kant1t2-Pc-pcsB+ (pLS1RGFP::Pmal-pcsB+) (plasmid from IU2192 × IU2336)Kanr ErmrThis study
IU2564R6 kant1t2-Pc-pcsB+ (pLS1RGFP::Pmal-pcsB+) (plasmid from IU2192 × IU1744)Kanr ErmrThis study
IU2566D39 Δcps2A′ (cps2BCDETFG)H′ (pLS1RGFP::Pmal-pcsB+) (plasmid from IU2192 × IU1945)ErmrThis study
IU2592R6 ΔbgaA′::kant1t2-PfcsK-pcsB+ (amplicon from IU2519 × EL59)KanrThis study
IU2604D39 Δcps2A′ (cps2BCDETFG)H′ (pLS1RGFP) (plasmid from IU2191 × IU1945)ErmrThis study
IU2608R6 kant1t2-Pc-pcsB+ (pLS1RGFP) (plasmid from IU2191 × IU1744)Kanr ErmrThis study
IU2610D39 Δcps2A′ (cps2BCDETFG)H′ kant1t2-Pc-pcsB+ (pLS1RGFP) (plasmid from IU2191 × IU2336)Kanr ErmrThis study
IU2718R6 ΔbgaA′::ermt1t2-PfcsK-pcsB+ (A27D) (new amplicon x EL59)ErmrThis study
IU2801R6 ΔpcsB<>ermAM ΔbgaA::kant1t2-PfcsK-pcsB+ (amplicon from IU1979 × IU2592)Ermr KanrThis study
IU2824D39 ΔdacA::Pc-aad9 (amplicon from IU1705 × IU1690)SpecrThis study
IU2825D39 Δcps2A′ (cps2BCDETFG)H′ ΔdacA::Pc-aad9 (amplicon from IU1705 × IU1945)SpecrThis study
IU3192D39 Δcps2BCDE<>kan-rpsL (amplicon from IU1783 × IU1690)KanrThis study
  • a Strains are derivatives of S. pneumoniae EL59 (R6) and IU1690 (D39), unless indicated otherwise. Strains were constructed by transformation (indicated by ×) of indicated recipients with PCR fragments amplified from the chromosomes of other strains or synthesized by fusion PCR (see Materials and Methods). <> or :: indicates exact replacement of a reading frame or insertion, respectively.

  • b Antibiotic resistance markers: Specr, spectinomycin; Ermr, erythromycin; Kanr, kanamycin; Strr, streptomycin; Ampr, ampicillin (in E. coli strains only).

  • c IU1945 was constructed to have the same Δcps region as EL59 (R6) (33).