Table 2.
Strains used in this studya
| Strain | Genotype | Derivation | Antibiotic resistanceb | Source(s) |
|---|---|---|---|---|
| EL59 | Unencapsulated laboratory strain R6 | Derived from intermediates of strain D39 | None | 28 |
| EL556 | R6 ΔlytA<>Pc-aad9 | ΔlytA<>Pc-aad9 × R6 | Spec | 57 |
| IU1690 | Serotype 2 capsulated D39 | NTCC7466 | None | 28 |
| IU1708 | R6 ΔcbpD<>Pc-kan-rpsL | ΔcbpD<>Pc-kan-rpsL × EL59 | Kan | This study |
| IU1732 | R6 Δspd_1874::aad9 (3′ 27 ntd of spd_1874 remain) | Δspd_1874::aad9 × EL59 | Spec | This study |
| IU1945 | D39 Δcps2A′(cps2BCDETFG)H′ | Δcps2A′(cps2BCDETFG)H′ × IU1690 | None | 28 |
| IU2824 | D39 ΔdacA<>Pc-aad9 | ΔdacA<>Pc-aad9 × IU1690 | Spec | 1 |
| IU2825 | D39 Δcps2A′(cps2BCDETFG)H′ ΔdacA<>Pc-aad9 | ΔdacA<>Pc-aad9 × IU1945 | Spec | 1 |
| IU3766 | D39 ΔlytA<>Pc-aad9 | ΔlytA<>Pc-aad9 × IU1690 (amplicon from EL556) | Spec | This study, 57 |
| IU3775 | D39 ΔcbpD<>Pc-kan-rpsL | ΔcbpD<>Pc-kan-rpsL × IU1690 (amplicon from IU1708) | Kan | This study |
| IU3795 | D39 Δspd_1874::aad9 (3′ 27 nt of spd_1874 remain) | Δspd_1874::aad9 × IU1690 (amplicon from IU1732) | Spec | This study |
| IU3797 | D39 Δpmp23<>Pc-kan | Δpmp23<>Pc-kan × IU1690 | Kan | This study |
| IU3800 | D39 ΔlytC<>Pc-kan-rpsL | ΔlytC<>Pc-kan-rpsL × IU1690 | Kan | This study |
| IU3801 | D39 Δspd_0873<>Pc-kan-rpsL | Δspd_0873<>Pc-kan-rpsL × IU1690 | Kan | This study |
| IU3804 | D39 Δspd_0173<>Pc-kan-rpsL | Δspd_0173<>Pc-kan-rpsL × IU1690 | Kan | This study |
| IU3805 | D39 ΔdacB<>Pc-kan-rpsL | ΔdacB<>Pc-kan-rpsL × IU1690 | Kan | This study |
| IU3806 | D39 Δspd_0703<>kan | Δspd_0703<>kan × IU1690 | Kan | This study |
| IU3837 | D39 ΔlytB<>Pc-kan-rpsL | ΔlytB<>Pc-kan-rpsL × IU1690 | Kan | This study |
| IU3874 | D39 Δcps2A′(cps2BCDETFG)H′ ΔcbpD<>Pc-kan-rpsL | ΔcbpD<>Pc-kan-rpsL × IU1945 (amplicon from IU1708) | Kan | This study |
| IU3875 | D39 Δcps2A′(cps2BCDETFG)H′ Δpmp23<>Pc-kan | Δpmp23<>Pc-kan × IU1945 | Kan | This study |
| IU3876 | D39 Δcps2A′(cps2BCDETFG)H′ ΔlytC<>Pc-kan-rpsL | ΔlytC<>Pc-kan-rpsL × IU1945 | Kan | This study |
| IU3877 | D39 Δcps2A′(cps2BCDETFG)H′ ΔlytB<>Pc-kan-rpsL | ΔlytB<>Pc-kan-rpsL × IU1945 | Kan | This study |
| IU3878 | D39 Δcps2A′(cps2BCDETFG)H′ Δspd_0873<>Pc-kan-rpsL | Δspd_0873<>Pc-kan-rpsL × IU1945 | Kan | This study |
| IU3879 | D39 Δcps2A′(cps2BCDETFG)H′ Δspd_0173<>Pc-kan-rpsL | Δspd_0173<>Pc-kan-rpsL × IU1945 | Kan | This study |
| IU3880 | D39 Δcps2A′(cps2BCDETFG)H′ ΔdacB<>Pc-kan-rpsL | ΔdacB<>Pc-kan-rpsL × IU1945 | Kan | This study |
| IU3881 | D39 Δcps2A′(cps2BCDETFG)H′ Δspd_0703<>kan | Δspd_0703<>kan × IU1945 | Kan | This study |
| IU3900 | D39 Δcps2A′(cps2BCDETFG)H′ ΔlytA<>Pc-aad9 | ΔlytA<>Pc-aad9 × IU1945 (amplicon from EL556) | Spec | This study, 57 |
| IU3901 | D39 Δcps2A′(cps2BCDETFG)H′ Δspd_1874::aad9 (3′ 27 nt of spd_1874 remain) | Δspd_1874::aad9 × IU1945 (amplicon from IU1732) | Spec | This study |
| IU3917 | D39 Δcps2A′(cps2BCDETFG)H′ Δspd_0104<>Pc-kan-rpsL (5′ 30 nt and 3′ 30 nt of spd_0104 remain) | Δspd_0104<>Pc-kan-rpsL × IU1945 | Kan | This study |
| IU3957 | D39 Δcps2A′(cps2BCDETFG)H′ ΔdacA<>Pc-aad9 ΔdacB<>Pc-kan-rpsL | ΔdacB<>Pc-kan-rpsL × IU2825 | Spec, Kan | This study |
| IU4054 | D39 rpsL1 Δspd_0104::Pc-kan-rpsL (5′ 30 nt and 3′ 30 nt of spd_0104 remain) | Δspd_0104<>Pc-kan-rpsL × IU1690 | Kan | This study |
| IU4259 | D39 ΔdacB<>Pc-kan-rpsL bgaA::Pc-ermAM-PfcsK-dacB | bgaA::Pc-ermAM-PfcsK-dacB × IU3805 | Kan, Erm | This study |
| IU4787 | D39 Δspd_0703<>kan bgaA::Pc-ermAM-PfcsK-spd_0703 | bgaA::Pc-ermAM-PfcsK-spd_0703 × IU3806 | Kan, Erm | This study |
| IU4960c | D39 Δcps2A′(cps2BCDETFG)H′ dacA-L-FFF | dacA+-L-FFF-Pc-aad9 × IU1945 | Spec | This study |
| IU4961c | D39 Δcps2A′(cps2BCDETFG)H′ dacB-L-FFF | dacB+-L-FFF-Pc-kan × IU1945 | Kan | This study |
a
The genetic background was capsulated serotype 2 strain D39 (IU1690) or its isogenic unencapsulated derivative IU1945. Strains were constructed by transformation of amplicons (left of × sign) into the indicated recipient strain (right of × sign) as described in Materials and Methods. The symbols <> and :: indicate replacement of and insertion into a reading frame with an antibiotic marker, respectively.
b
Antibiotic resistance markers: Erm, erythromycin; Kan, kanamycin; Spec, spectinomycin.
c
L-FFF refers to a carboxyl-terminal addition of a 10-amino-acid spacer/linker (GSAGSAAGSG) followed by three tandem copies of the FLAG epitope (DYKDDDDK) (see reference 69).
d
nt, nucleotides.