Table 1.
Plasmids and strains used in this study.
| Plasmids/Strains | Characteristics a | References |
|---|---|---|
| Plasmids | ||
| pMMB67EH | Broad host-range vector, Ptac, lacIq, AmpR | [30] |
| pKAS32 | Allele-exchange suicide vector, AmpR | [31] |
| pMMB67EH-PagLPa | pMMB67EH harboring pagL from P. aeruginosa PAO1 | [8] |
| pMMB67EH-LpxAPa | pMMB67EH harboring lpxA from P. aeruginosa PAO1 | This study |
| pMMB67EH-LpxLNm | pMMB67EH harboring lpxL1 from N. meningitidis H44/76 | This study |
| pMMB67EH-LpxLPg | pMMB67EH harboring lpxL from P. gingivalis ATCC33277 | This study |
| pMMB67EH-LpxDPa | pMMB67EH harboring lpxD from P. aeruginosa PAO1 | This study |
| pKA32-EFGH-lpxL2::gm | pKAS32 derivative harboring DNA segments of lpxL1 and downstream of lpxL2 separated by a gm cassette, AmpR, GmR | Geurtsen J |
| pKOlpxL2::gm (a) | pKA32-EFGH-lpxL2::gm derivative harboring a DNA segment upstream of lpxL1 and the entire lpxL2 gene except for the first 6 nucleotides together with a partial dap gene with, in between a gm cassette in the same orientation as the operon, AmpR, GmR | This study |
| pKOlpxL2::gm (b) | Like pKOlpxL2::gm (a) but with the gm cassette in the opposite orientation, AmpR, GmR | This study |
| pRTP113368K2a | pKAS32 derivative harboring DNA segments of lpxL2, AmpR, KanR (kan in similar orientation as the operon) | Hamstra HJ |
| pRTP113368K1a | pRTP113368K2a derivative harboring kan in opposite orientation as the operon. | Hamstra HJ |
| pRT669 | lpxA knockout construct, AmpR, KanR (kan in opposite orientation as the lpxA gene) | Hamstra HJ |
| pSS1129 | Suicide vector for allelic exchange, GmR, AmpR | [32] |
| pClpxAPa | pSS1129 derivative for replacement of lpxA gene of B. pertussis by lpxA gene of P. aeruginosa | This study |
| pClpxDPa | Construct for replacement of lpxD gene of B. pertussis by lpxA gene of P. aeruginosa | This study |
| Strains | ||
| Escherichia coli | ||
| DH5α | F−, Δ(lacZYA-argF)U169 thi-1 hsdR17 gyrA96 recA 1 endA 1 supE44 relA1 phoA Φ80 dlacZΔM15 | UU lab collection |
| SM10λpir | thi thr leu fhuA lacY supE recA::RP4–2-Tc::Mu λpir R6K KanR | [33] |
| BL21(DE3) | Contains gene for T7 DNA polymerase | Invitrogen |
| BL21-pLpxAPa | BL21(DE3) carrying pMMB67EH-LpxAPa | This study |
| BL21-pLpxLNm | BL21(DE3) carrying pMMB67EH-LpxLNm | This study |
| BL21-pLpxLPg | BL21(DE3) carrying pMMB67EH-LpxLPg | This study |
| BL21-pLpxDPa | BL21(DE3) carrying pMMB67EH-LpxDPa | This study |
| Bordetella pertussis | ||
| B213 | StrR derivative of strain Tohama I, NalR | [27] |
| B213-pLpxAPa | B213 carrying pMMB67EH-LpxAPa | This study |
| B213 ΔlpxA-pLpxAPa | B213 with inactivated lpxA gene carrying pMMB67EH-LpxAPa | This study |
| B213-pLpxLNm | B213 carrying pMMB67EH-LpxLNm | This study |
| B213-pLpxLPg | B213 carrying pMMB67EH-LpxLPg | This study |
| B213-pLpxDPa | B213 carrying pMMB67EH-LpxDPa | This study |
| B1917 | [28] | |
| B1917 lpxAPa | B1917 with lpxA gene replaced by lpxA from P. aeruginosa | This study |
| B1917 lpxDPa | B1917 with lpxD gene replaced by lpxD from P. aeruginosa | This study |
a Ptac, tac promoter; AmpR, ampicillin-resistant; GmR, gentamicin resistant, KanR, kanamycin-resistant; NalR, Nalidixic acid-resistant; StrR, streptomycin-resistant.