TABLE 1.
Bacterial strains and plasmids
| Strain or plasmid | Relevant characteristicsa | Source or reference |
|---|---|---|
| Strains | ||
| AM604 | MC4100 ara+ | 41 |
| AM661 | AM604 ΔlptD::kan/araBp-lptD | 29 |
| AM689 | AM604 ΔlptE::kan Δ(λatt-lom)::blaaraBp-lptE | 41 |
| DH5α | Δ(argF-lac169) 80dlacZ58(M15) glnV44(AS) λ−rfbD1 gyrA96 recA1 endA1 spoT1 thi-1 hsdR17 | 11 |
| FL905 | AM604 Φ(kan araC araBp-lptC)1 | 29 |
| FL907 | AM604 Φ(kan araC araBp-lptA)1 | 29 |
| M15/pREP4 | F−lac thi mtl/pREP4 | Qiagen |
| Plasmids | ||
| pRSET | T7 promoter; Apr | Invitrogen |
| pET30b | T7 promoter; Kmr | Novagen |
| pET30-lptC-lk25-H | pET30b derivative, expresses a LptC-lk25-His6 version where lk is a 25-amino-acid linker region separating LptC residue 191 from the His6 affinity tag | This study |
| pQE30 | T5 promoter; Apr | Qiagen |
| pQEsH-lptC | pQE30 derivative, expresses His6-LptC24-191 | This study |
| pGS100 | pGZ119EH derivative, contains TIR sequence downstream of Ptac; Cmr | 30 |
| pGS103 | pGS100 Ptac-lptC | 30 |
| pGS103Δ177-191 | pGS103 Ptac-lptCΔ177-191 | This study |
| pGS103G56V | pGS103 Ptac-lptC(G56V) | This study |
| pGS103Y112S-G153R | pGS103 Ptac-lptC(Y112S-G153R) | This study |
| pGS103G153R | pGS103 Ptac-lptC(G153R) | This study |
| pGS108 | pGS100 Ptac-lptC-H | 30 |
| pGS108Δ177-191 | pGS108 Ptac-lptCΔ177-191-H | This study |
| pGS108G153R | pGS108 Ptac-lptC(G153R)-H | This study |
| pGS108G56V | pGS108 Ptac-lptC(G56V)-H | This study |
| pGS116 | pGS103Δ177-191 derivative expressing both the truncated LptCΔ177-191 and LptC-lk25 from pET30- lptC-lk25-H plasmid | This study |
a
Cmr, chloramphenicol resistance; Apr, ampicillin resistance; Kmr, kanamycin resistance.