TABLE 1.
Strains and plasmids
Strain or plasmid | Genotype or relevant characteristics | Source or reference |
---|---|---|
Strains | ||
CC118 | araD139 Δ(ara leu)7697 ΔlacZ74 phoAΔ20 galE galK thi rpsE rpoB argE(Am) recA1 | 17 |
LT2 | S. typhimurium (wild-type) | |
DM271 | apbE42::Tn10d(Tc) | |
Plasmids | ||
pSU19 | Intermediate-copy-number cloning vector; Cmr | 2 |
pApbE1 | apbE, 1.15-kb insert in pSU19 that encodes ApbE; Cmr | |
pET20b | Overexpression vector containing T7-dependent promoter and cloning site for C-terminal His6 fusions; Ampr | Novagen |
pApbE9 | apbE with NdeI start site, no lipoprotein signal peptide sequence, inserted into SmaI site of pSU19; Cmr | |
pApbE10 | Encodes ApbEcyto; apbE without the lipoprotein signal peptide sequence under the expression of a T7 promoter, NdeI-HindIII fragment from pApbE9 ligated into pET20b; Ampr | |
pApbE11 | apbE with NdeI start site inserted into SmaI site of pSU19; Cmr | |
pApbE12 | Encodes ApbE; apbE under the expression of a T7 promoter, NdeI-HindIII fragment from pApbE11 ligated into pET20b; Ampr | |
pApbE(D21S) | apbE(D21S), 1.15-kb insert in pSU19 that encodes ApbE(D21S); Cmr | |
pT7-5 | Overexpression vector containing T7-dependent promoter; Ampr | |
pApbE3 | EcoRI-HindIII-fragment from pApbE1 containing apbE ligated into pT7-5; Ampr | |
pOmpF | 73-bp fragment encoding the signal peptide of ompF with NdeI and PstI sites; cloned into the PstI site of pSU19; Cmr | |
pApbE(C20A) | 900-bp fragment cloned into SmaI site of pSU19; Cmr fragment encodes apbE amino acids 21 to 350 with a C20A change | |
pApbEperi | Encodes ApbEperi; ompF-apbE(C20A) gene fusion expressed by the T7 promoter of pT7-7; Ampr |