Table 1 Bacterial strains and plasmids
From: IcsA autotransporter passenger promotes increased fusion protein expression on the cell surface
Strain or plasmid' | Relevant characteristics | Reference or source |
|---|---|---|
E. coli K-12 | ||
DH5α | F- φ80lac ZΔM15 Δ(lac ZYA-arg F)U169 rec A1 end A1 hsd R17(rk-, mk+) pho Asup E44 thi-1 gyr A96 rel A1 λ- | Gibco-BRL |
KMRM1 | DH5α [pKMRM1]; AmpR | [29] |
MLRM3 | UT5600 [pBAD33]; CmR | This study |
MLRM4 | UT5600 [pMLRM1]; CmR | This study |
MLRM20 | UT5600 [pMLRM2::bla (Not I - Not I)], CmR | This study |
MLRM28 | UT5600 [pMLRM1::bla (Not I - Not I)], CmR | This study |
MLRM37 | UT5600 [pMLRM2::malE (Not I - Not I)], CmR | This study |
MLRM39 | UT5600 [pMLRM1::malE (Not I - Not I)], CmR | This study |
RMA3045 | E1315 [pMAL-c2], AmpR | Laboratory collection |
UT5600 | F- ara-14 leuB6 secA6 lacY1 proC14 tsx-67 Δ(ompT-fepC)266 entA403 trpE38 rfbD1 rpsL109 xyl-5 mtl-1 thi-1 | Laboratory collection |
Plasmids | ||
pBAD33 | pBAD expression; pACYC184 ori; CmR | [30] |
pIcsA | icsA (EcoR I - Sal I) gene cloned into pBR322; medium copy no.; ColE1 ori; AmpR | [33] |
pKMRM1 | pIcsA::5aa insertion at amino acid position 87 (icsA i87 or icsA) (bla); AmpR | [29] |
pMLRM1 | icsA (Kpn I - Sal I) gene cloned into pBAD33 (pBAD33::icsA), CmR | This study |
pMLRM2 | icsA ß (amino acid 764-1102) (Not I - Sal I) cloned into pMLRM1 (pBAD33::icsA β ); CmR | This study |
pMAL-c2 | pMAL-c2 (malE); AmpR | New England Biolabs |