Table 2.
Bacterial strains and plasmids used in this study
| Bacterial strain or plasmid | Genotype/Description | Source |
|---|---|---|
| 1292 | supE hsdS met gal lacY tonA | 32 |
| JC8031 | 1292 ΔtolRA | 32 |
| BW25113 |
lacIq
rrnBT14 ΔlacZWJ16hsdR514 ΔaraBADAH33 ΔrhaBADLD78 |
64 |
| BW25113 nlpI::Kan | BW25113 nlpI::Kan created via the method of Datsenko and Wanner |
65 |
| DHB4 | MC1000 phoR Δ (phoA) PvuII Δ (malF)3 F′[lacIqZYA pro] |
Laboratory stock |
| DHA | DHB4 dsbA::Kan | 66 |
| JCA | JC8031 dsbA::Kan | This study |
| pBAD18-Cm | araBAD promoter; pBR322 ori Cmr | 67 |
| pBAD18-Kan | araBAD promoter; pBR322 ori Cmr | 67 |
| pBAD24 | araBAD promoter; pBR322 ori Ampr | 67 |
| pClyA-His6 |
E. coli clyA carrying a C-terminal 6× polyhistidine tag cloned in pBAD18-Cm |
This study |
| pGFP | gfp-mut2 gene cloned in pBAD18-Cm | This study |
| pClyA-GFP |
clyA gene fused to 5’ end of gfp-mut2 in pBAD18-Cm |
This study |
| pGFP-ClyA |
clyA gene fused to 3’ end of gfp-mut2 in pBAD18-Cm |
This study |
| pΔss-Bla | Mature region of bla gene cloned in pBAD18-Kan |
|
| pClyA-Bla |
clyA gene fused to 5’ end of bla in pBAD18-Kan |
This study |
| pBla-ClyA |
clyA gene fused to 3’ end of bla in pBAD18-Kan |
This study |
| pKEG01 | Flavobacterium sp. opd gene in pET78UF | 68 |
| pOPH | opd gene cloned in pBAD24 | This study |
| pClyA-OPH |
clyA gene fused to 5’ end of opd in pBAD24 |
This study |
| pOPH-ClyA |
clyA gene fused to 3’ end of opd in pBAD24 |
This study |
| pLacZ | lacZ gene cloned in pBAD24 | This study |
| pClyA-LacZ |
clyA gene fused to 5’ end of lacZ in pBAD24 |
This study |
| pB18D | anti-digoxin scFv fused to the 3’ end of lpp-ompA cloned in pBAD18 |
58 |
| pB24D | anti-digoxin scFv fused to the 3’ end of lpp-ompA cloned in pBAD24 |
This study |
| pscFv.Dig | anti-digoxin scFv cloned in pBAD24 | This study |
| pClyA-scFv.Dig |
clyA gene fused to 5’ end of anti-digoxin scFv in pBAD24 |
This study |
| pscFv.Dig-ClyA |
clyA gene fused to 3’ end of anti-digoxin scFv in pBAD24 |
This study |
| pClyA(Δ293–303)-scFv.Dig | clyA with 10 C-terminal residues removed | This study |
| pClyA(Δ156–303)-scFv.Dig |
clyA with 147 C-terminal residues removed |
This study |
| pClyA(Y288G)-scFv.Dig | clyA with Tyr288 residue mutated to Gly | This study |