TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Genotype or relevant characteristic | Ref. or source |
|---|---|---|
| Strains | ||
| E. coli | ||
| BL21[λDE3] codonPlus | F−ompT hsdSB (rB− mB−) dcm met gal(λDE3)endA Hte[argU, ileY,leuW] TetR | Novagen |
| BL21[λDE3] pLysS | F−ompT hsdSB (rB− mB−) gal dcm met (λDE3) pLysS (CmR) | Novagen |
| DH5α | K-12 ϕ80d lacZΔM15 endA1 hsdR17 (rK− mK−) supE44 thi-1 gyrA96 relA1 Δ(lacZYA-argF) U169 | Invitrogen |
| Top10 | F−mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74 recA1 araD139 Δ(ara-leu) 7697 galU galrpsL (StrR) endA1 nupG | Invitrogen |
| JW3533-1 | Δ(araD-araB)567 Δ(rhaD-rhaB)568 lacZ487::rmB-3 LAM- rph-1 hsdR514 wecH744::kan lacZ4787 (KmR) | Yale CGSC |
| N. gonorrhoeae FA1090 | Serum-resistant, proline-requiring strain isolated from a patient with probable disseminated gonococcal infection | 41 |
| B. cereus 10987 | NRS 248; genome sequenced strain | ATCC |
| Plasmids | ||
| pET28▵+) | IPTG-inducible T7 expression vector; N- and C- terminal His6 tag; KmR | Novagen |
| pET30▵+) | IPTG-inducible T7 expression vector; N- and C- terminal His6 tag; KmR | Novagen |
| pET32 Ek/LIC | IPTG-inducible T7 expression vector, N- and C- terminal His6 tag; LIC direction cloning strategy; KmR | Novagen |
| pGEM-T Easy | T-overhang assisted cloning, lacZ gene for blue/white screening; AmR | Novagen |
| pBAD 18-Cm | Arabinose-inducible araBAD expression vector, no RBS or purification tag in vector MCS; CmR | 42 |
| pBAD His-A | Arabinose-inducible araBAD expression vector, N-terminal His6 tag; AmR | Invitrogen |
| pACPM16g | pET30▵+) derivative containing ape2 from FA1090 with a G inserted at the 4th base pair (frame shift mutation) on a NdeI/HindIII fragment; KmR | This study |
| pACPM17 | pBAD His-A derivative containing ape2 from FA1090 with an N-terminal His6-tag on a XhoI/HindIII fragment; AmR | This study |
| pACPM19 | pBAD 18-CM derivative containing ape2 from FA1090 with a C-terminal His6 tag on a NheI/HindIII fragment; CmR | This study |
| pACPM20 | A pACPM19 derivative encoding Ape2 truncated by its N-terminal 26 amino acid residues; CmR | This study |
| pACPM22 | A pACPM21 derivative encoding Ape2 truncated by its N-terminal 36 amino acid residues; AmR | This study |