Table 1. Bacterial strains and plasmids used in this study.
| Strain/plasmid | Description | Reference | |
|---|---|---|---|
| ST | |||
| JOL401 | ST wild type, challenge strain | Lab stock | |
| JOL912 | Δlon, ΔcpxR, Δasd mutant ofST | [27] | |
| JOL1592 | JOL912 containing pJHL65-optA and expressing optB | This study | |
| JOL1607 | JOL912 containing pJHL65-optB and expressing optA | This study | |
| JOL1659 | JOL912 containing pJHL65-LfliC and expressing LfliC | This study | |
| JOL1744 | JOL912 containing pJHL65-Lhly and expressing Lhly | This study | |
| E. coli | |||
| BL21 (DE3) pLysS | F−, ompT, hsdSB (rB−, mB−), dcm, gal, λ (DE3), pLysS, Cmr | Progma, USA | |
| JOL1593 | BL21 with pET-optA | This study | |
| JOL1586 | BL21 with pET-OptB | This study | |
| JOL1682 | BL21 with pET-LfliC | This study | |
| JOL1742 | BL21 with pET-Lhly | This study | |
| JOL232 | F− λ− ϕ80 Δ (lacZYA-argF) endA1 recA1 hadR17 deoR thi-1 glnV44 gyrA96 relA1 ΔasdA4 | Lab stock | |
| JOL1601 | JOL232 containing pJHL65-optA | This study | |
| JOL1902 | JOL232 containing pJHL65-OptB | This study | |
| JOL1658 | JOL232 containing pJHL65-LfliC | This study | |
| JOL1743 | JOL232 containing pJHL65-Lhly | This study | |
| Plasmids | |||
| pET28a (+) | IPTG-inducible expression vector; Kanamycin resistant | Novagen, USA | |
| pET32a | IPTG-inducible expression vector; Kanamycin resistant | Lab stock | |
| pET-optA | pET32a derivative containing optA | This study | |
| pET-optB | pET28a (+) derivative containing optB | This study | |
| pET-LfliC | pET28a (+) derivative containing LfliC | This study | |
| pET-Lhly | pET28a (+) derivative containing Lhly | This study | |
| pJHL65 | asd+ vector, pBR ori, bla SS-based periplasmic secretion plasmid, 6xHis, high copy number | [16] | |
| pJHL65-optA | pJHL65 harboring optA of LI | This study | |
| pJHL65-optB | pJHL65 harboring optB of LI | This study | |
| pJHL65-LfliC | pJHL65 harboring LfliC of LI | This study | |
| Gene | |||
| optA | F: GTC GAC ATT TTT AAT CCT CAA GAT | ||
| R: CTC GAG TTA GAA TCT ATA AGT AGC A | |||
| optB | F: GTC GAC ATT TTT AAT CCT CAA GAT | ||
| R: CTG CAG TTA GAA TCT ATA AGT AGC A | |||
| LfliC | F: CCGAATTCTCTTTGGTCATTAACAACAACATGA | ||
| R: CCG TCG ACG CCA CTA ATG AGT TGG CTTG | |||
| Lhly | F: GAA TTC GCC AAA CAT AAA GTA CGT GCT GAT | ||
| R: AAGCTTACGTTTTTTCAAGTAAATAAGATATCTTG | |||
ST, Salmonella typhimurium; LI, Lawsonia intracellularis; IPTG, isopropyl β-D-1-thiogalactopyranoside; bla SS, beta-lactamase signal sequence.