Table 1 Bacterial strains and plasmids used in this study
Strain/plasmid | Description | Reference |
|---|---|---|
S. Typhimurium | ||
JOL990 | S. Typhimurium wild type, challenge strain | Lab stock |
JOL1800 | Salmonella Typhimurium ∆lon, ∆cpxR, ∆asd, ∆rfaL; bacterial delivery vector; rough strain | Lab stock |
JOL1809 | JOL1800 containing pJHL65 and expressing OptA | This study |
JOL1810 | JOL1800 containing pJHL80 and expressing OptB | This study |
JOL1811 | JOL1800 containing pJHL65 and expressing FliC | This study |
JOL1812 | JOL1800 containing pJHL65 and expressing Hly | This study |
E. coli | ||
BL21(DE3)pLysS | F−, ompT, hsdSB (r −B , m −B ), dcm, gal, λ (DE3), pLysS, Cmr | Promega |
JOL232 | F− λ− ϕ80 ∆(lacZYA-argF) endA1 recA1 hadR17 deoR thi-1 glnV44 gyrA96 relA1 ∆asdA4 | Lab stock |
JOL1601 | JOL232 containing pJHL80-OptA | This study |
JOL1902 | JOL232 containing pJHL65-OptB | This study |
JOL1658 | JOL232 containing pJHL65-FliC | This study |
JOL1743 | JOL232 containing pJHL65-Hly | This study |
Plasmids | ||
pET28a(+) | IPTG-inducible expression vector; Kanamycin resistant | Novagen |
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-fliC | pET28a(+) derivative containing fliC | This study |
pET-hly | pET28a(+) derivative containing hly | This study |
pJHL65 | asd+ vector, pBR ori, β-lactamase signal sequence-based periplasmic secretion plasmid, 6xHis, high copy number | [24] |
pJHL80 | asd+ vector, p15A ori, β-lactamase signal sequence-based periplasmic secretion plasmid, 6xHis, high copy number | |
pJHL-OptA | pJHL80 harboring optA of L. intracellularis | This study |
pJHL-OptB | pJHL65 harboring optB of L. intracellularis | This study |
pJHL-FliC | pJHL65 harboring fliC of L. intracellularis | This study |
pJHL-Hly | pJHL65 harboring hly of L. intracellularis | This study |
Primers | ||
rfaL DEL OT F | 5′-GGATACGATAAACCGCAGTCG | This study |
rfaL DEL OT R | 5′- AACCGTGCGCTTGCTGATAAG | This study |
rfaL DEL IN F | 5′- ACAAGTTTAGGACTTCGCTGCC | This study |
rfaL DEL IN R | 5′-CAGAATGGTATTATGCGGACCG | This study |