TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Descriptiona | Reference or source |
|---|---|---|
| Strains | ||
| 19061 ATCC | Wild type, phase I variant Ampr | Laboratory stock |
| ΔopnS | ΔopnS::Cmr | This study |
| opnS+ | ΔopnS::Tn7/opnS::Gmr | This study |
| 19061/Tn7 | Wild type::Tn7/Gmr | This study |
| ΔopnS/TTn7 | ΔopnS::Tn7/Gmr | This study |
| S17-λpir | recA thi pro hsdR−M+; RP4-2Tc::Mu Km::Tn7 in the chromosome | Laboratory stock |
| β2155 | λpir host used for mating with X. nematophila; thrB1004 pro thi strA hsdS lacZΔM15 (F9 lacZΔM15 lacIqtraD36 proA1 proB1) ΔdapA::erm (Ermr) pir::RP4 [::kan (Kmr) from SM10] | 9 |
| S17-λpir/pER2 | Host strain carrying suicide vector pER2 | D. Saffarini |
| S17-λpir/pER2-ΔopnS::Cmr | Host strain carrying suicide vector pER2-ΔopnS::Cmr | This study |
| β2155/pUC18mini-Tn7T | Tn7 transposon vector | This study |
| HB101 | E. coli strain carrying helper plasmid pRK2013 | M. McBride |
| DH5a-λpir/pTNS2 | E. coli strain carrying helper plasmid pTNS2 | |
| β2155-Tn7T | β2155 carrying pUC18 miniTn7T transposon vector | This study |
| β2155-Tn7T opnS | β2155 carrying pUC18 miniTn7T opnS plasmid for complementation of ΔopnS | This study |
| Plasmids | ||
| pSTblue-1 | Cloning vector; Ampr Kmr | Novagen |
| pER2 | Suicide vector; Gmr, R6K ori | D. Saffarini |
| pSTblue-1ΔopnS::Cmr | Cloning vector containing opnS::Cmr fragment | This study |
| pER2ΔopnS::Cmr | Plasmid pER carrying opnS::Cmr fragment for construction of ΔopnS strain in X. nematophila | This study |
| pUC18mini-Tn7T | Tn7 vector | 6 |
| puC18mini-Tn7T-opnS | Tn7 vector carrying the opnS gene | This study |
| pTNS2 | Helper plasmid for integration into the att site | 6 |
| pRK2013 | Helper plasmid for mating | M. McBride |
a
Abbreviations; Ampr, ampicillin resistance; Kmr, kanamycin resistance; Cmr, chloramphenicol resistance.