TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Description | Reference/source |
|---|---|---|
| E. coli strains | ||
| CAW400 | MC4100 valR recA56 | 35, 46 |
| CC191 | MC1000 F′128(lacIqZΔM15) phoAΔ20 rpoB argE(Am) recA′ | 37, 38 |
| CC1254 | MC1000 F42 ΔrecBCD::red-kan phoAΔ20 rpoB argE(Am) | C. Manoil |
| HS3169 | MC4100 malKΔ16 zjb-729::Tn10 | 33 |
| XK1502 | F− ΔlacU169 nalA | 34 |
| AG1 | F−endA1 hsdR17 (rK− mK+) supE44 thi-1 recA1 gyrA96 relA1 | Stratagene |
| Plasmids | ||
| F′42 | F′lac JCFL0 | 1 |
| F′ΔI | F′42 with traI deleted and replaced by Tn10 tetRA sequence | This study |
| p99D+ | pTrc99A with wild-type traD cloned into the MCSa | This study |
| p99M+ | pTrc99A with wild-type traM cloned into the MCS | This study |
| p99I+ | pTrc99A with wild-type traI cloned into the MCS | This study |
| pACYC184 | Cmr cloning vector with p19A rep functions | 5 |
| pACYCM+ | pACYC184 with wild-type traM | This study |
| pEG100 | pACYC184-derived vector with MCS from pTrc99A | This study |
| pEG103 | pEG103 with traD cloned into the MCS | This study |
| pGK111M0 | 1.2-kb BglII-Pst fragment of plasmid R1 containing oriT, traM, and finP in pUC119; traM not expressed due to G-to-C substitution in start codon ATG | 36 |
| pLOW2 | Kmr vector with p15A rep functions and MCS from pUC18 NotI | 12 |
| pLOW2traM0oriT | EcoRI-Pst oriT-traM0 fragment of pGK111M0 in pLOW2 | This study |
| pNLK5 | Apr vector carrying traD under araBAD promoter | 21 |
| pKD46 | Apr plasmid with λ Red genes gam, bet, and exo | 7 |
| pOXΔtraI | Gmr pOX38 derivative with dhfr gene in place of traI | 46 |
| pPD1 | pHUB2-derived plasmid carrying traD and traI | 43 |
| pRFM2T75 | Cloning vector carrying traM under control of a T7 promoter | L. S. Frost |
| pTrc99A | Apr cloning vector | 2 |
a
MCS, multiple cloning site.