Table 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Description | Source or reference |
|---|---|---|
| E. coli strains | ||
| TOP10 | λ− F−mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74 recA1 araD139 Δ(ara-leu)7697 galU galK rpsL (Strr) endA1 nupG | Invitrogen |
| DH5α | λ− φ80dlacZΔM15 Δ(lacZYA-argF)U169recA1 endA1 hsdR17(rK− mK−) supE44 thi-1 gyrA relA1 | 9 |
| BL21(DE3) | F−ompT hsdSB(rB− mB−) gal dcm with DE3, a λ prophage carrying the T7 RNA polymerase gene | Novagen |
| SMhisMrpC2 | BL21(DE3) containing pET16b/His10-MrpC2 | 29 |
| SMFruAhis | BL21(DE3) containing pET11km/FruA-His6 | 29 |
| M. xanthus strains | ||
| DK1622 | Wild type | 13 |
| DK5285 | fruA::Tn5lac Ω4491 | 20 |
| MKV6 | attB::pKV6 (pREG1727 with 150-bp XhoI-BamHI fragment from pKV3)a | 55 |
| MKV24 | attB::pKV24 (pREG1727 with 351-bp XhoI-BamHI fragment from pPV251) | This study |
| MKV54 | attB::pKV54 (pREG1727 with 398-bp XhoI-BamHI fragment from pKV45) | This study |
| MBS03 | attB::pBS08 (pREG1727 with 197-bp XhoI-BamHI fragment from pBS02) | This study |
| MBS04 | attB::pBS07 (pREG1727 with 216-bp XhoI-BamHI fragment from pBS01) | This study |
| MBS06 | attB::pBS21 (pREG1727 with 193-bp XhoI-BamHI fragment from pBS15) | This study |
| MBS07 | attB::pBS18 (pREG1727 with 170-bp XhoI-BamHI fragment from pBS13) | This study |
| MBS08 | attB::pBS19 (pREG1727 with 160-bp XhoI-BamHI fragment from pBS14) | This study |
| Plasmids | ||
| pET16b/His10-MrpC2 | pET16b carrying a gene encoding His10-MrpC2 under the control of a T7 RNA polymerase promoter | 35 |
| pET11km/FruA-His6 | pET11km carrying a gene encoding FruA-His6 under the control of a T7 RNA polymerase promoter | S. Inouye |
| pREG1727 | Apr Kmr; P1-incattP ′lacZ | 7 |
| pCR2.1-TOPO | Apr Kmr; lacZα | Invitrogen |
| pKV3 | pCR2.1-TOPO with fmgE DNA from positions −100 to +50 | 55 |
| pKV4 | pREG1727 with 1.0-kb XhoI-BamHI fragment from pPV4406-1.0 | 55 |
| pKV12 | pCR2.1-TOPO with fmgE DNA from positions −100 to + 320 | 55 |
| pKV28 | pKV12 with CATCGTG-to-ACGATGT mutation from positions −55 to −49 | 55 |
| pKV29 | pKV12 with GGACA-to-TTCAC mutation from positions −69 to −65 | 55 |
| pKV30 | pKV12 with GAACC-to-TCCAA mutation from positions −64 to −60 | 55 |
| pKV45 | pCR2.1-TOPO with fmgE DNA from positions −150 to +251, generated by PCR using LK0926 and LK1245 as primers and pKV4 as template | This study |
| pKV47 | pKV12 with A-to-C mutation at position −54 | 55 |
| pPV251 | pCR2.1-TOPO with fmgE DNA from positions −100 to +251 | 55 |
| pBS01 | pCR2.1-TOPO with fmgE DNA from positions −100 to +116, generated by PCR using LK1328 and LK2527 as primers and pKV45 as template | This study |
| pBS02 | pCR2.1-TOPO with fmgE DNA from positions −150 to +50, generated by PCR using LK0926 and LK1034 as primers and pKV45 as template | This study |
| pBS13 | pCR2.1-TOPO with fmgE DNA from positions −120 to +50, generated by PCR using LK2577 and LK1034 as primers and pKV45 as template | This study |
| pBS14 | pCR2.1-TOPO with fmgE DNA from positions −110 to +50, generated by PCR using LK2578 and LK1034 as primers and pKV45 as template | This study |
| pBS15 | pCR2.1-TOPO with fmgE DNA from positions −100 to +93, generated by PCR using LK1328 and LK2606 as primers and pKV45 as template | This study |
a
Where possible, the plasmid description is given in parentheses after the strain description.