TABLE 1.
Bacterial strains and plasmids used in this studya
| Strain or plasmid | Description | Relevant characteristic(s) | Reference or source |
|---|---|---|---|
| Strains | |||
| E. coli | |||
| HB101 | F− Δ(gpt-proA)62 leuB6 supE44 ara-14 galK2 lacY1 Δ(mcrC-mrr)rpsL20 xyl-5 mtl-1 recA13 | Strr; rB− mB− | 40 |
| TOP10 | F−mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74 deoR recA1 araD139 Δ(araA-leu)7697 galU galK rpsL endA1 nupG | Strr; cloning host | Invitrogen |
| XL-1 Blue | recA1endA1 gyrA96 thi-1 hsdR17 supE44 relA1 lac [F′proAB lacIqΔM15 Tn10] | Tetr; cloning host | Stratagene |
| SM2-1 | plsC1metC162::Tn10 thr-1 ara-14 Δ(gal-attλ)hisG4 rpsL136 xyl-5 mtl-1 lacY1 tsx-78 eda-50 rfbD1 thi-1 | plsC(Ts) mutant | 11 |
| SJ22 | plsB26plsX50 panD2 zac-220::Tn10 glpD3 glpR glpKi relA1 spoT1 pit-10 phoA8 ompF627 fluA22 fadL701 | Auxotrophic for G3P on medium E | 39 |
| R. capsulatus | |||
| MT1131 | crtD121 Rifr | Wild type | 28 |
| Y262 | GTA overproducer | 51 | |
| SA4 | Δ(olsA::spe) | Sper | 3 |
| SA11 | plsC3498::gm | Gmr | This study |
| SA12 | plsC3498::gm Δ(olsA::spe) | Gmr Sper | This study |
| SA13 | Δ(plsC316::kan) | Kanr | This study |
| SA14 | plsC3498::gm Δ(plsC316::kan) | Gmr Kanr | This study |
| SA15 | Δ(olsA::spe) Δ(plsC316::kan) harboring intact olsA on the plasmid | Sper Kanr Tetr | This study |
| SA16 | Δ(olsA::spe) Δ(plsC316::kan) harboring intact plsC316 on the plasmid | Sper Kanr Tetr | This study |
| Plasmids | |||
| pRK2013 | tra+ (RK2) | Kanr; conjugative helper | 18 |
| pRK415 | Tetr; broad-host-range vector | 19 | |
| pBSII | pBluescript II (KS+) | Ampr | Stratagene |
| pMA117 | ΩKan | Kanr | 15 |
| pCHB::Gmr | Gmr Tetr | K. Zhang and F. Daldal | |
| pBAD/Myc-His A | Ampr; arabinose-inducible vector | Invitrogen | |
| pMRC | 6-kb chromosomal EcoRI fragment containing olsA in pLAFR1 | Tetr | 28 |
| pSEM11 | Δ(olsA::spe) | Tetr Sper | 3 |
| pDML1 | 657-bp PCR product containing plsC3498 cloned into NcoI-EcoRI sites of pBAD/Myc-HisA | Ampr | This study |
| pDML3 | 768-bp PCR product containing plsC3498 cloned into XbaI-KpnI sites of pBSII | Ampr | This study |
| pDML4 | XbaI-HindIII-ΩGm of pCHB::Gm inserted into unique SmaI site of plsC3498 on pDML3 | Ampr Gmr | This study |
| pSEM17 | 828-bp PCR product containing olsA cloned into NcoI-EcoRI sites of pBAD/Myc-HisA | Ampr | This study |
| pSEM18 | NsiI-cut pSEM17 ligated into PstI-cut pRK415 | Tetr Ampr | This study |
| pSEM21 | 1.9-kb PCR product containing plsC316 cloned into XbaI-KpnI sites of pBSII | Ampr | This study |
| pSEM24 | 1.9-kb XbaI-KpnI fragment of pSEM21 cloned into XbaI-KpnI sites of pRK415 | Tetr | This study |
| pSEM25 | 819-bp PCR product containing plsC316 cloned into NcoI-SfuI sites of pBAD/Myc-HisA | Ampr | This study |
| pSEM26 | NsiI-cut pSEM25 ligated into PstI-cut pRK415 | Tetr Ampr | This study |
| pSEM27 | NsiI-cut pDML1 ligated into PstI-cut pRK415 | Tetr Ampr | This study |
| pSEM31 | 578-bp Tth111I-RrsII fragment of plsC316 on pSEM21 replaced with SalI-Ωkan of pMA117 | Ampr Kanr | This study |
| pSEM32 | 2.2-kb BglI fragment of pSEM31 ligated to HindIII-KpnI sites of pRK415 | Tetr Kanr | This study |
| pSEM35 | 1,870-bp PshAI-KpnI fragment of pDML4 cloned into KpnI sites of pRK415 | Tetr Gmr | This study |
a
Abbreviations of antibiotic resistances are as follows: Amp, ampicillin; Gm, gentamicin; Kan, kanamycin; Rif, rifampin; Spe, spectinomycin; Str, streptomycin; Tet, tetracycline.