TABLE 1.
Strains and plasmids
| Strain or plasmid | Description | Source or reference |
|---|---|---|
| Strains | ||
| BL21 Star(DE3) | F−ompT hsdSB(rB− mB−) gal dcm rne131 (DE3) | Invitrogen |
| XL1-Blue | recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 lac [F′ proAB lacIqZΔM15 Tn10 (Tetr)] | Stratagene |
| BW25141 | lacIqrrnBT14 ΔlacZWJ16 ΔphoBR580 hsdR514 ΔaraBADAH33 ΔrhaBADLD78galU95 endABT333uidA(ΔMluI)::pir+recA1 | 22 |
| BLd | BL21 ΔdeoA | This study |
| BLdt | BL21 ΔdeoA Δtdk | This study |
| BLdtu | BL21 ΔdeoA Δtdk Δudp | This study |
| BLdtu1 | BLdtu harboring pETD::Td | This study |
| BLdtu2 | BLdtu harboring pETD::TdNr | This study |
| BLdtu23 | BLdtu2 harboring pACD::DU | This study |
| BLdtu24 | BLdtu2 harboring pACD::DUTm | This study |
| BLdtu24 (dut) | BLdtu2 harboring pACD::DUTmUt | This study |
| BLdtug24 | BLdtu24 Δung | This study |
| Plasmids | ||
| pETDuet | ColE1 replicon, bla | Novagen |
| pACYCDuet | P15A replicon, cat | Novagen |
| pKD3 | Template plasmid, derivative of pANTSγ, FRT-flanked cat | 13 |
| pKD20 | λ Red helper plasmid, derivative of pINT-ts, araC-ParaB and γ β exo DNA fragments | 13 |
| pCP20 | bla and cat, ori(Ts), thermal inducible FRT recombinase | 13 |
| pETD::Td | pETDuet T4 TDΔI expression under tac promoter, bla | This study |
| pETD::TdNr | pETDuet T4 TDΔI, T4 nrdCAB operon expression under tac promoter, bla | This study |
| pACD::DU | pACYCDuet udk-dcd operon expression under tac promoter, cat | This study |
| pACD::DUTm | pACYCDuet udk-dcd operon, PBS2 TMPase expression under tac promoter, cat | This study |
| pACD::DUTmUt | pACYCDuet udk-dcd operon, dut, PBS2 TMPase expression under tac promoter, cat | This study |