TABLE 1.
Strains and plasmids used in this study
| Strain, plasmid, or transposon | Genotype or markers; characteristics and uses | Source or reference |
|---|---|---|
| Strains | ||
| Escherichia coli TOP10 | F−mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74 recA1 araD139 Δ(ara-leu)7967 galU galK rpsL (Strr) endA1 nupG | Invitrogen |
| Thiobacillus denitrificans | ||
| ATCC 25259 | Wild type | ATCCa |
| TL001 | hynL::kan | This work |
| TL002 | hynL::kan hydA::gent | This work |
| Plasmids | ||
| pUC19 | pMB1, Ampr; cloning vector | 24 |
| pPA20 | pMB1, IncP Ampr Kanr; shuttle vector | 1 |
| pANT4 | IncQ, Ampr Kanrmob+ | 15 |
| pTnMod-OCm | pMB1, Camrmob+ Tn5 tnp | 11 |
| pTnMod-OGm | pMB1, Gentrmob+ Tn5 tnp | 11 |
| pTnMod-OKm′ | pMB1, Kanrmob+ Tn5 tnp | 11 |
| pTnMod-SmO | pMB1, Spcmr Strrmob+ Tn5 tnp | 11 |
| pTnMod-OTc | pMB1, Tetrmob+ Tn5 tnp | 11 |
| pRR10 | IncP, Amprmob+lacZα | 19 |
| pTL1 | IncP, Ampr, pRR10 with PKan inserted at MCS | This work |
| pTL2 | IncP, Gentr Amps, pTL1 with bla::gent; T. denitrificans expression vector | This work |
| pTL3 | IncP, Gentr, pTL2 with hynL-hupE inserted next to PKan allowing for expression | This work |
| EZ-Tn5 pMOD-2<MCS> | ColE1, Ampr; transposon construction vector | EPICENTRE Biotechnologies |
| pMOD-2<gent> | ColE1, Ampr Gentr with SacI fragment from pTnMod-OGm containing aacC1 inserted at MCS of pMOD-2 | This work |
| pUC19-hynL | pMB1, Ampr, pUC19 with hynL inserted at MCS | This work |
| pUC19-hynL::kan | pMB1, Ampr Kanr, pUC19 with hynL::kan inserted at MCS | This work |
| pUC19-hydA | pMB1, Ampr, pUC19 with hydA inserted at MCS | This work |
| pUC19-hydA::gent | pMB1, Ampr Gentr, pUC19 with hydA::gent inserted at MCS | This work |
| Transposons | ||
| Tn-kan | Kanr, EZ-Tn5<KAN-2> DNA fragment with kanamycin resistance selection marker located between Mosaic End Tn5 transposase recognition sequences | EPICENTRE Biotechnologies |
| Tn-gent | Gentr, EZ-Tn5 pMOD-2<gent> DNA fragment with gentamicin resistance selection marker located between Mosaic End Tn5 transposase recognition sequences | This work |
a
ATCC, American Type Culture Collection, Manassas, VA.