TABLE 3.
List of plasmids and strains used in this study
| Strain or plasmid | Use | Genotype or description | Reference |
|---|---|---|---|
| Strains | |||
| E. coli TG2 | supE hsdΔ5 thi Δ(lac-proAB)F′ [traD36 proAB+lacIqlacZΔM15] Δ(srl-recA)306::Tn10 Tetr | 16 | |
| E. coli C43 | Derived from BL21(DE3), tolerant for lethal protein expression | 1 | |
| M. smegmatis MC2155 | Mutant of M. smegmatis mc26 with increased efficiency for electroporation of plasmid DNA; Ept− Kans | 19 | |
| M. marinum ATCC BAA-535 | Hgr; human patient isolate; genome sequence available at http://www.sanger.ac.uk/Projects/M_marinum/ | 14 | |
| Plasmids | |||
| pBRmer BS2 | Mercury resistance and transport studies in E. coli | Apr; Hgr; the EcoRI-NruI fragment of pUB3466, containing Tn501, cloned into pBR322 (ΔEcoRI-NruI) | 24 |
| pBRmerT | Apr; NotI-NruI deletion in pBRmer BS2 to remove merD, merE, and urf2 from the end of Tn501 | This work | |
| pBRmerH | merH from M. marinum cloned in pBRmerT by using BstBI-BglII ligation | This work | |
| pBRmerΔT | Apr; NotI-NruI deletion in pBRmerΔT BS2 to remove merD, merE and urf2 from the end of Tn501 | This work | |
| pBRmerC | Apr; NotI-NruI deletion in pBRmerC BS2 to remove merD, merE, and urf2 from the end of Tn501 | This work | |
| pBRmerF | Apr; NotI-NruI deletion in pBRmerF BS2 to remove merD, merE, and urf2 from the end of Tn501 | This work | |
| pBRmerH CxxS | Apr; cysteine (Cxx changed to serine) mutants of merH cloned into pBRmerT by using BstBI-BglII ligation | This work | |
| pHBL 1 to 5 | MerH topology study | merH amplified with MerH_Nco I_F and HBL1 to -5 and cloned into the pYZ-TBL vector by using NcoI/BstEII restriction sites; this changed the second amino acid of MerH from T to A. | This work |
| 8d06 | Mercury resistance in mycobacteria | Cmr; pBACe3.6 derivative with fragment of pMM23 plasmid DNA containing the putative mer operon | 7 |
| pVV16 | Kanr; HygBr; expression vector used for constitutive protein expression from a PHsp60 promoter | 23 | |
| pVV16-MerA | merA gene from 8d06 cloned with NdeI/HindIII in pVV16 to allow expression of MerA from PHsp60 | This work | |
| pVV16-MerH | merH gene from 8d06 cloned with NdeI/HindIII in pVV16 to allow expression of MerA from PHsp60 | This work | |
| pVV16-MerAH | merH and merA gene from 8d06 cloned with NdeI/HindIII in pVV16 to allow expression of MerA from PHsp60 | This work |