TABLE 1.
Plasmids used in the study
| Plasmid | Description and/or construction | Reference or source |
|---|---|---|
| pSL1180 | Apr cloning vector; pUC replicon | 5 |
| pJK3 | pac cassette source | 28 |
| pJK41 | Apr Pmr cloning vector; R6K replicon | 27 |
| pMP44 | Vector used to construct up- and down-region cassettes to delete genes from the M. acetivorans C2A chromosome using the markerless exchange method | 33 |
| pMP47 | HindIII/KpnI-digested up-mtaA1 PCR product (with primers 5-upmtaA and 3-upmtaA) cloned into HindIII/KpnI-digested pSL1180 | This study |
| pMP48 | KpnI/BstBI-digested dn-mtaA1 PCR product (with primers 5-dnmtaA and 3-dnmtaA) cloned into KpnI/BstBI-digested pSL1180 | This study |
| pMP54 | Replacement of 166-bp HindIII/KpnI fragment of pMP48 with 1,027-bp HindIII/KpnI fragment of pMP47 | This study |
| pMP64 | AflII/NotI-digested up-mtbA PCR product (with primers AflII-upmtbA and NotI-upmtbA) cloned into AflII/NotI-digested pMP44 | This study |
| pMP65 | NotI/KpnI-digested dn-mtbA PCR product (with primers NotI-dnmtbA and KpnI-dnmtbA) cloned into NotI/KpnI-digested pMP64 | This study |
| pMP68 | Replacement of 52-bp NruI/AvrII fragment of pMP44 with 1,874-bp SmaI/NruI ΔmtaA1 fragment of pMP54 | This study |
| pMP70 | SpeI/SstI-digested up-mtaA2 PCR product (with primers SpeI-upA2 and SacI-upA2) and SstI/NotI-digested dn-mtaA2 PCR product (with primers SacI-dnA2 and NotI-dnA2) cloned into SstI/NotI-digested pMP44 | This study |
| pAMG82 | Vector for construction of promoter fusions to the uidA gene of E. coli | 20 |
| pAB46 | AscI-digested 1,000-bp upstream region of mtaA1 PCR product (using primers mtaA1rev and mtaA1for) was treated with T4 kinase and cloned into Ecl136II/AscI-digested pAMG82 | This study |
| pAB47 | AscI-digested 1,000-bp upstream region of mtaA1 PCR product (using primers mtaA1rev2 and mtaA1for) was treated with T4 kinase and cloned into Ecl136II/AscI-digested pAMG82 | This study |
| pAB48 | AscI-digested 1,000-bp upstream region of mtaA2 PCR product (using primers mtaA2rev and mtaA2for) was treated with T4 kinase and cloned into Ecl136II/AscI-digested pAMG82 | This study |
| pAB49 | AscI-digested 1,000-bp upstream region of mtbA PCR product (using primers mtbArev and mtbAfor) was treated with T4 kinase and cloned into Ecl136II/AscI-digested pAMG82 | This study |