Table 3.
Plasmids used in this study
| Name | Features | Purpose | Reference |
|---|---|---|---|
| pNEB193 | pUC19 ori, bla +, lacZ+ | Parent vector | New England Biolabs |
| pMP44 | oriR6K, bla+, Pmcr (M. barkeri) hpt, Pmcr (M. voltae) pac | Gene deletion by homologous recombination (markerless) | [58] |
| pJK026A | oriV, repE, sopABC, cat, ϕC31 attB, Pmcr (M. voltae) pac-hpt, PmcrB uidA | Gene insertion into ϕC31 attP site on the chromosome | [59] |
| pMS86 | pac (opt) | synthesized | This study |
| pMS66 | hpt (opt) | synthesized | This study |
| pNB716 | strep-his MCS his-strep expression cassette | synthesized | This study |
| pNB721 | pUC19 ori, bla +, lacZ+, pac (opt) | Promotorless pac (opt) for conditional essentiality test of promoters in M. acetivorans | This study |
| pNB722 | pUC19 ori, bla +, lacZ+, PmcrB(M. voltae) pac (opt) | pac (opt) vector for homologous recombination repair of mutants, KO by homologous recombination (marked) in M. acetivorans | This study |
| pNB723 | pUC19 ori, bla +, lacZ+, Pmcr (M. voltae) pac- hpt | Gene deletion by homologous recombination (markerless) in M. acetivorans | This study |
| pNB724 | pUC19 ori, bla +, lacZ+, PmcrB(M. voltae) pac (opt) | pac (opt) vector for homologous recombination repair of mutants, gene deletion by homologous recombination (marked) (unique SpeI restriction site) in M. acetivorans | This study |
| pNB727 | pUC19 ori, bla +, lacZ+, ϕC31 attB, PmcrB(M. voltae) pac (opt) | operon insertion into ϕC31 attP site on the chromosome | This study |
| pNB729 | pUC19 ori, bla +, lacZ+, ϕC31 attB, PmcrB(M. voltae) pac (opt), strep-his MCS his-strep expression cassette | Expression of native or tagged protein in M. acetivorans | This study |
| pNB730 | pUC19 ori, bla +, lacZ+, ϕC31 attB, PmcrB(M. voltae) pac (opt), strep-his MCS his-strep expression cassette | Expression of native or tagged protein (unique BamHI restriction site) in M. acetivorans | This study |
| pALD1 | pUC19 ori, bla +, lacZ+, PmcrB(M. voltae) pac- hpt, ΔMA4421 deletion fusion | MA4421 deletion by homologous recombination (markerless) in M. acetivorans | This study |
| pSK1 | pUC19 ori, bla +, lacZ+, ϕC31 attB, PmcrB(M. voltae) pac (opt), uidA | Expresses native UidA in M. acetivorans | This study |
| pSK2 | pUC19 ori, bla +, lacZ+, ϕC31 attB, PmcrB(M. voltae) pac (opt), strep-his- uidA-his-strep | Expresses strep-his-UidA-his-strep protein in M. acetivorans | This study |
| pNB735 | pUC19 ori, bla +, lacZ+, ϕC31 attB, PmcrB(M. voltae) pac (opt), strep-his (thrombin) MCS his-strep expression cassette | Expression of native or tagged protein in M. acetivorans. Amino-terminal strep-his tag is cleavable with thrombin protease. | This study |
| pNB737 | pUC19 ori, bla +, lacZ+, ϕC31 attB, PmcrB(M. voltae) pac (opt), his (thrombin) MCS his-strep expression cassette | Expression of native or tagged protein in M. acetivorans. Amino-terminal his tag is cleavable with thrombin protease. | This study |