TABLE 3.
Strains and plasmids used in this work
| Strain or plasmid | Genotype/characteristics | Use | Reference(s) or source |
|---|---|---|---|
| Strains | |||
| Escherichia coli | |||
| DH5α | fhuA2 Δ(argF-lacZ)U169 phoA glnV44 ϕ80Δ(lacZ)M15 gyrA96 recA1 relA1 endA1 thi-1 hsdR17 | Cloning | 63 |
| ET12567/pUB307 | dam13::Tn9 dcm6 hsdM hsdR recF143 zjj201::Tn10 galK2 galT22 ara14 lacY1 xyl5 leuB6 thi1 tonA31 rpsL136 hisG4 tsx78 mtli glnV44 F−, carries plasmid pUB307 | Intergeneric conjugation | 64, 65 |
| Streptomyces | |||
| CS149 | Wild type | 1 | |
| ΔsipS15 mutant | ΔsipS15∷aac(3)-IV | This work | |
| ΔsipS4 mutant | ΔsipS4∷aac(3)-IV | This work | |
| ΔsipS9 mutant | ΔsipS9∷aac(3)-IV | This work | |
| ΔsipS14 mutant | ΔsipS14∷aac(3)-IV | This work | |
| ΔsipS8 mutant | ΔsipS8∷aac(3)-IV | This work | |
| ΔsipS5 mutant | ΔsipS5∷aac(3)-IV | This work | |
| ΔsipS13 mutant | ΔsipS13∷aac(3)-IV | This work | |
| ΔsipO1 mutant | ΔsipO1∷hyg | This work | |
| ΔsipO2 mutant | ΔsipO2∷hyg | This work | |
| 149esipR1 | ermE*p-sipR1 | This work | |
| Plasmids | |||
| pSETec | Integrating vector, aac(3)-IV oriTRK2 intΦC31 attPΦC31 ermE*p | Gene overexpression | 66 |
| pSETeTc | Integrating vector, aac(3)-IV tsr oriTRK2 intΦC31 attPΦC31 ermE*p | Gene overexpression | 66 |
| pLHyg | lacZ bla hyg | Source of hyg gene | 57 |
| pUK21 | lacZ km | Backbone for pUKHyg construction | 58 |
| pUKHyg | lacZ km hyg | Replacement cassette construction | This work |
| pUO9090 | aac(3)-IV km | Replacement cassette construction | 67 |
| pHZ1358 | bla tsr oriTRK2 | Knockout mutant generation | 68 |
| pUO149ΔsipS15 | Carries up- and downstream flanking regions of sipS15 at both sides of aac(3)-IV km | Source of sipS15 replacement cassette | This work |
| pUH149ΔsipS15 | Carries sipS15 replacement cassette, aac(3)-IV bla tsr oriTRK2 | sipS15 replacement | This work |
| pSETT149esipS15 | Carries sipS15 under control of ermE*p aac(3)-IV tsr oriTRK2 intΦC31 attPΦC31 | sipS15 complementation | This work |
| pUO149ΔsipS4 | Carries up- and downstream flanking regions of sipS4 at both sides of aac(3)-IV km | Source of sipS4 replacement cassette | This work |
| pUH149ΔsipS4 | Carries sipS4 replacement cassette, aac(3)-IV bla tsr oriTRK2 | sipS4 replacement | This work |
| pSETT149esipS4 | Carries sipS4 under control of ermE*p aac(3)-IV tsr oriTRK2 intΦC31 attPΦC31 | sipS4 complementation | This work |
| pUO149ΔsipS9 | Carries up- and downstream flanking regions of sipS9 at both sides of aac(3)-IV km | Source of sipS9 replacement cassette | This work |
| pUH149ΔsipS9 | Carries sipS9 replacement cassette, aac(3)-IV bla tsr oriTRK2 | sipS9 replacement | This work |
| pSETT149esipS9 | Carries sipS9 under control of ermE*p aac(3)-IV tsr oriTRK2 intΦC31 attPΦC31 | sipS9 complementation | This work |
| pUO149ΔsipS14 | Carries up- and downstream flanking regions of sipS14 at both sides of aac(3)-IV km | Source of sipS14 replacement cassette | This work |
| pUH149ΔsipS14 | Carries sipS14 replacement cassette, aac(3)-IV bla tsr oriTRK2 | sipS14 replacement | This work |
| pSETT149esipS14 | Carries sipS14 under control of ermE*p aac(3)-IV tsr oriTRK2 intΦC31 attPΦC31 | sipS14 complementation | This work |
| pUO149ΔsipS8 | Carries up- and downstream flanking regions of sipS8 at both sides of aac(3)-IV km | Source of sipS8 replacement cassette | This work |
| pUH149ΔsipS8 | Carries sipS8 replacement cassette, aac(3)-IV bla tsr oriTRK2 | sipS8 replacement | This work |
| pSETT149esipS8 | Carries sipS8 under control of ermE*p aac(3)-IV tsr oriTRK2 intΦC31 attPΦC31 | sipS8 complementation | This work |
| pUO149ΔsipS5 | Carries up- and downstream flanking regions of sipS5 at both sides of aac(3)-IV km | Source of sipS5 replacement cassette | This work |
| pUH149ΔsipS5 | Carries sipS5 replacement cassette, aac(3)-IV bla tsr oriTRK2 | sipS5 replacement | This work |
| pSETT149esipS5 | Carries sipS5 under control of ermE*p aac(3)-IV tsr oriTRK2 intΦC31 attPΦC31 | sipS5 complementation | This work |
| pUO149ΔsipS13 | Carries up- and downstream flanking regions of sipS13 at both sides of aac(3)-IV km | Source of sipS13 replacement cassette | This work |
| pUH149ΔsipS13 | Carries sipS13 replacement cassette, aac(3)-IV bla tsr oriTRK2 | sipS13 replacement | This work |
| pSETT149esipS13 | Carries sipS13 under control of ermE*p aac(3)-IV tsr oriTRK2 intΦC31 attPΦC31 | sipS13 complementation | This work |
| pUK149ΔsipO1 | Carries up- and downstream flanking regions of sipO1 at both sides of hyg km | Source of sipO1 replacement cassette | This work |
| pHH149ΔsipO1 | Carries sipO1 replacement cassette, hyg bla tsr oriTRK2 | sipO1 replacement | This work |
| pSET149esipO1 | Carries sipO1 under control of ermE*p aac(3)-IV oriTRK2 intΦC31 attPΦC31 | sipO1 complementation | This work |
| pUK149ΔsipO2 | Carries up- and downstream flanking regions of sipO2 at both sides of hyg km | Source of sipO2 replacement cassette | This work |
| pHH149ΔsipO2 | Carries sipO2 replacement cassette, hyg bla tsr oriTRK2 | sipO2 replacement | This work |
| pSET149esipO2 | Carries sipO2 under control of ermE*p aac(3)-IV oriTRK2 intΦC31 attPΦC31 | sipO2 complementation | This work |
| pSET149eLuxR | Carries sipR1 under control of ermE*p aac(3)-IV oriTRK2 intΦC31 attPΦC31 | sipR1 overexpression | This work |