TABLE 1.
Strains and plasmids
| Plasmid or strain | Genotypea | Reference or source |
|---|---|---|
| Plasmids | ||
| pSET152 | E. coli-Streptomyces shuttle vector; lacZα oriT; pUC replicon, ΦC31 attB-int; Aprr | 32 |
| pST30 | Integrative plasmid pSET152 containing the resistance genes vanHatAatXat from Actinoplanes teichomyceticus | 34 |
| pSET152Y | Integrative plasmid pSET152 containing the resistance gene vanYn | This work |
| pRT802 | pRT801 derived, ΦBT1 attB-int; Kanr | 33 |
| pRT802Y | Integrative plasmid pRT802 containing the resistance gene vanYn | This work |
| pIJ86 | oriT RK2, ermEp*, rep pIJ101, Aprr | 21 |
| pIJ86Y | Multicopy plasmid pIJ86 containing the resistance gene vanYn under the constitutive ermE* promoter | 21 |
| Escherichia coli strains | ||
| DH5α | F− ϕ80lacZΔM15 Δ(lacZYA-argF)U169 recA1 endA1 hsdR17(rK− mK+) phoA supE44 thi-1 gyrA96 relA1 λ− | 26 |
| ET12567 | DNA methylation-deficient host, dam dcm hsdM hsdS hsdR cat tet | 27 |
| ET12567/pUZ8002 | Derived from the methylation-defective strain ET12567 carrying plasmid pUZ8002 | 28 |
| Nonomuraea sp. strains | ||
| ATCC 39727 (wild type) | Producer of the glycopeptide A40926; vanYn is located in the A40926 biosynthetic cluster dbv | 20 |
| Strain carrying pSET152 | Wild-type strain with pSET152 inserted in the ΦC31 attB site; Aprr | This work |
| Strain carrying pST30 | Wild-type strain with the integrative pST30 containing the resistance genes vanHatAatXat from A. teichomyceticus | This work |
| Strain carrying pSET152Y | Wild-type strain with pST152Y inserted in the ΦC31 attB site; Aprr | This work |
| Strain carrying pRT802 | Wild-type strain with pRT802 integrated in the ΦBT1 attB site; Kanr | This work |
| Strain carrying pRT802Y | Wild-type strain with pRT802Y integrated in the ΦBT1 attB site; Kanr | This work |
| Strain carrying pIJ86 | Wild-type strain with the multicopy pIJ86; Aprr | This work |
| Strain carrying pIJ86Y | Wild-type strain with pIJ86Y; Aprr | This work |
| ΔvanYn strain | vanYn-null mutant, sensitive to glycopeptides; Aprr | 21 |
| ΔvanYn-pRT802 strain | ΔvanYn mutant with pRT802; Aprr Kanr | This work |
| ΔvanYn-pRT802Y strain | ΔvanYn mutant with pRT802Y to complement the deletion; Aprr Kanr | This work |
a
Apr, apramycin; Kan, kanamycin.