TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Relevant genotype | Source or reference |
|---|---|---|
| E. coli strains | ||
| XL1-Blue | recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 lac[F′ proAB lacIqZΔM15 Tn10(Tetr)] | Stratagene |
| GM2929 | dam dcm recF hsdR | E. coli Genetic Stock Centera; 38 |
| K12 TB1 | F−ara Δ(lac-proAB) [φ80dlac Δ(lacZ)M15] rpsL(Strr) thi hsdR | New England Biolabs |
| HU227 | hemA41 relA1 spoT1 metB1 rrnB-2 mcrB1 creC510 Hem-p | E. coli Genetic Stock Center |
| Streptomyces strains | ||
| S. nodosus subsp. asukaensis ATCC 29757 | Wild-type strain | Heinz Flossb |
| S. xanthochromogenus AM 6201 | Wild-type strain | Heinz Floss |
| S. parvulus Tü64 | Wild-type strain | Heinz Floss |
| S. bambergiensis ATCC 13879 | Wild-type strain | DSMZ |
| S. ghanaensis ATCC 14672 | Wild-type strain | DSMZ |
| S. lividans TK24 | Wild-type strain | David Hopwoodc |
| S. coelicolor J1501 | hisA1 uraA1 strA1 pgl-1 SCP1- SCP2- | David Hopwood |
| S. nodosus MIP11 | gtr::aph1 | This work |
| S. nodosus MIP12 | hemA-asuA::apr | This work |
| S. nodosus MIP13 | hemA-asuA::apr gtr::aph1 | This work |
| S. coelicolor MIP14 | gtr::aph1 | This work |
| E. coli plasmids | ||
| pTZ19R | Cloning vector | Amersham Biosciences |
| pSL1180 | Cloning vector | Amersham Biosciences |
| pGEM-7Zf(+) | Cloning vector | Promega |
| pBluescript II SK(+) | Cloning vector | Stratagene |
| pQE31 | His-tagged expression vector | QIAGEN |
| pMAL-c2 | MBP-tagged expression vector | New England Biolabs |
| pNEO2 | aph1 from pIJ61 in pBluescript II SK(+) | This work |
| pAPR3 | aac(3)IV from pKC505 in pBluescript II SK(+) | This work |
| pIJ4070 | ermE* promoter in pTZ19R | David Hopwood |
| E68 | Cosmid clone with the gtr locus, S. coelicolor genomic library | 42 |
| pGF23 | Murine hemA gene-carrying plasmid | Gloria C. Ferreira; 14 |
| PMPH22 | gtr PCR fragment in pTZ19R | This work |
| pMPH25 | hemA-asuA PCR fragment in pTZ19R | This work |
| pMPA03 | hemA-asuA region; PstI fragment in pTZ19R | This work |
| pMPA01 | hemA-asuA region; BamHI fragment in pTZ19R | This work |
| pMPA10 | gtr region, BamHI fragment in pTZ19R | This work |
| PMPA11 | gtr gene; KpnI fragment in pTZ19R | This work |
| PMPH28 | BglII-BamHI fragment of E68 in pGEM-7Zf(+) | This work |
| pALS3 | hemA-asuA in pQE31 | This work |
| pALS5 | hemA-asuA in pMAL-c2 | This work |
| pGM160 | E. coli-Streptomyces shuttle; temperature-sensitive replicon; Tsrr | Günter Muth, University of Tübingen, Germany |
| Streptomyces plasmids | ||
| pIJ622 | pIJ702 derivative with an EcoRI linker inserted between mel and tsr genes | David Hopwood |
| pIJ61 | Streptomycete low-copy-number vector | David Hopwood; 17 |
| pIJ487 | Streptomycete high-copy-number vector | 54 |
| pALS4 | ermE*-(6xHis)hemA-asuA fusion in pIJ622 | This work |
| pALS1 | hemA-asuA in pIJ487 | This work |
| pALS2 | hemA-asuA ORFA in pIJ487 | This work |
| PMPA12 | gtr region; KpnI fragment in pIJ487 | This work |
a
New Haven, Conn.
b
University of Washington, Seattle, Wash.
c
John Innes Centre, Norwich, United Kindgom.