TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Relevant characteristic(s) | Source or reference |
|---|---|---|
| Strains | ||
| E. coli | ||
| DH5α | F− φ80dlacZΔM15 Δ(lacZYA-argF)U169 recA1 endA1 hsdR17(rK− mK+) phoA supE44 λ−thi-1 gyrA96 relA1 | Laboratory strain |
| XL1-Blue MR | Δ(mcrA)183 Δ(mcrCB-hsdSMR-mrr)173 endA1 supE44 thi-1 recA1 gyrA96 relA1 lac | Stratagene |
| S. mutabilis subsp. capreolus | Wild type (ATCC 23892) | ATCCa |
| S. lividans 1326 | Wild type | A. Gehring |
| EAF1001 | S. lividans 1326 with pCMN-P4C8RF-436 integrated into the φC31 attB site; clone 1 | This study |
| EAF1002 | S. lividans 1326 with pCMN-P4C8RF-436 integrated into the φC31 attB site; clone 2 | This study |
| EAF1003 | S. lividans 1326 with pOJ436 inserted into the φC31 attB site; clone 1 | This study |
| EAF1004 | S. lividans 1326 with pOJ436 inserted into the φC31 attB site; clone 2 | This study |
| Plasmids | ||
| SuperCos1 | Kanr Ampr cloning cosmid | Stratagene |
| pOJ436 | Aprr oriT φC31 int φC31 attP cosmid vector | 3 |
| pCR-BluntII-TOPO | Kanr cloning vector | Invitrogen |
| pCMN-P4C8RF | S. mutabilis subsp. capreolus genomic DNA cloned into SuperCos1; contains CMN gene cluster | This study |
| pCMN-P4C8RF-436 | pCMN-P4C8RF with Aprr oriT φC31 int φC31 attP DraI fragment of pOJ436 inserted into HpaI site | This study |
| pTOPO-cmnU | cmnU cloned into pCR-BluntII-TOPO cloning vector | This study |
| pBAD33 | Arabinose-inducible expression vector | 14 |
| pET22b | Kanr cloning vector | Novagen |
| pET22b-cmnU | cmnU cloned into NdeI/HindIII site of pET22b | This study |
| pBAD33-cmnU | XbaI/HindIII fragment of pET22b-cmnU cloned into XbaI/HindIII site of pBAD33 | This study |
| pLJ102 | IPTG-inducible expression vector | 18 |
| pSJ101 | pLJ102 derivative with tlyA insert | 18 |
| pSE34 | Streptomyces sp. expression vector | 29 |
| pSE34-cmnU | pSE34 with cmnU cloned into the XbaI/HindIII site | This study |
a
ATCC, American Type Culture Collection.