TABLE 2.
Plasmids used in this work
| Plasmid | Relevant properties | Reference or source |
|---|---|---|
| E. coli | ||
| pGEM-T Easy | Used for cloning of PCR amplified DNA fragments, Apr | Promega |
| pDR540 | Contains the galK gene under control of the tac promoter, Apr | Accession no. U13847 |
| pSN40 | pRM1 derivative (30) containing an S. lividans TK21 chromosomal BamHI fragment encoding the four SPases, Apr | 32 |
| pEX50 | pDR540 derivative, tac promoter, Apr | This work |
| pET3a | T7 promoter, Apr | Novagen |
| pEX20 | pET3a derivative encoding hexahistidine-tagged SipY336 | This work |
| pEX23 | pET3a derivative encoding hexahistidine-tagged SipY304 | This work |
| pTM | pGEM-T Easy derivative containing the gene encoding mature tendamistat | This work |
| pSipTM | pTM derivative, contains different SPase-TM fusion genes | This work |
| pEXSipY336 | pEX50 derivative encoding wild-type SipY | This work |
| pEXSipY304 | pEX50 derivative encoding SipY derivative lacking the C-terminal anchor | This work |
| Streptomyces | ||
| pAX5a | E. coli-Streptomyces shuttle vector encoding pre-tendamistat, Apr Tsr | 15 |
| pIJ6021 | Contains a multiple cloning site after a tipA promoter, Kmr | 39 |
| pIJSipTM | pIJ6021 derivative encoding the different SPase-TM fusion proteins | This work |