Table 1.
Bacterial strains, plasmids, and oligonucleotides used in this study
| Strains and plasmids | Relevant characteristics | Reference |
|---|---|---|
| Strains | ||
| Streptomyces sp. K30 | Wild type producing clear zones on NR latex overlay agar plates | Rose et al. 2005 |
| Streptomyces sp. K30_ lcp ΩKm | lcp knock out mutant, clear zone negative | This study |
| Streptomyces lividans TK23 | Clear zone negative; host strain for heterologous expression | Hopwood 1983 |
| Streptomyces lividans TK23 pIJ6021:: lcp | Producing clear zones on natural latex overlay agar plates; host strain for heterologous expression harboring wild-type lcp from Streptomyces sp. strain K30 | This study |
| Streptomyces lividans TK24 | Clear zone negative; host strain for heterologous expression | Hopwood 1986 |
| Streptomyces lividans TK24 pIJ6021:: lcp | Producing clear zones on natural latex overlay agar plates; host strain for heterologous expression harboring wild-type lcp from Streptomyces sp. strain K30 | This study |
| Saccharopolyspora erythraea | Wild type, clear zone negative; host strain for heterologous expression | DSMZ 40517 |
| Saccharopolyspora erythraea pIJ6021:: lcp | Producing clear zones on natural latex overlay agar plates; host strain for heterologous expression harboring wild-type lcp from Streptomyces sp. strain K30 | This study |
| Pseudomonas putida | Wild type, clear zone negative; host strain for heterologous expression | DSMZ 291 |
| Pseudomonas putida KT2440 | Clear zone negative; host strain for heterologous expression; pWW0-, r-, m+; spontaneous mutant from P. putida mt-2 | Bagdasarian et al. 1982 |
| Pseudomonas putida KT2440 pBBR1MCS2::Lcp_His6 | Producing clear zones on natural latex overlay agar plates; host strain for heterologous expression harboring wild-type His - tagged lcp protein from Streptomyces sp. strain K30 | This study |
| Pseudomonas putida KT2440 pJB653::Lcp_His6 | Producing clear zones on natural latex overlay agar plates; host strain for heterologous expression harboring wild type His - tagged lcp protein from Streptomyces sp. strain K30 | This study |
| Pseudomonas putida KT2440StrR | Clear zone negative; host strain for heterologous expression; pWW0-, r-, m+; spontaneous streptomycin resistant mutant from P. putida mt-2 | Bagdasarian et al. 1981 |
| Pseudomonas putida KT2440StrR pBBR1MCS2::Lcp_His6 | Producing clear zones on natural latex overlay agar plates; host strain for heterologous expression harboring wild-type His - tagged lcp protein from Streptomyces sp. strain K30 | This study |
| Pseudomonas putida KT2440StrR pJB653::Lcp_His6 | Producing clear zones on natural latex overlay agar plates; host strain for heterologous expression harboring wild-type His - tagged lcp protein from Streptomyces sp. strain K30 | This study |
| Escherichia coli Top10 | Donor strain | Stratagene |
| Escherichia coli ET12567 | Nonmethylating plasmid donor strain | Flett and MacNeil 1992 |
| Plasmids | ||
| pET23a:: lcp _1 | pET23a harboring the wild-type lcp from Streptomyces sp. strain K30 | This study |
| pBBR1MCS2 | Broad host-range promoter-probe vector, pBBR1MCS2 | Kovach et al. 1995 |
| pBBR1MCS2::Lcp_His6 | Shuttle vector harboring the wild-type His-tagged lcp protein from Streptomyces sp. strain K30 | This study |
| pJB653 | Broad host-range promoter-probe vector, pJB653 | Blatny et al. 1997 |
| pJB653::Lcp_His6 | Shuttle vector harboring the wild-type His-tagged lcp protein from Streptomyces sp. strain K30 | This study |
| pGEM-T Easy | E. coli TA cloning vector; Apr | Promega |
| pIJ6021 | High-copy-number plasmid expression vector; contains a thiostrepton-inducible promoter, PtipA, from Streptomyces lividans 66 | Takano et al. 1995 |
| pIJ6021:: lcp | pIJ6021 harboring wild-type lcp from Streptomyces sp. strain K30 | This study |
| pIJ702 | Plasmid contains the tyrosinase gene and thiostrepton resistance (tsr) gene | Rose et al. 2005 |
| pIJ702:: lcp _1 | pIJ702 harboring wild-type gene and the native promoter region of lcp isolated from Streptomyces sp. strain K30 | This study |
| pIJ702:: lcp | pIJ702 harboring the wild-type lcp from Streptomyces sp. strain K30 | Rose et al. 2005 |
| Oligonucleotides | ||
| PSPNter | CCGAGATCTCGGCAGGACGAACTCCCCG | Rose et al. 2005 |
| PSPCter | CCGAGATCTGGTGCGTCGAGG | Rose et al. 2005 |
| Hya_FW_XbaI | AATCTAGAAATAATTTTGTTTAACTTTAAGAAGGAGATATACATATGAACAACGAAGAAACCTTTTATCAGGCC | This study |
| Hya_RW_NcoI | AACCATGGGCGCCCACGCAATTTTCGGC | This study |
| pqspBBR-for:_Sal | ATATGTCGACCTAAAATGGAGTCATGAACAACGAAGAAAACCTTTTATCAGGCCATG | This study |
| pqspBBR-rev:_Sac | ATATGAGCTCCACCACCATCACCACCATGCTCGGACGGTTCACATCCGGAATATCAATCG | This study |
| pqspJB-for:_Sbf | ATATCCTGCAGGTAAGGAGTCATGAACAACGAAGAAACCTTTTATCAGGCCATG | This study |
| pqspJB-rev:_Sac | TATAGAGCTCCACCACCATCACCACCATGCTCGGACGGTTCACATCCGGAATATCAATCG | This study |
| Lcp_EcoRI_6021 | AAAGAATTCTCAGGACGGGCGGTTGACGTCCGGGGATG | This study |
| Lcp_NdeI_6021 | AAAAAAACATATGGCGATCCGCCTTCCGCCCGGCGCCCCGCG | This study |
| N_Lcp | GGATCCTTACGTCAGTAGGCGTGGTCCAGGCCGTCGGTCGG | This study |
| C_Lcp | GGATCCCGACCGGGATGACGTGCGGCAGTGGGCCC | This study |