Table 2.
Strain or plasmid | Relevant characteristics | Reference |
E. coli JM109 | Plasmid storage and expression of lcp | |
E. coli XL1-blue | Transformation strain | Stratagene |
Rhodococcus rhodochrous RPK1 | Wild type strain, degrades rubber | this study |
pUC9::strep-lcp K30 (SN5339) | Cloning vector for lcp K30, Apr | [18] |
pUC9::strep-lcp Rr (SN5759) | Cloning vector for lcp Rr, Apr | this study |
p4782.1 (SN3513) | Mobilizable broad host range | [37] |
Expression vector, Kmr | ||
p4782.1::strep-lcp K30 (SN5496) | Coding sequence of strep-lcp K30 under | [18] |
Rhamnose promoter control, Kmr | ||
p4782.1::strep-lcp Rr (SN5760) | Coding sequence of strep-lcp Rr under | this study |
Rhamnose promoter control, Kmr | ||
Oligonucleotides | ||
LcpRr-complete_for | GCAGAATCCACATGTCCT | |
LcpRr-complete_rev | CGACAAACCCACAGATGA | |
LcpRr-mature-PstI_for | GGGCCTGCAGCGGCCCTGGAGGTGGTCGCC | |
LcpRr-mature-HindIII_rev | CCGGTAAGCTTTCAGGGATAGTTGGG | |
16S-universal-for | GAGTTTGATC(A/C)TGGCTCAG | |
16S-universal-rev | GG(C/T)TACCTTGTTACGACT | |
16S-Rr-complete_for | CTGGCGCGGTGCTTAAC | |
16S-Rr-complete_rev | CAGTAATTCCGGACAACG |
Kanamycin resistance (Kmr), Ampicillin resistance (Apr)