Table 2 Bacterial strains, plasmids and oligonucleotides used in this study

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 , Ap^r

[18]

 pUC9::strep-lcp _Rr (SN5759)

Cloning vector for lcp _Rr , Ap^r

this study

 p4782.1 (SN3513)

Mobilizable broad host range

[37]

Expression vector, Km^r

 p4782.1::strep-lcp _K30 (SN5496)

Coding sequence of strep-lcp _K30 under

[18]

Rhamnose promoter control, Km^r

 p4782.1::strep-lcp _Rr (SN5760)

Coding sequence of strep-lcp _Rr under

this study

Rhamnose promoter control, Km^r

Oligonucleotides

 Lcp_Rr-complete_for

GCAGAATCCACATGTCCT

 Lcp_Rr-complete_rev

CGACAAACCCACAGATGA

 Lcp_Rr-mature-PstI_for

GGGCCTGCAGCGGCCCTGGAGGTGGTCGCC

 Lcp_Rr-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