Table 1 Strains, plasmids and primers used

Rhodococcus

 R. opacus PD630

Wildtype strain

[39]

 44B2

tadD::Tn5

This study

 DPM245

WT + pDPM70

[17]

 DPM278

WT + pDPM91

This study

 DPM279

WT + pDPM92

This study

E. coli

 E. coli EC100D

F ^- mcrA Δ(mrr-hsdRMS-mcrBC) φ80dlacZΔM15 ΔlacX74 araD139 Δ(ara, leu) pir-116(DHFR)

Epicentre

 DPM54

E. coli EC100D + pMQ70

This study

 DPM55

E. coli EC100D + pDPM21

This study

S. cerevisiae INVSc1

MATa his3D1 leu2 trp1-289 ura3-52

Invitrogen

Plasmids

 pMQ70

oriMB1, oriT, bla, araC, ParaBAD, CEN6, URA3

[44]

 pDPM21

pMQ70 + tadD

This study

 pDPM70

oriNG2, CEN6, aacC1, URA3, Psmyc-tetRO

[17]

 pDPM91

pDPM70 + ro07177

This study

 pDPM92

pDPM70 + tadD

This study

Primers

Sequence

 pDPM21for

5′-GCTTGCATGCCTGCAGGTCGACTCTAGAG GATCCCCGGGTACCTTAATGATGATGATGAT GATGTCGTTTGACCCGCAGCACTCTGTAG-3′

 pDPM21rev

5′-CCGTTTTTTTGGGCTAGCGAATTCAGGAG GCTCTCTCTATGAGTATCGCCGCAGATTCTCT GTCC-3′

 pDPM91for

5′CGATCCGCTCGAGGCATGCAGAAAGGAGG CCATATGGGACTGCATGACTGTCCGGGTAGG CGTAAACGGTTTCGGCCG-3′

 pDPM91rev

5′-GCTATGACCATGATTACGCCAAGCTTGGT A CCGAGCTCGGATCAGAGAGACTTGGCGAC GAGACCGATGAGGTCG-3′

 pDPM92for

5′- CGATCCGCTCGAGGCATGCAGAAAGGAG GCCATATGGGACTGCATGAGTATCGCCGCAG ATTCTCTGTCC-3′

 pDPM92rev

5′- CAGCTATGACCATGATTACGCCAAGCTTGGTA CCGAGCTCGGATCATCGTTTGACCCGCAGCACTC TG-3′