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Table 1 Strains, plasmids and primers used in this study

From: Biofilm as a production platform for heterologous production of rhamnolipids by the non-pathogenic strain Pseudomonas putida KT2440

Strain, plasmid or primer

Relevant characteristic or sequence

References

Strains

 E. coli DH5α

Host for plasmid maintenance

Laboratory strain

 P. putida KT2440

Wild type strain; used for heterologous expression of rhlAB operon

[44]

 P. aeruginosa PAO1

Wild type strain; used for amplification of rhlAB operon

[45]

Plasmids

 pBK-miniTn7-gfp2

Gmr; P rnB P1 gfpAGA

Delivery vector for miniTn7-gfp2

[40]

 pRK600

Cmr; oriColE1 RK2-mob + RK2-tra +

Helper plasmid in mating

[46]

 pUX-BF13

Apr; mob + oriR6K

Helper plasmid providing Tn7 transposition function in trans

[47]

 pJBA27

gfpmut3*

[48]

 pME6031

Tcr; derivate of pME6010; cloning vector maintained in Pseudomonas strains without selection pressure

[37]

 pVW10

pME6031::gfpmut3*

This study

 pVW13

pVW10 with promoterless rhlAB

This study

 pVW14

pVW10 with rhlAB and its native promoter

This study

 pVW12-SPL1 to pVW12-SPL120

SPL in front of rhlAB

This study

Primers

 Gfp_Fwd

5′-AGATAGAATTCAGATTCAATTGTGAGCGG-3′

 Gfp_rev

5′-AGATACTGCAGTATCAACAGGAGTCCAAGC-3′

 SPL_rhlAB

5′-AGATAAGATCTACN10TTGACAN16TATAATN6CCCG-ATCGGCTACGCGTGAACA-3′

 rhlAB_rev

5′-AGATAGAATTCATCACAGCAGAATTGGCCC-3′

 SPL_con_rhlAB

5′-AGATAAGATCTATCGGCTACGCGTGAACA-3′

 Native_rhlAB

5′-AGATAAGATCTACCCTGCCAAAAGCCTGA-3′

  1. The restriction sites on the primers are indicated by underscore. N indicate a randomised base representing 25% of A, C, G or T