Table 1 Strain construction description
StepExperimentDescriptionAchievement
1PCRPA1 promoter was introduced in front of the CFP and YFP genes.PCR products called PA1-CFP and PA1-YFP
2CloningPA1-CFP and PA1-YFP were cloned in pKD4.aplasmids called pKD4-CFP and pKD4-YFP
3PCRpKD4-CFP and pKD4-YFP were used as templates. Primers with 50 bases sequences homologous to the E. coli rhaA gene at their 5′ ends were used.PCR products called rhaA-CFP-Kan and rhaA-YFP-Kan
4TransformationThe plasmid pKD46a was electroporated into the REL4548 recipient cells.REL4548 carrying pKD46
5Transformation and homologous recombinationPCR products rhaA-CFP-Kan and rhaA-YFP-Kan were electroporated in REL4548 carrying pKD46.REL4548 CFP-KanR and REL4548 YFP-KanR
6TransformationThe plasmid pCP20a was electroporated into REL4548 CFP-KanR and REL4548 YFP-KanR.REL4548 CFP-KanR pCP20 and REL4548 YFP-KanR pCP20
7Heat ShockKanR cassette were excised from the REL4548 CFP-KanR and REL4548 YFP-KanR genomes.REL4548/CFP and REL4548/YFP
  • aFrom Datsenko and Wanner (2000).