Figure 1

PA2782 and PA2783 constitute an operon. (A) Diagram of the two genes showing their relative size, spacing, and direction of transcription (left to right). Location of the primer pairs, 2782F1-2782R1 and 2782F1-2783R2 (black arrows), and the sizes of the expected products are indicated on the diagram. (B) PCR products obtained from RT-PCR experiments. Overnight culture of PAO1 was subcultured into fresh LB to a starting OD₆₀₀ of 0.02 and incubated to OD₆₀₀ 0.37. Total RNA was extracted from the cells, purified, and used in reverse transcription reactions to produce cDNA. The cDNA was used as a template in PCR reactions with the primer pairs indicated in (A). PAO1 genomic DNA was extracted and used as a positive control and RNA without reverse transcription was used as a negative control. PCR products were separated on 0.8% agarose and stained with ethidium bromide. Lanes: 1) 100-bp molecular size standard, 2) cDNA plus primers 2782F1-2782R1, 3) RNA without reverse transcriptase plus primers 2782F1-2782R2, 4) genomic DNA plus primers 2782F1-2782R2, 5) cDNA plus primers 2782F1-2783R2.