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.
Balyimez et al. BMC Microbiology 2013 13:269 doi:10.1186/1471-2180-13-269