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. 2011 Jul;79(7):2638–2645. doi: 10.1128/IAI.01132-10

Fig. 2.

Fig. 2.

Reverse transcriptase PCR (RT-PCR) analysis of slyA-EF_3001 transcription. The primers used are listed in Table 2. PCRs were performed on chromosomal DNA (lanes 3, 5, 7, 9, and 11) or cDNA (lanes 2, 4, 6, 8, and 10) as a template. To demonstrate the absence of contaminating DNA in the cDNA preparation, sequences upstream of slyA (EF3002U primers, 1.011 bp) (lanes 2 and 3) and downstream of EF_3001 (EF3001int5′ and EF3002D3′ primers, 728 bp) (lanes 10 and 11), which would not be predicted to be cotranscribed, were amplified. The presence of slyA and EF_3001 transcripts was shown by amplifications of internal fragments of slyA (EF3002int primers, 249 bp) (lanes 4 and 5) and of EF_3001 (EF3001int primers, 100 bp) (lanes 6 and 7). To determine whether transcription can proceed from slyA into EF_3001, amplifications of part of slyA-EF_3001 including the 15-bp intergenic region (EF3002int5′ and EF3001int3′ primers, 778 bp) (lanes 8 and 9) were performed. Lanes 1 and 12 correspond to the 10-kb and 100-bp DNA ladders, respectively.