Transcription level of plasmid-encoded <i>versus</i> chromosomally encoded <i>pomB</i> in <i>V</i>. <i>cholerae</i> determined by qRT-PCR.

<p>A: analysis of amplification products obtained from qRT-PCR reactions with template (cDNA) amounts of 100 ng, 10 ng, 1 ng and 0.1 ng by agarose gel electrophoresis. The efficiency of the primer pair “pomB fwd” / “pomB rev” was 115% with R² = 0.998, as calculated using the CFX Manager software (Bio-Rad). A 100 bp DNA ladder (GeneRuler, Thermo Scientific) served as molecular size marker. B: The correct size (134 bp) of the products from the qRT-PCR reactions (technical triplicates, I–III) was confirmed by agarose gel electrophoresis. C: Relative transcription level of <i>pomB</i> in the <i>V</i>. <i>cholerae</i> reference strain and in <i>V</i>. <i>cholerae</i> Δ<i>pomAB</i> pAB induced with 10 mM L-arabinose. For both strains, qRT-PCR reactions were performed in technical triplicates. Transcription levels of <i>pomB</i> were calculated using the CFX Manager software (Bio-Rad, version 2.1.1022.0523), and the transcription level of the chromosomally encoded <i>pomB</i> (reference strain) was set to 1.