FIG 3.

Transcripts from the seven genes assessed for the E. chaffeensis wild type and the ECH_0660 and ECH_0665 mutants. (A) Schematic representation of the seven genes on E. chaffeensis wild-type chromosome, and ECH_0660 and ECH_0665 mutants. (B) Reverse transcription-PCR (RT-PCR) targets identified. The primer pairs and their estimated products are represented with arrows and bars, respectively. (C) RT-PCR data presented for the amplicons generated targeting the 7 genes. “D” refers to a positive control with genomic DNA as the template; + and − refer to the RT-PCR assays performed with or without reverse transcriptase, respectively. Molecular weight markers (MWM) were included when resolving the PCR products to help locating specific amplicons. Expected amplicons for the internal coding regions of all seven genes were detected for the wild type when reverse transcriptase was added but were absent for the mutation insertion regions of ECH_0660 and ECH_0665 mutants (PCRs 2 and 11, respectively). Also, amplicons from overlapping coding regions of genes were detected as shown. For both ECH_0660 and ECH_0665 mutants, the transcription of the aadA gene from the Himar1 transposon (Tn) insertion was also observed (PCR 12).