Expression profile of the nine <i>pmp</i> genes and <i>ompA</i> throughout the development of <i>C. trachomatis.</i>

<p>Reference strain L₂/434 is represented in panel A and E/Bour in panel B. Values represent the mean±SEM based on three independent experiments for time points of 2, 6, 12, 18, 24, 36, and 48 h post infection. See <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0000878#s2" target="_blank">methods</a> for details.</p

Predicted <i>pmpF</i> promoter sequence for reference and clinical strains.

<p>Sequences are for reference strains E/Bour and L₂/434, and clinical strains E/537C-05, E/S-141, E/CS-500-96, and L₂. The predicted transcription promoter for <i>pmpF</i> is located within a 100% conserved region of the <i>pmpG/pmpF</i> IGR, where putative -10 and -35 elements are in blue characters and boxed. Potential A/T spacer region is underlined, and the predicted transcription start site is shown in a larger font below a red asterisk. The putative RBS for <i>pmpF</i> is in orange characters, and the putative RNase E cleavage sites are highlighted in grey. Numbers represent positions relative to the start codon of <i>pmpF</i> (highlighted in yellow). The start codon of <i>pmpG</i> is highlighted in blue.</p

Distribution/Location of the putative RNase E cleavage sites within the <i>pmpFE</i> operon coding sequence.

<p>The sequence is for reference strains E/Bour and L₂/434 and for clinical strains E/537C-05, E/S-141, E/CS-500-96 and L₂. Black vertical lines represent all RNase E cleavage sites conserved among all strains under study; green vertical lines show the ones only conserved among the four “E” strains; orange vertical lines represent those specific solely for both L₂ strains. Numbers represent nucleotide positions relative to the start codon of <i>pmpF.</i></p

Oligonucleotide primers used for kRT-PCR

a<p>Primers designed based on each <i>pmp</i> sequence of reference strains E/Bour and L₂/434 <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0000878#pone.0000878-Gomes1" target="_blank">[12]</a>.</p>b<p>Primers designed only for strain L₂.</p>c<p>Primers designed based on the <i>ompA</i> sequence of reference strains E/Bour and L₂/434 (GenBank Accession No. X52557 and M14738, respectively).</p>d<p>Primers designed based on the <i>16SrRNA</i> sequence of reference strains E/Bour and L₂/434 (GenBank Accession No. D85722 and U68443, respectively).</p

Clinical and microbiologic characteristics of female adolescents from whom sera was used for determining the immunoreactivity against rPmpD and rPmpF

a<p>Patients were adolescents 14–19 years of age who had a <i>C. trachomatis</i> infection with only one <i>ompA</i> genotype as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0000878#s2" target="_blank">methods</a> or were uninfected;</p>b<p>The diagnosis of cervicitis was based on physical findings consistent with cervicitis as determined by the examining physician; all adolescents infected with <i>C. trachomatis</i> had cervicitis, and none of these patients complained of any symptoms;</p>c<p>A cervical discharge was noted by the examining physician; none of these patients had clinical signs or symptoms consistent with upper genital tract disease.</p

Dot-Blot of serum immunoreactivity against recombinant (r)PmpD and rPmpF.

<p>Sera was obtained from adolescents singly infected and uninfected with a different <i>C. trachomatis</i> clinical strain as described previously <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0000878#pone.0000878-Gomes3" target="_blank">[17]</a> (see also <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0000878#s2" target="_blank">methods</a>). rPmpD and rPmpF concentrations were standardized for use on the blots. Immunoreactivity to each fusion protein for sera from patients infected with strain Ba (n = 3), D (n = 3), E (n = 8), F (n = 5), G (n = 1), Ia (n = 1), J (n = 2) or K (n = 3) are shown. Of note is that immunoreactivity was consistent for sera from patients infected with the same clinical strain except for strain F (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0000878#pone-0000878-t002" target="_blank">Table 2</a>); all eight patients infected with strain E were reactive to rPmpD.</p

Expression profile of <i>pmp</i> and <i>ompA</i> genes throughout the development of <i>C. trachomatis</i> clinical strains.

<p>(A) Strain L₂ shares the same <i>ompA</i> genotype as L₂/434; and strains E/537C-05 (B), E/S-141 (C) and E/CS-500-96 (D) share the same <i>ompA</i> genotype as E/Bour. Values represent the mean±SEM based on three independent experiments for time points of 2, 6, 12, 18, 24, 36, and 48 h post infection. See <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0000878#s2" target="_blank">methods</a> for details.</p