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Bacterial Infections

Identification of a Chlamydia trachomatis Serovar E Urogenital Isolate Which Lacks the Cryptic Plasmid

Diane R. Stothard, James A. Williams, Barbara Van Der Pol, Robert B. Jones
Diane R. Stothard
Department of Medicine, Indiana University School of Medicine, Indianapolis, Indiana 46202
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James A. Williams
Department of Medicine, Indiana University School of Medicine, Indianapolis, Indiana 46202
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Barbara Van Der Pol
Department of Medicine, Indiana University School of Medicine, Indianapolis, Indiana 46202
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Robert B. Jones
Department of Medicine, Indiana University School of Medicine, Indianapolis, Indiana 46202
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DOI: 10.1128/IAI.66.12.6010-6013.1998
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    Fig. 1.

    omp1 and pCT PCR of strains C599 and E/UW5. Primers designed for omp1 are located 108 bp upstream of the ATG initiator and ∼80 bp downstream of the TAA terminator, resulting in a PCR product of approximately 1,250 bp. Primers designed for pCT are located in ORF8, producing a PCR product of approximately 720 bp. Lanes: 1, 1-kb DNA size standards (Life Technologies, Gibco BRL); 2,omp1 PCR of C599; 3, omp1 PCR of E/UW5; 4, pCT PCR of C599; 5, pCT PCR of E/UW5; 6, negative omp1 control PCR (no template); 7, negative pCT control PCR (no template). Black-and-white digital images were captured with an Alpha Innotech Alpha-Imager 2000 and annotated by using Adobe Photoshop and Freehand V.8 for Power Macintosh.

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    Fig. 2.

    Southern hybridization of BamHI-digested C599 and E/UW5 total DNA probed with DIG-labeled omp1 PCR product (A) and DIG-labeled pCT (B). Lanes: 1, DIG II marker bands at (from top to bottom) 23, 9, 6, 4, 2.3, and 2 kb; 2, 1 μg of E/UW5BamHI-digested DNA; 3, 1 μg of C599BamHI-digested DNA; 4, 0.5 μg of E/UW5BamHI-digested DNA; 5, 0.5 μg of C599BamHI-digested DNA; 6, BamHI-linearized pCT (7.5 kb); 7, omp1 PCR product (1,250 bp). Southern blot X-ray images were scanned at 400 dots per inch with a UMax Powerbook scanner. The images were then combined into one photo by using Adobe Photoshop and Freehand V.8 for Power Macintosh.

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    Fig. 3.

    Fluorescent micrograph of chlamydial inclusion bodies stained with a fluorescein-labeled serovar E-specific anti-major outer membrane protein monoclonal antibody. McCoy cell monolayers were infected with laboratory strain E/UW5 (A) and clinical isolate C599 (B). Infected monolayers were then fixed and stained as previously described (8). Magnification, ×100.

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  • Table 1.

    Growth rates at two independent times for clinical isolate C599 and laboratory strain E/UW5

    TrialNo. of IFU/100 μl
    E/UW5C599
    Primary passageSecondary passagePrimary passageSecondary passage
    123786114625
    22181,7571191
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Identification of a Chlamydia trachomatis Serovar E Urogenital Isolate Which Lacks the Cryptic Plasmid
Diane R. Stothard, James A. Williams, Barbara Van Der Pol, Robert B. Jones
Infection and Immunity Dec 1998, 66 (12) 6010-6013; DOI: 10.1128/IAI.66.12.6010-6013.1998

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Identification of a Chlamydia trachomatis Serovar E Urogenital Isolate Which Lacks the Cryptic Plasmid
Diane R. Stothard, James A. Williams, Barbara Van Der Pol, Robert B. Jones
Infection and Immunity Dec 1998, 66 (12) 6010-6013; DOI: 10.1128/IAI.66.12.6010-6013.1998
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KEYWORDS

Chlamydia Infections
Chlamydia trachomatis
Male Urogenital Diseases
plasmids

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