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Red Fluorescent Chlamydia trachomatis Applied to Live Cell Imaging and Screening for Antibacterial Agents
Umeå University, Faculty of Medicine, Department of Clinical Microbiology.
Umeå University, Faculty of Science and Technology, Department of Chemistry.
Umeå University, Faculty of Medicine, Department of Molecular Biology (Faculty of Medicine).
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2018 (English)In: Frontiers in Microbiology, ISSN 1664-302X, E-ISSN 1664-302X, Vol. 9, article id 3151Article in journal (Refereed) Published
Abstract [en]

In this study, we describe the application of a transformed Chlamydia trachomatis strain constitutively expressing the red fluorescent protein mCherry, to allow real-time monitoring of the infection cycle and screening for agents that block replication of C. trachomatis. The red fluorescent C. trachomatis strain was detected autonomously without antibody staining and was equally susceptible to doxycycline as the wild type strain. A high-throughput screening assay was developed using the transformed strain and automated fluorescence microscopy. The assay was used in a pilot screen of a 349 compound library containing natural products from Australian flora and fauna. Compounds with anti-chlamydial activity were tested for dose response and toxicity to host cells and two non-toxic compounds had 50% effective concentration (EC50) values in the low micromolar range. Natural products are valuable sources for drug discovery and the identified Chlamydia growth inhibition may be starting points for future drug development. Live cell imaging was used to visualize growth of the red fluorescent C. trachomatis strain over time. The screening assay reduced workload and reagents compared to an assay requiring immunostaining and could further be used to monitor the development of Chlamydia inclusions and anti-chlamydial effect in real time.

Place, publisher, year, edition, pages
Frontiers Media S.A., 2018. Vol. 9, article id 3151
Keywords [en]
Chlamydia trachomatis, high content screening, Australian natural products, antibacterial, anti-chlamydial, mCherry, fluorescence-based screening
National Category
Microbiology
Identifiers
URN: urn:nbn:se:umu:diva-155099DOI: 10.3389/fmicb.2018.03151ISI: 000453657900001OAI: oai:DiVA.org:umu-155099DiVA, id: diva2:1277279
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Swedish Research CouncilAvailable from: 2019-01-10 Created: 2019-01-10 Last updated: 2019-03-06Bibliographically approved

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Mojica, SergioEriksson, Anna U.Bahnan, WaelElofsson, MikaelGylfe, Åsa

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Mojica, SergioEriksson, Anna U.Bahnan, WaelElofsson, MikaelGylfe, Åsa
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Department of Clinical MicrobiologyDepartment of ChemistryDepartment of Molecular Biology (Faculty of Medicine)Molecular Infection Medicine Sweden (MIMS)Umeå Centre for Microbial Research (UCMR)
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Frontiers in Microbiology
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