ArticleApplied and environmental microbiology2007
Efficient method of cloning the obligate intracellular bacterium Coxiella burnetii.
Article in Applied and environmental microbiology, 2007. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed, 17 citations in OpenAlex.
- A simple method for enrichment of phase I Coxiella burnetii.Journal of microbiological methods · 2023Article
- Recent advances in genetic systems in obligate intracellular human-pathogenic bacteria.Frontiers in cellular and infection microbiology · 2023Review
- Bringing genetics to heretofore intractable obligate intracellular bacterial pathogens: Chlamydia and beyond.PLoS pathogens · 2022Article
- Comparative virulence of diverse Coxiella burnetii strains.Virulence · 2019Article
- Genetic mechanisms of Coxiella burnetii lipopolysaccharide phase variation.PLoS pathogens · 2018Article
- Right on Q: genetics begin to unravel Coxiella burnetii host cell interactions.Future microbiology · 2016Review
- Inhibition of inflammasome activation by Coxiella burnetii type IV secretion system effector IcaA.Nature communications · 2015Article
- Genome sequence of Coxiella burnetii strain Namibia.Standards in genomic sciences · 2014Article
- Advances in genetic manipulation of obligate intracellular bacterial pathogens.Frontiers in microbiology · 2011Article
- Coxiella burnetii phase I and II variants replicate with similar kinetics in degradative phagolysosome-like compartments of human macrophages.Infection and immunity · 2010Article
- Characterization of a Coxiella burnetii ftsZ mutant generated by Himar1 transposon mutagenesis.Journal of bacteriology · 2009Article
- Mariner-based transposon mutagenesis of Rickettsia prowazekii.Applied and environmental microbiology · 2007Article
Corrections and comments
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
Abstract
Coxiella burnetii is an obligate intracellular bacterium that replicates in a large lysosome-like parasitophorous vacuole (PV). Current methods of cloning C. burnetii are laborious and technically demanding. We have developed an alternative cloning method that involves excision of individual C. burnetii-laden PVs from infected cell monolayers by micromanipulation. To demonstrate the cloning utility and efficiency of this procedure, we coinfected Vero cells with isogenic variants of the Nine Mile strain of C. burnetii. Coinhabited PVs harboring Nine Mile phase II (NMII) and Nine Mile phase I (NMI) or Nine Mile crazy (NMC) were demonstrated by immunofluorescence. PVs were then randomly excised from cells coinfected with NMI and NMC by micromanipulation, and PVs harboring both strains were identified by PCR. Fresh Vero cells were subsequently infected with organisms from coinhabited PVs, and the PV excision and PCR screening process was repeated. Without exception, PVs obtained from second-round excisions contained clonal populations of either NMII or NMC, demonstrating that micromanipulation is an efficient and reproducible procedure for obtaining C. burnetii clones.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.