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Generation and Multi-phenotypic High-content Screening of Coxiella burnetii Transposon Mutants
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Chapters
- 00:05Title
- 01:49Isolation of Individual Transposon Mutants
- 03:44Evaluation of Bacterial Concentration and Single Primer Colony PCR
- 05:34Eukaryotic Cell Challenge with Coxiella Mutants and Monitoring of Intracellular Growth
- 07:17Preparation of Samples for Automated Image Acquisition
- 09:25Results: Viability and Infectivity of Coxiella Transposon Mutants
- 11:14Conclusion
Coxiella burnetii is an obligate intracellular Gram-negative bacterium responsible for the zoonotic disease Q fever. Here we describe methods for the generation of Coxiella fluorescent transposon mutants as well as the automated identification and analysis of the resulting internalization, replication and cytotoxic phenotypes.
