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JoVE Journal

Immunology and Infection

This content is Open Access.

Generation and Multi-phenotypic High-content Screening of Coxiella burnetii Transposon Mutants

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11:44 min

May 13th, 2015

May 13th, 2015

9,657 Views

0:05

Title

1:49

Isolation of Individual Transposon Mutants

3:44

Evaluation of Bacterial Concentration and Single Primer Colony PCR

5:34

Eukaryotic Cell Challenge with Coxiella Mutants and Monitoring of Intracellular Growth

7:17

Preparation of Samples for Automated Image Acquisition

9:25

Results: Viability and Infectivity of Coxiella Transposon Mutants

11:14

Conclusion

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.

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