JoVE Encyclopedia of Experiments
Encyclopedia of Experiments
This video demonstrates a method for assessing the intracellular growth of GFP-tagged Coxiella mutants within cultured epithelial cells. The mutant phenotype is visualized using immunostaining and epifluorescence microscopy, allowing the study of the effect of the mutation on intracellular growth.
1. Generation of a library of green fluorescence protein, GFP-tagged Coxiella transposon mutants
Manipulate Coxiella burnetii RSA439 NMII in a biosafety containment level 2 (BSL-2) in a microbial safety cabinet (MSC) in compliance with local rules. If compatible with the bacterial model used, repeat steps from 1.4.1 to 1.4.4 to increase the probability of obtaining clonal mutants. A typical mutant library is composed (at least) of a number o.......
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Source: Martinez, E. et al., Generation and Multi-phenotypic High-content Screening of Coxiella burnetii Transposon Mutants. J. Vis. Exp. (2015)
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