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  • Clickable methionine as a universal probe for labelling intracellular bacteria.

Clickable methionine as a universal probe for labelling intracellular bacteria.

Journal of microbiological methods (2019-12-22)
Sharanjeet Atwal, Suparat Giengkam, Yanin Jaiyen, Heather A Feaga, Jonathan Dworkin, Jeanne Salje
ABSTRACT

Despite their clinical and biological importance, the cell biology of obligate intracellular bacteria is less well understood than that of many free-living model organisms. One reason for this is that they are mostly genetically intractable. As a consequence, it is not possible to engineer strains expressing fluorescent proteins and therefore fluorescence light microscopy - a key tool in host-pathogen cell biology studies - is difficult. Strain diversity also limits the universality of antibody-based immunofluorescence approaches. Here, we have developed a universal labelling protocol for intracellular bacteria based on a clickable methionine analog. Whilst we have applied this to obligate intracellular bacteria, we expect it to be useful for labelling free living bacteria as well as other intracellular pathogens.

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Minimum Essential Medium Eagle, With Earle′s salts, non-essential amino acids, L-glutamine and sodium bicarbonate, liquid, sterile-filtered, suitable for cell culture