JoVE Encyclopedia of Experiments
Microbiology
0 views • 2:45 min • October 30th, 2025
Begin with a culture of Neisseria gonorrhoeae, a human-specific pathogen grown on a metal-supplemented nutrient medium.
During human infection, this bacterium expresses outer membrane receptors that bind to host proteins carrying metal ions.
This enables the pathogen to extract essential metals and survive in the host environment.
Streak single colonies onto a fresh culture plate and incubate to obtain actively growing colonies.
Inoculate a single healthy colony into a metal-depleted medium in a baffled flask. Then, incubate with shaking to promote bacterial growth.
Once the culture has doubled, dilute it with fresh metal-depleted medium to lower the cell density.
As the bacteria grow without access to external metals, their internal metal reserves are gradually depleted.
This metal scarcity triggers the transcription of metal-responsive genes that encode outer membrane receptors exhibiting a high affinity for human metal-binding proteins.
The bacteria are now ready for downstream analysis.
Two days before beginning the experiment, streak gonococci from freezer stocks onto GC medium plates for a no more than 24 hour incubation at 37 degrees Celsius.
14 to 16 hours before the growth experimen
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