I have bought bacteria transfected with a bioluminescent plasmid (https://www.addgene.org/search/catalog/plasmids/?q=54630).
The plasmid is supposedly constitutively active, and the luciferase should also be automatically active (the plasmid should encode both enzyme and substrate).
Yet, the bacteria do not show any luminescence by the eye, possibly because the light is beyond the eye field. But even under a fluorescent microscope, no light is given off.
How are bioluminescent bacteria activated? How do you see their luminescence?
Thank you
You are aware that the plasmids from that study were designed to study expression plants and not bacteria, so the promoter region is not designed for constitutive expression in bacteria, aren't you? There could be some leaky expression if at all but that would hardly be detectable using a microscope, let alone by the naked eye! For luminescence you need a reader with luminescence detection or at least a gel documentation system with luminescence detection. They are much more sensitive for luminescence detection (there's a reason why you need to expose western blots for a few minutes sometimes to accumulate the weak signal).
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