Hello, I am looking for a protocol to insert a GFP plasmid into lactobacillus in agar medium, does anyone has experience in this?
Do you know what strain you have? A lot of lactobacilli are naturally competent, but not dairy onwe
Otherwise, your options are going to be doing electroporation
We are trying wild strains from a kimchi batch to see if we can cultivate it and use electroporation to insert a plasmid ring and bring it back to the fermented kimchi. I found this protocol, could this be useful? https://journals.asm.org/doi/10.1128/aem.02006-10#T2
What equipment do you have for electroporation?
I would first start by isolating the bacteria on an agar plate, and when you have isolates, try transforming them, and growing them on selective media. Then, once you get them, re-introduce them into the ferment.
The only problem with using GFP on a plasmid, is that it will lose the plasmid very quickly.