Problems:
Cathal in his blog post is favoring a linear transfer, but most of the literature is strongly against transformation with linear DNA. So, now I am confused in choosing between a plasmid transfer with chemical/ natural competence or a linear transfer?
-The main difference here is going to be permanence. Plasmids must be selected for to be maintained. Typically, researchers return to a frozen stock of bacteria that they know contains their plasmid each time they want to use it. Your purpose requires many generations of bacteria, possibly over days or weeks, giving ample time for the bacteria to recombine or "spit out" the parts of the plasmid that aren't benefiting it, aka luciferase. The linear transfer is often referred to as "recombineering" when using recombinogenic viral proteins such as lambda Red E/T to enhance linear transfer effeciency, not sure if similar viruses have been studied in acidophilus. Check out Stewart or Murphy for the best recombineering literature.
I am searching for a constitutive promoter for the purpose and would really appreciate any help with this.
-Seems the bacteria will be in lactose, so the lac promoter may be preferred.
Is there anything which I am missing or might have not looked over. Your suggestions please! :)
-dont forget positive / negative selections during cloning, they will make or break this procedure. I'm also not sure how much H202 will be floating around in the yogurt itself if that's where you're hoping the bioluminescence will happen.
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