I have been using Zeocin marker for target protein expression to verify integration in Pichia system. However, I would like to remove Zeocin marker since it's antibiotic related and not ideal for food application. Wondering what other marker gene I can use or knockout Zeocin marker from Pichia genome since the plasmid is already integrated.
Thanks for everyone!
I assume you do not want resistance to other antibiotics xD
What strain of Pichia is it? You could use autotrophy/heterotrophy system, but if your strain is autotrophic, you would need to mutate it first (but what selection will you use, right?).
The best option is probably to start over and use some other resistance from the beginning.
Thank you for the answer. I am using Pichia wildtype (Mut+).
You are correct. Ideally we should not use any antibiotics.
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