Hi,
I have tried to knockout a secondary metabolite gene from a fungal species, but even after I screen more than 200 colonies and perform 3 rounds of purification, I could not manage to obtain a non-heterokaryon mutant, which led me to think if it's a lethal KO mutant. I tried to search in the literature for similar examples for secondary metabolite genes, but could not find one. Are there any examples of similar situations for other SM genes?
Hi,
I actually started working on another secondary metabolite gene afterwards, and knocked it out in the first try. I used a different construct building method (OSCAR instead of GATEWAY), but the transformation method was the same for both of them.Before the transformation, I had confirmed my KO construct via sequencing. I know it is a long shot but, is it possible that somehow this gene is required for in vitro growth of my hemibiotrophic plant pathogen? I currently don't have much time for repeat experiments for this gene. All I can do is to find most rational explanation.
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