If you want to test the expression of a virus protein in mammalian cells (HEK) and yeasts (P. Pastoris) and you could only optimize the DNA sequence for one of the two cases, which one would you choose?
It is better to test a yeast optimized sequence in mammalian or a mammalian optimized sequence in yeast?
Thanks!
Hi Juan,
My choice would be to optimise for mammalian followed by a sanity check that it doesn't produce any really bad sequences for the Pichia. My reasoning is that a somewhat sub optimal expression level in Pichia is far more easily overcome by increasing the biomass than in mammalian cells.
I think however it is very easy to get too focussed on optimisation of the DNA sequence as a fix to all expression problems. Yes a sequence far from the optimum can badly affect the protein expression, but there are many post translational blockers to the generation of the final protein that can be the limiting factor. So sequence optimisation only helps if it's the rate limiting step.
-Richard
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