I have been trying (unsucessfully) to express a mammalian esterase in
Pichia pastoris both as a fusion to the S. cerevisiae alpha-mating factor
and as an intracellular protein. I have sequenced and everything looks
fine. I have read that some eukaryotic proteins cannot be expressed due
to premature termination at AT-rich regions within the gene of interest.
My question is what is considered to be an "AT-rich" region (i.e., is it a
certain number of consecutive As or Ts, or larger area of high AT content,
or both). I do have a sequence "AAATTTTTAA" which has caught my attention
as a possible problem area.
Any ideas or opinions??
Thanks.
Steve
