Problems with protein expression levels in S.Pombe.

Deirdre Buckley dab at ucc.ie
Wed Nov 17 14:07:12 EST 1999


Hello,

We are having problems with expression levels of proteins expressed from
the ADH promoter, when co-expressed with a protein under the control of the
nmt1 promoter. In thiamine plus, expression levels are significantly higher
(anything from 10-100 fold) than in thiamine minus. This was seen both when
the inducible expression was from an autonomously replicating vector (pRSP)
or from a stably integrated version (pINT5).
This is not due to one particular protein as it has been seen with 4
completely different proteins expressed from the ADH promoter.

Has anyone seen this before? Is the nmt1 promoter so strong that the high
levels of protein expression result in competition for translational
machinery?? 

Any suggestions would be greatly appreciated.

Thanks,
Deirdre








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