Problem about gene leakage in P.pastoris - (Aug/17/2004 )

Hi !
I have expressed allergen protein from dust mite, cysteine protease, in P.pastoris system by using 0.5% induction. At the first time that I set up the experiment, this protein can be expressed with good yield and low in backgroud. After I performed in large scale, nothing can be detected. I think it may be the effects of leakage, so I need the advice how I do.

Thanks

leakage should not happen in this case because your cloned gene is (usually) integrated into the genomic DNA of P. pastoris.