skuroda at inherit.biosig.kobe-u.ac.jp
Tue Apr 4 18:53:47 EST 1995
> Overexpression studies
> 12:12 PM 4/4/95
> I would like to get some input from the yeast community on overexpressing
> proteins in S. cerevisiae. In general (I know it is strongly protein
> how much protein can be produced by overexpressing a non-yeast gene from
> the Gal1- or ADH-promoter.
It's not a case by either Gal1 or ADH promoter. In 1992, I suceeded in
hepatitis B virus envelope L protein particles in S. cerevisiae by TDH3
of which the level is up to 42% of total soluble protein (see JBC 267 1953
I think this is the highest expression level by S cerevisiae system.
>How did you determine the amount of your expressed
> protein? What is the minimal amount one can detect on Western blots? Does
> anybody know of any references that address this question? I very much
> appreciate all the input and I promise an extensive summary of all the data I
> can collect from you.
First I determine the amount of total proteins by Cu-Folin method.
Second I determine the amount of HBV env L protein particles by densitometric
method by using the purified particles as a standard.
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