In article <aspang-2210962013020001 at bark-632.berkeley.edu>,
aspang at mendel.berkeley.edu (Anne Spang) wrote:
> Hi!
>> I would like to use brefeldin A in S. cerevisiae. What conc. should be
> used for which cell density? Any help (protocol?) would be appreciated!!
>> Thanks in advance!!
>> Anne
I remember hearing that brefeldin A doesn't normally get into yeast cells,
so most strains are resistant. People usually use erg6 mutants which have
a defective plasma membrane that allows some brefeldin A to leak in. I
took a quick look at Biological Abstracts and saw the following
references:
Shah and Klausner (1993) Brefeldin A reversibly inhibits secretion in
Saccharomyces cerevisiae. J Biol Chem 268:5345- 8
Graham, Scott and Emr (1993) Brefeldin A reversibly blocks early but not
late protein transport steps in the yeast secretoy pathway. EMBO J 12:869-
77
Vogel et al. (1993) Brefeldin A causes a defect in secretion in
Saccharomyces cerevisiae. J Biol Chem 268:3040- 3
These papers probably mention what concenrations and conditions to use.
Hope this helps!
-Daniel
