Sat Sep 23 17:56:24 EST 1995
In order to map the binding site(s) on a receptor for a peptide hormone I'm
studying, I want to make a library of semi-random point-mutants, express this
library of mutants in yeast, and then screen by FACS (using a fluorescent
derivative of the hormone as a probe) for mutants that don't bind to the
hormone. Our lab does not have mammalian tissue culture facilities (which is
why we want to do our studies in yeast), but there is a departmental FACS.
How permeable is the yeast cell wall? I realize that for immunofluoresence,
the cell wall must be removed. But the peptides of interest are only 30-50
residues long (an order of magnitude smaller than antibodies). Would these have
a problem getting through the cell wall to the plasma membrane (where the
receptor would be localized)?
navin at uclink4.berkeley.edu
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