Gal1 promoter construct of mine

* at sbsu1 * at sbsu1.auckland.ac.nz
Tue Nov 21 05:43:18 EST 1995


Dear net readers,
Help! I have encountered a problem with my cloning.  I have made a
construct by amplifying an ORF and cloning into a Gal1-expression vector
(pYES2 for those familiar with the Invitrogen vector).  The gene has a
scorable phenotype, which I expected to be Galactose-regulated in strains
with the new vector.  The gene works fine on galactose, but appears to be
still expressed on glucose plates.  Tried other strains with the same
result.  I included maybe 20 bp of the 5' untranslated region in the
construct - could I have some cryptic promoter still in there?  What is
the basal level of expression of the Gal1 promoter on glucose?  I had read
it was very tightly regulated.  Any suggestions/comments would be
appreciated.

cheers, 
Colin MacDiarmid

SBS, New Zealand.



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