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Unsuccessful gal induction-ATG context

JOSE LUIS CARRASCO JIMENEZ jlcarrasco at clon.ibmcp.upv.es
Wed May 9 03:30:13 EST 2001


  Dear netters,

  I am trying to express a heterologous protein in yeast
from its own ATG under the control of the GAL1 promoter.
However, to date I have been unsuccessful. I would
appreciate any hint on the possible reason for that.
On one hand I am concerned about the provision of a sequence
context around the ATG optimal for translation initiation
in yeast. Are the Kozak's rules valid for yeast? My
ATG context is: gaattcgaaaATGGAG

  On the other hand, substitution of 2% glucose by 2% galatose
is enough for induction? I have tried both on plate and
on liquid culture.

  Many thanks in advance.

  Cheers,

  Jose

----------------------------------------------------
Jose L. Carrasco
Instituto de Biologia Molecular y Celular de Plantas
Universidad Politecnica de Valencia
Avenida Los Naranjos, s/n
46022 Valencia (Spain)
Email: jlcarrasco at ibmcp.upv.es
----------------------------------------------------


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