Embarssing problem

[Maurizio Affer]

Hi there
I'm trying to express a small peptide in E. coli.
After doing my cloning in Dh5alpha using a pgex-6p1
vector (amersham) driven by a TAc prmoter I moved the
plasmid into a Bl21-De3 strain.
The problem is I discovered that I'm not even able to
induce expression of GST (which is in the vector of
course)!
I induce at an OD of 0.8 with IPTG final of 0.1mM for
2 hours.
All the trouble shooting are about "not enough protein
induced", but here I'm at "no GST induction"!
Please help whatever comes to your mind let me know...


Maurizio Affer PhD
Memorial Sloan-Kettering Institute
Zuckerman Building
Room 1731B
New York (NY)
10065
Tel.646-888-2072
afferm from mskcc.org



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