GST vector expression of ScFv
lpss at unixg.ubc.ca
Thu Nov 21 07:08:40 EST 1996
I am trying to use a GST vector (Pharmacie, GST-4T2) to express a
single-chain antibody with little success (The orientation of the
vector: GST-Kozak sequence-ScFc).
The inserts were conformed by colony PCR. I have checked to make sure the
fusion protein sequence is in frame with GST. 0.2 mM IPTG used. Induction
was done at 26C for 8 hrs. I've tried two strains of E coli: DH5alpha,
UT5600 (protease deficient).
I'd like to know whether anyone has some clue as to what's going on. Is
Kozak sequence or antibody sequence too toxic for bacteria to express?
University of British Columbia
achang at hivnet.ubc.ca
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