Pichia expression

3LCID at QUCDN.QueensU.CA 3LCID at QUCDN.QueensU.CA
Fri Nov 4 14:39:12 EST 1994


I am about to try expressing my protein using the Invitrogen Pichia Pastoris ex
pression system.  I am using the pPIC9 vector which excretes the protein using
the alpha factor signal sequence.  My questionis : has anyone tried messing wi
th the sequence between the signal cleavage site and the first restriction site
 SnaBI.  The manual advises not changing this sequence but when I asked Invitro
gen they said they hadn't tried changing it.  Is the EAEAY after the cleavage s
ite necessary for efficient cleavage? I would like to eliminate as much of this
 sequence as possible so that little or no extra sequence is tagged onto the en
d of my protein.  i would appreciate any suggestions or info on this vector spe
ciffically or alpha factor cleavage in general.
Thanks
Carl DeLuca



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