(This viral gene has an N terminal hydrophobic transmembrane domain). We could successfully clone and generate positive yeast transformants using pPICZ alpha secretory vector. It is made sure that the full length gene is in correct frame with the signal sequence and alternate strategy of cloning the gene with native N-terminus was also attempted.
Unfortunately, the detection of the recombinant protein in SDS PAGE or western blot is not yet achieved with varying conditions of induction(Tem.,methanol con. etc)
I am looking for suggestions to express our gene of interest in P.pastoris system.
Hello Ancy,
May I know whether did you managed to eventually express your protein using Pichia?
Finally, after quite a lot of trial and error steps, one day it started working....
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