Dear All,
I'm focusing on expressing my fusion protein in Pichia pastoris. The
size of my protein is about more than 200 kDa, and there are mbp-tag and
GFP-his-tag at N-terminus and C-terminus. Verify that my pPICZb plasmid is no
problem by sequencing.
After centrifugation, load the supernatant to amylose column and NI column,
only NI beads have green fluorescence. No strips in the eluent of amylose, and
a strip of gfp-tag for NI column in SDS-PAGE.
Only C-terminal GFP-tag of my fusion protien was expressed !
Why pichia pastoris expresses C-terminal GFP-tag, but neglect my protein
sequence? What should I do to solve this problem and get my protein in Pichia
Pastoris?
Would be very grateful for any advice!
Thanks,
Dong Xiao
########################################################################
To unsubscribe from the CCP4BB list, click the following link:
https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1
