I have protein around 15kDa which i am unable to get in BL21 i tried many strains, but still no success. i also encounter with no induction though i got induced earlier. So can i get protein in picchia ? is there always a problem of hyperglycosylation in it. How do i go about it.
Suggestions..
Hi Please check following link it explains very well
http://www.jove.com/video/1862/expression-recombinant-proteins-methylotrophic-yeast-pichia
There might be glycosylated expressed protein, but you can purify it and deglycosylate it using any deglycosylation kits available from sigma, NEB etc.
Do you have a positive control (i.e. a gene cloned in the same expression vector that does express well in E. coli under the same conditions used for your protein? Also, are you sure the protein is not expressed at all? Have you tried Western blotting?
https://tools.thermofisher.com/content/sfs/manuals/easyselect_man.pdf
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