IN VIVO DE-GLYCOSYLATION OF RECOMBINANT PROTEINS BY CO-EXPRESSION WITH PNGASE F

摘要

Materials and methods for in vivo de-glycosylation of recombinant N-glycosylated proteins by co-expression with bacterial PNGase F (Peptide: N-glycosidase F) in plants, using a transient expression system are described. Methods are described which, for example, produce recombinant proteins of interest in plants in a non-glycosylated form. A method of expressing active bacterial PNGase F in plants also is provided.

主权项

1. A method for generating a deglycosylated polypeptide of interest, comprising producing a eukaryotic cell that comprises:
(a) a first nucleic acid comprising a first nucleotide sequence encoding a bacterial PNGase F polypeptide, wherein the first nucleotide sequence is operably linked to a promoter such that when the promoter is activated, the PNGase F polypeptide is expressed; and (b) a second nucleic acid comprising a nucleotide sequence encoding the polypeptide of interest, wherein the second nucleotide sequence is operably linked to a promoter such that when the promoter is activated, the polypeptide of interest is expressed, wherein by action of the PNGase F polypeptide, the polypeptide of interest is deglycosylated.