Compositions and methods for producing polypeptides with a modified glycosylation pattern in plant cells

Abstract

A method of modifying a glycosylation pattern of a polypeptide-of-interest in a plant or plant cell is provided. The method comprising expressing in a plant or plant cell transformed to express at least one glycosidase in a subcellular compartment, a nucleic acid sequence encoding the polypeptide-of-interest, such that the at least one glycosidase and the polypeptide-of-interest are co-localized to the subcellular compartment of the plant or plant cell, thereby modifying the glycosylation pattern of the polypeptide-of-interest in the plant or plant cell.

What is claimed is: 
     
         1 . A method for producing a mammalian growth factor in a plant, the method comprising introducing into a plant or plant cell:
 a nucleic acid sequence encoding the growth factor translationally fused to a signal peptide for localization in a subcellular compartment;   at least one polypeptide which confers an agricultural effective trait; and   a nucleic acid sequence encoding an affinity moiety translationally fused to a cell wall binding peptide and a signal peptide for localization in the subcellular compartment,   
       wherein the affinity moiety is for binding the growth factor, and wherein the subcellular compartment is the apoplast or the endoplasmic reticulum. 
     
     
         2 . The method of  claim 1 , wherein the agricultural effective trait is improved insect resistance, disease resistance, herbicide resistance, increased yield, increased tolerance to environmental stress or any combination thereof, as compared to that of a wild-type plant of the same species. 
     
     
         3 . The method of  claim 1 , wherein the plant is of the species  N. tabacum.    
     
     
         4 . The method of  claim 1 , wherein the cell wall binding peptide is a Transmembrane Domain (TMD). 
     
     
         5 . The method of  claim 1 , wherein the cell wall binding peptide is a Cellulose Binding Domain (CBD). 
     
     
         6 . The method of  claim 1 , wherein the affinity moiety is Protein A, Protein G or Protein L. 
     
     
         7 . The method of  claim 1 , wherein the binding of the affinity moiety to the human polypeptide of interest is releasable by pH change. 
     
     
         8 . A plant characterized by:
 (i) the plant expressing a nucleic acid sequence encoding a mammalian growth factor translationally fused to a signal peptide for localization in a subcellular compartment;   (ii) the plant expressing a nucleic acid sequence encoding an affinity moiety translationally fused to a cell wall binding peptide and a signal peptide for localization in the subcellular compartment; and   (iii) the plant expressing at least one nucleic acid sequence encoding a polypeptide which confers an agricultural effective trait;   wherein the affinity moiety is for binding the growth factor, and wherein the subcellular compartment is the apoplast or the endoplasmic reticulum.   
     
     
         9 . The plant of  claim 8 , wherein the agricultural effective trait is improved insect resistance, disease resistance, herbicide resistance, increased yield, increased tolerance to environmental stress or any combination thereof, as compared to that of a wild-type plant of the same species. 
     
     
         10 . The plant of  claim 8 , wherein the plant is of the species N. tabacum. 
     
     
         11 . The plant of  claim 8 , wherein the cell wall binding peptide is a Transmembrane Domain (TMD). 
     
     
         12 . The plant of  claim 8 , wherein the cell wall binding peptide is a Cellulose Binding Domain (CBD). 
     
     
         13 . The plant of  claim 8 , wherein the affinity moiety is Protein A, Protein G or Protein L. 
     
     
         14 . The plant of  claim 8 , wherein the binding of the affinity moiety to the human polypeptide of interest is releasable by pH change.