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J Biotechnol ; 108(2): 179-83, 2004 Mar 04.
Article in English | MEDLINE | ID: mdl-15129727

ABSTRACT

The utility of plants for the production of a wide range of recombinant proteins is now clearly established. However, the challenge remains to produce these proteins at sufficient concentrations for extraction to be economically feasible. In this paper, we have investigated the ability of plant cells to accumulate the human interleukin-10 (IL-10) protein targeted to chloroplasts and mitochondria. We found that IL-10 accumulates in chloroplasts only if a 6 x His tag is added at the C-terminus of the protein. The hexapeptide may provide protection from degradation. Conversely, the IL-10 protein does not accumulate in mitochondria. Analysis of the chloroplast-targeted IL-10 protein revealed only monomeric IL-10 and limited biological activity in in vitro cell assays.


Subject(s)
Chloroplasts/genetics , Chloroplasts/metabolism , Gene Targeting/methods , Interleukin-10/biosynthesis , Interleukin-10/genetics , Mitochondria/genetics , Mitochondria/metabolism , Plants, Genetically Modified/metabolism , Humans , Interleukin-10/chemistry , Molecular Weight , Mutagenesis, Site-Directed/genetics , Plant Leaves/genetics , Plant Leaves/metabolism , Protein Engineering/methods , Recombinant Proteins/biosynthesis , Subcellular Fractions/metabolism , Nicotiana/genetics , Nicotiana/metabolism
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