Improved recombinant protein production in Arabidopsis thaliana

Abstract

Production and isolation of recombinant proteins are key steps in modern Molecular Biology. Expression vectors and platforms for various hosts, including both prokaryotic and eukaryotic systems, have been used. In basic plant research, Arabidopsis thaliana is the central model for which a wealth of genetic and genomic resources is available, and enormous knowledge has been accumulated over the past years - especially since elucidation of its genome in 2000. However, until recently an Arabidopsis platform had been lacking for preparative-scale production of homologous recombinant proteins. We recently established an Arabidopsis-based super-expression system, and used it for a structural pilot study of a multi-subunit integral membrane protein complex. This review summarizes the benefits and further potential of the model plant system for protein productions.

Abbreviations: Nb, Nicotiana benthamiana; OT, oligosaccharyltransferase.

Keywords: Arabidopsis; Endoplasmic Reticulum; Golgi apparatus; complex-type N-glycans; plasma membrane; protein N-glycosylation; recombinant proteins; trans-Golgi network.

Figure 1.

Workflow of establishing Arabidopsis cell lines for recombinant protein expression. The Arabidopsis thaliana rdr6-11 host is transformed by Agrobacterium-mediated floral transformation. Seeds are harvested and primary transformants screened under proper selection. Resistant plants will be further tested, e.g. by immunoblotting using a specific antibody raised against the transgene product (typically using a single fully-expanded cotyledon), or by fluorescence if the target protein is fused to a fluorescent protein. Ten to twenty seedlings with high transgene expression are sliced into small pieces and placed on callus induction medium. Induced calli are transferred to fresh medium every week (established cell lines).