title: Production of recombinant thanatin in watery rice seeds that lack an accumulation of storage starch and proteins.
abstract: Molecular farming is a promising method for producing materials of commercial interest. Plants can be expected to be appropriate hosts for recombinant protein production. However, production in genetically modified plants has two major challenges that must be resolved before its practical use: insufficient accumulation of products and difficulty in establishing methods for their purification. We propose a simple procedure for the production of a desired protein using watery rice seeds lacking an accumulation of storage starch and proteins, a phenotype induced by the introduction of an antisense SPK. We produced a transgenic rice plant containing a gene for an antimicrobial peptide, thanatin, together with antisense SPK. Bioassay and proteome analysis indicated that recombinant thanatin accumulated in an active form in these watery rice seeds. These results suggest that our system worked effectively for the production of thanatin. This procedure enabled easy removal of impurities and simplified the purification process compared with production in leaves. Our system may therefore be a useful technique for the production of desired materials, including proteins.
authors.affiliation: Department of Biological Science and Technology, Tokyo University of Science, 6-3-1 Niijuku, Katsushika, Tokyo 125-8585, Japan.
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Imamura
authors.affiliation: Iwate Biotechnology Research Center, 22-174-4 Narita, Kitakami, Iwate 024-0003, Japan.
authors.forename: Ken-Taro
authors.initials: KT
authors.lastname: Sekine
authors.affiliation: Faculty of Agriculture, Iwate University, 3-18-8 Ueda, Morioka, Iwate 020-8550, Japan.
authors.forename: Tetsuro
authors.initials: T
authors.lastname: Yamashita
authors.affiliation: Department of Biological Science and Technology, Tokyo University of Science, 6-3-1 Niijuku, Katsushika, Tokyo 125-8585, Japan.
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Kusano
authors.affiliation: Department of Biological Science and Technology, Tokyo University of Science, 6-3-1 Niijuku, Katsushika, Tokyo 125-8585, Japan. Electronic address: shimadah@rs.noda.tus.ac.jp.
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Shimada
mesh_terms: D023181:Antimicrobial Cationic Peptides / Q000096:biosynthesis* / Q000302:isolation & purification; D058634:Molecular Farming / Q000379:methods; D012275:Oryza / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism; D010940:Plant Proteins; D030821:Plants, Genetically Modified / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D055314:Seed Storage Proteins / Q000235:genetics*; D012639:Seeds / Q000235:genetics; D013213:Starch / Q000172:deficiency*
chemical_list: D023181:Antimicrobial Cationic Peptides; D010940:Plant Proteins; D011994:Recombinant Proteins; D055314:Seed Storage Proteins; C098076:thanatin; D013213:Starch
keywords: Molecular farming; SPK; Simple purification; Thanatin; Watery rice seeds