title: Control of foreign polypeptide localization in specific layers of protein body type I in rice seed.
abstract: Prolamin-GFP fusion proteins, expressed under the control of native prolamin promoters, were localized in specific layers of PB-Is. Prolamin-GFP fusion proteins were gradually digested from outside by pepsin digestion. In rice seed endosperm, protein body type I (PB-I) has a layered structure consisting of prolamin species and is the resistant to digestive juices in the intestinal tract. We propose the utilization of PB-Is as an oral vaccine carrier to induce mucosal immune response effectively. If vaccine antigens are localized in a specific layer within PB-Is, they could be protected from gastric juice and be delivered intact to the small intestine. We observed the localization of GFP fluorescence in transgenic rice endosperm expressing prolamin-GFP fusion proteins with native prolamin promoters, and we confirmed that the foreign proteins were located in specific layers of PB-Is artificially. Each prolamin-GFP fusion protein was localized in specific layers of PB-Is, such as the outer-most layer, middle layer, and core region. Furthermore, to investigate the resistance of prolamin-GFP fusion proteins against pepsin digestion, we performed in vitro pepsin treatment. Prolamin-GFP fusion proteins were gradually digested from the peripheral region and the contours of PB-Is were made rough by in vitro pepsin treatment. These findings suggested that prolamin-GFP fusion proteins accumulating specific layers of PB-Is were gradually digested and exposed from the outside by pepsin digestion.
authors.affiliation: Laboratory of Genetic Engineering, Graduate School of Life and Environmental Sciences, Kyoto Prefectural University, Shimogamo, Kyoto, 606-8522, Japan.
authors.forename: Ai
authors.initials: A
authors.lastname: Sasou
authors.affiliation: Laboratory of Genetic Engineering, Graduate School of Life and Environmental Sciences, Kyoto Prefectural University, Shimogamo, Kyoto, 606-8522, Japan.
authors.forename: Takanari
authors.initials: T
authors.lastname: Shigemitsu
authors.affiliation: Laboratory of Genetic Engineering, Graduate School of Life and Environmental Sciences, Kyoto Prefectural University, Shimogamo, Kyoto, 606-8522, Japan.
authors.forename: Yuhi
authors.initials: Y
authors.lastname: Saito
authors.affiliation: Laboratory of Genetic Engineering, Graduate School of Life and Environmental Sciences, Kyoto Prefectural University, Shimogamo, Kyoto, 606-8522, Japan.
authors.forename: Manami
authors.initials: M
authors.lastname: Tanaka
authors.affiliation: Laboratory of Genetic Engineering, Graduate School of Life and Environmental Sciences, Kyoto Prefectural University, Shimogamo, Kyoto, 606-8522, Japan.|Biotechnology Research Department, Kyoto Prefectural Agriculture, Forestry, and Fisheries Technology Research Center, Kitainayazuma, Seika-cho, Soraku-gun, Kyoto, 619-0244, Japan.
authors.forename: Shigeto
authors.initials: S
authors.lastname: Morita
authors.affiliation: Laboratory of Genetic Engineering, Graduate School of Life and Environmental Sciences, Kyoto Prefectural University, Shimogamo, Kyoto, 606-8522, Japan. masumura@kpu.ac.jp.|Biotechnology Research Department, Kyoto Prefectural Agriculture, Forestry, and Fisheries Technology Research Center, Kitainayazuma, Seika-cho, Soraku-gun, Kyoto, 619-0244, Japan. masumura@kpu.ac.jp.
authors.forename: Takehiro
authors.initials: T
authors.lastname: Masumura
mesh_terms: D008856:Microscopy, Fluorescence; D012275:Oryza / Q000378:metabolism / Q000502:physiology*; D010455:Peptides / Q000378:metabolism* / Q000502:physiology; D010940:Plant Proteins / Q000378:metabolism / Q000502:physiology; D030821:Plants, Genetically Modified; D011993:Recombinant Fusion Proteins; D012639:Seeds / Q000378:metabolism / Q000502:physiology*
chemical_list: D010455:Peptides; D010940:Plant Proteins; D011993:Recombinant Fusion Proteins
keywords: Green fluorescent protein; Prolamin; Protein body type I; Rice; Seed storage protein