title: High accumulation of bioactive peptide in transgenic rice seeds by expression of introduced multiple genes.
abstract: We have developed a simple binary vector construction system for the simultaneous expression of multiple genes in plants. Up to three independent gene cassettes can be easily integrated into one binary vector using the MultiSite Gateway System. Using this system, we produced transgenic rice plants that accumulated high levels of the hypocholesterolaemic peptide lactostatin (IIAEK) in endosperm. Binary vectors were constructed that could accommodate up to three independent modified glutelin gene cassettes encoding multimer lactostatin in the variable regions. Eight construct permutations were used for rice transformation. We measured the accumulation of lactostatin expressed as a glutelin fusion protein in the mature seeds of 105 independent transgenic rice lines. A general correlation was observed between accumulation level and gene number in the vector constructs, indicating that a higher accumulation of lactostatin was obtained from transgenic rice plants containing the maximum number of gene inserts. These results indicate that this strategy is applicable for the selection of transgenic lines containing large amounts of bioactive peptides in rice seeds.
authors.affiliation: Transgenic Crop Research and Development Center, National Institute of Agrobiological Sciences, Kannondai 3-1-3, Tsukuba, Ibaraki 305-8604, Japan.
authors.forename: Yuhya
authors.initials: Y
authors.lastname: Wakasa
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Yasuda
authors.forename: Fumio
authors.initials: F
authors.lastname: Takaiwa
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D015153:Blotting, Western; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel; D015870:Gene Expression; D017343:Genes, Plant; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012275:Oryza / Q000196:embryology / Q000235:genetics / Q000378:metabolism*; D010455:Peptides / Q000378:metabolism*; D030821:Plants, Genetically Modified / Q000196:embryology / Q000235:genetics / Q000378:metabolism*; D012639:Seeds / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid
chemical_list: D017931:DNA Primers; D009842:Oligopeptides; D010455:Peptides; C516979:lactostatin