title: Expression and Purification of Recombinant Mouse Interleukin-4 and -6 from Transgenic Rice Seeds.
abstract: Transgenic rice seed can be utilized as a bioreactor to produce high-value recombinant proteins. Mouse interleukin 4 (mIL-4) and mIL-6 were specifically expressed as secretory proteins in rice endosperm by ligating the N-terminal glutelin B-1 (GluB-1) signal peptide and the C-terminal KDEL endoplasmic reticulum retention signal under control of the endosperm-specific GluB-1 promoter. In the transgenic rice seed, mIL-4 and mIL-6 accumulated in levels up to 0.43 mg/g grain and 0.16 mg/g grain, respectively. The reducing agents and detergents required for extraction from the transgenic rice seeds differed between the two proteins, indicating differences in their intracellular localization within the endosperm cell. Purified mIL-4 and mIL-6 exhibited high activity and very low endotoxin contamination.
authors.affiliation: Preventec, Inc., 1-2 Owashi, Tsukuba, Ibaraki, 305-8634, Japan. fwara@preventec-inc.com.
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Fujiwara
authors.affiliation: Transgenic Crop Research and Development Center, National Institute of Agrobiological Sciences, Kannondai 3-1-3, Tsukuba, Ibaraki, 305-8604, Japan.
authors.forename: Lijun
authors.initials: L
authors.lastname: Yang
authors.affiliation: Transgenic Crop Research and Development Center, National Institute of Agrobiological Sciences, Kannondai 3-1-3, Tsukuba, Ibaraki, 305-8604, Japan.
authors.forename: Fumio
authors.initials: F
authors.lastname: Takaiwa
authors.affiliation: Preventec, Inc., 1-2 Owashi, Tsukuba, Ibaraki, 305-8634, Japan.
authors.forename: Kenji
authors.initials: K
authors.lastname: Sekikawa
mesh_terms: D000818:Animals; D003902:Detergents; D015847:Interleukin-4 / Q000235:genetics* / Q000302:isolation & purification* / Q000378:metabolism; D015850:Interleukin-6 / Q000235:genetics* / Q000302:isolation & purification* / Q000378:metabolism; D051379:Mice; D012275:Oryza / Q000235:genetics* / Q000254:growth & development; D030821:Plants, Genetically Modified / Q000254:growth & development; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012639:Seeds / Q000235:genetics* / Q000378:metabolism
chemical_list: D003902:Detergents; D015850:Interleukin-6; D011994:Recombinant Proteins; D015847:Interleukin-4
keywords: Cytokine; Endotoxin free; Interleukin-4; Interleukin-6; Transgenic rice