title: Identification of Core Alpha 1,3-Fucosyltransferase Gene From Silkworm: An Insect Popularly Used to Express Mammalian Proteins.
abstract: Silkworm has great potential as production system of recombinant mammalian proteins. When the protein products are used for medical purpose, it is required to reduce the risk of an allergy, the content of core alpha 1,3-fucosyl residue attached to the N-glycan of proteins, for example. We isolated the gene of an enzyme responsible for the transfer of core alpha 1,3-fucosyl residue, core alpha 1,3-fucosyltransferase (Fuc-T C3), from silkworm. A candidate cDNA for silkworm Fuc-T C3 was isolated as a homolog of the fruit fly enzyme gene fucTA. The gene was located on chromosome 7 of the silkworm genome and was composed of seven exons, which spanned approximately 10 kb on the genome. The coding region of the gene was 1,350 bp and encoded a 450-amino acid protein with a molecular mass of 52.2 kDa. Deduced amino acid sequence of the coding region showed one transmembrane domain in its N-terminal and typical motifs common to fucosyltransferases including Fuc-T C3s of other organisms in its C-terminal. The extract of CHO cells transfected with the cDNA showed Fuc-T C3 activity using GDP-fucose and DABS-GnGn peptide as substrates. These results showed this cDNA clone actually encodes silkworm Fuc-T C3.
authors.affiliation: Central Laboratory, Nippon Flour Mills Co., Ltd., 5-1-3 Midorigaoka, Atsugi, Kanagawa, 243-0041, Japan. s-minagawa@nippn.co.jp.
authors.forename: Sachi
authors.initials: S
authors.lastname: Minagawa
authors.affiliation: Central Laboratory, Nippon Flour Mills Co., Ltd., 5-1-3 Midorigaoka, Atsugi, Kanagawa, 243-0041, Japan.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Sekiguchi
authors.affiliation: Central Laboratory, Nippon Flour Mills Co., Ltd., 5-1-3 Midorigaoka, Atsugi, Kanagawa, 243-0041, Japan.
authors.forename: Yuzuru
authors.initials: Y
authors.lastname: Nakaso
authors.affiliation: Transgenic Silkworm Department, Immuno-Biological Laboratories Co., Ltd., 1091-1, Naka, Fujioka, Gunma, 375-0005, Japan.
authors.forename: Masahiro
authors.initials: M
authors.lastname: Tomita
authors.affiliation: Central Laboratory, Nippon Flour Mills Co., Ltd., 5-1-3 Midorigaoka, Atsugi, Kanagawa, 243-0041, Japan.
authors.forename: Manabu
authors.initials: M
authors.lastname: Takahisa
authors.affiliation: Department of Animal Biotechnology, Konkuk University, Seoul 143-701, Korea.
authors.forename: Hideyo
authors.initials: H
authors.lastname: Yasuda
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D012831:Bombyx / Q000201:enzymology / Q000235:genetics*; D016466:CHO Cells; D003412:Cricetulus; D005647:Fucosyltransferases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D049750:Genome, Insect; D019476:Insect Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid
chemical_list: D019476:Insect Proteins; D005647:Fucosyltransferases; C031692:galactoside 3-fucosyltransferase
keywords: Bombyx mori; N-glycan; fucose; glycosylation; transferase