アブストラクト（30巻1号：The Bulletin of Kanagawa Dental College）

The Bulletin of Kanagawa Dental College

English

Title　： Production of Recombinant Human Osteopontin Using Baculovirus Expression Systems
Subtitle　： Selective Proceedings of 36th General Meeting of Kanagawa Odontological Society, 2001
Authors　： Miwa Horiguchi, Masato Yamauchi, Sadao Sato, Masahiro Saito*, Akira Tsunoda*, Toshio Teranaka*
Authors（kana）　：
Organization　： Department of Orthodontics, Kanagawa Dental College, *Operative Dentistry and Endodontics, Kanagawa Dental College
Journal　： The Bulletin of Kanagawa Dental College
Volume　： 30
Number　： 1
Page　： 67-69
Year/Month　： 2002 / 3
Article　： Report
Publisher　： Kanagawa Odontological Society
Abstract　： Osteopontin (OPN) is a secreted, sialic acid-rich, glycophosphoprotein that is abundant in noncollagenous matrix of bone. In order to reveal the functional role of OPN in bone, recombinant human OPN (rhOPN) was expressed in spodoptera frugiperda 9 (Sf9) using baculovirus expression systems. The cDNA encoding 314 amino acids of hOPN from a human placenta library was cloned into baculovirus transfer vector pFastBac1. rhOPN with His6 fusion peptide at C-terminal end was secreted with high efficiency into serum-free culture medium, which yielded as much as 2mg/liter of culture. The purification of rhOPN was achieved by anion exchange chromatography, followed by purification of cobalt affinity column. rhOPN was identified as a band of 65KDa on SDS-PAGE and western blotting. We observed that rhOPN produced in this system was adapted by posttranslational modification such as glycosylation and phosphorylation. Furthermore, rhOPN was biologically active, as assessed by promotion of C3H10T1/2 cells adhesion.
Practice　： Dentistry
Keywords　： Osteopontin, Baculovirus, Cell adhesion, Bone remodeling