title: Production of bioactive bovine fibroblast growth factor 4 in E. coli based on the common nucleotide sequence of its structural gene in three breeds.
abstract: Fibroblast growth factor 4 (FGF4) is considered a crucial gene in the proper development of bovine embryos. We recently determined the FGF4 gene sequence in eight cattle derived from three breeds and revealed a common nucleotide sequence of the structural gene encoding FGF4, which leads to the deletion and mutation of amino acid sequences in the mature FGF4 (Pro(32) -Leu(206) ) compared with the sequence previously reported. In the present study, HisbFGF4, a 6× histidine-tagged bovine FGF4 (Pro(32) -Leu(206) ), was produced in Escherichia coli based on the validated nucleotide sequence and purified by heparin column chromatography. In primary bovine fibroblasts, HisbFGF4 showed significant mitogenic activity, whereas, intriguingly, the activity of a commercially available recombinant human FGF4 (Gly(25) -Leu(206) ) produced in E. coli was weaker than that of HisbFGF4. In conclusion, the present study provides a simple method for the production of a bioactive bovine FGF4 derivative in E. coli utilizing its structural gene elucidated by us.
authors.affiliation: Graduate School of Bioresource Sciences, Akita Prefectural University, Akita, Japan.
authors.forename: Saiko
authors.initials: S
authors.lastname: Sugawara
authors.forename: Toshihiko
authors.initials: T
authors.lastname: Ito
authors.forename: Sho
authors.initials: S
authors.lastname: Sato
authors.forename: Mari
authors.initials: M
authors.lastname: Yokoo
authors.forename: Yuki
authors.initials: Y
authors.lastname: Mori
authors.forename: Kano
authors.initials: K
authors.lastname: Kasuga
authors.forename: Ikuo
authors.initials: I
authors.lastname: Kojima
authors.forename: Tomokazu
authors.initials: T
authors.lastname: Fukuda
authors.forename: Ken-Ichi
authors.initials: K
authors.lastname: Yamanaka
authors.forename: Miki
authors.initials: M
authors.lastname: Sakatani
authors.forename: Masashi
authors.initials: M
authors.lastname: Takahashi
authors.forename: Masayuki
authors.initials: M
authors.lastname: Kobayashi
mesh_terms: D000818:Animals; D001483:Base Sequence; D002417:Cattle / Q000196:embryology*; D049109:Cell Proliferation; D002845:Chromatography; D004926:Escherichia coli / Q000737:chemistry*; D051519:Fibroblast Growth Factor 4 / Q000032:analysis / Q000235:genetics*; D005347:Fibroblasts / Q000166:cytology; D005822:Genetic Vectors; D011506:Proteins / Q000032:analysis
chemical_list: D051519:Fibroblast Growth Factor 4; D011506:Proteins