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Production of bioactive bovine fibroblast growth factor 4 in E . coli based on the common nucleotide sequence of its structural gene in three breeds
Author(s) -
Sugawara Saiko,
Ito Toshihiko,
Sato Sho,
Yokoo Mari,
Mori Yuki,
Kasuga Kano,
Kojima Ikuo,
Fukuda Tomokazu,
Yamanaka Kenichi,
Sakatani Miki,
Takahashi Masashi,
Kobayashi Masayuki
Publication year - 2013
Publication title -
animal science journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.606
H-Index - 38
eISSN - 1740-0929
pISSN - 1344-3941
DOI - 10.1111/asj.12036
Subject(s) - gene , biology , nucleic acid sequence , fgf10 , escherichia coli , recombinant dna , microbiology and biotechnology , structural gene , peptide sequence , genetics , biochemistry , gene expression
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.