Production of bioactive bovine fibroblast growth factor 4 inE. colibased on the common nucleotide sequence of its structural gene in three breeds

Autor: Kano Kasuga, Tomokazu Fukuda, Masayuki Kobayashi, Yuki Mori, Toshihiko Ito, Miki Sakatani, Mari Yokoo, Ken ichi Yamanaka, Ikuo Kojima, Sho Sato, Saiko Sugawara, Masashi Takahashi
Rok vydání: 2013
Předmět:
Zdroj: Animal Science Journal. 84:275-280
ISSN: 1344-3941
DOI: 10.1111/asj.12036
Popis: 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.
Databáze: OpenAIRE
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