Production of human basic fgf mutein

Details Production of human basic fgf mutein Production of human basic fgf mutein

C - Chemistry, Metallurgy

12

N

C12N 15/18 (2006.01) C07K 14/50 (2006.01) C12N 1/21 (2006.01) C12N 15/67 (2006.01) C12N 15/70 (2006.01) A61K 38/00 (2006.01)

Patent

CA 2070989

2070989 9109126 PCTABS00005 Disclosed are (1) a vector comprising a nucleotide sequence coding for a mutein in which at least one constituent amino acid of a mature human basic fibroblast growth factor (hbFGF) is replaced by another amino acid, and a T7 promoter upstream therefrom; (2) a transformant transformed by the vector of (1); (3) the transformant of (2), in which a host is E. coli having a T7 RNA polymerase gene downstream from a lac promoter; (4) a method for producing the mutein in which at least one constituent amino acid of the mature hbFGF is replaced by another amino acid, which comprises cultivating the transformant of (2) in a culture medium; (5) the method of (4), in which about 3 to 500 µm of isopropylthiogalactopyranoside is added to the culture medium on a logarithmic growth phase of the transformant of (3), followed by cultivation, and (6) the method of (5), in which a resultant mutein-containing solution is purified by chromatography using a crosslinked polysaccharide sulfate, a synthetic polymer having a sulfonic acid group as an exchange group and/or a synthetic polymer for gel filtration as a carrier, whereby the hbFGF mutein having biological activity can be efficiently produced.

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