Production and characterization of human basic fibroblast growth factor from Escherichia coli.
The cDNA sequence coding for human basic fibroblast growth factor (bFGF) has been cloned downstream of a transcription promoter recognized by the bacteriophage T7 RNA polymerase. Initiation of translation at the fgf gene has been coupled to upstream translation of a fragment of the T7 phi 10 gene. Expression of the fgf gene in this system can lead to an accumulation of approximately 40 mg/liter/A600 unit of bFGF. This material can be purified close to homogeneity from a soluble protein extract on a heparin-Sepharose column. bFGF so obtained has been shown to have bioactivity indistinguishable from human placental fibroblast growth factor in mitogenicity, synthesis of plasminogen activator, and angiogenesis assays.
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics*; D005260:Female; D005346:Fibroblast Growth Factors / Q000235:genetics* / Q000302:isolation & purification; D005796:Genes; D006801:Humans; D008969:Molecular Sequence Data; D010920:Placenta / Q000378:metabolism; D010957:Plasmids; D011247:Pregnancy; D011401:Promoter Regions, Genetic; D014176:Protein Biosynthesis; D011994:Recombinant Proteins / Q000302:isolation & purification; D014158:Transcription, Genetic
D011994:Recombinant Proteins; D005346:Fibroblast Growth Factors; D004247:DNA