title: High-level synthesis in Escherichia coli of shortened and full-length human acidic fibroblast growth factor and purification in a form stable in aqueous solutions.
abstract: A highly efficient expression for human acidic fibroblast growth factor (aFGF) has been assembled to direct the synthesis of both shortened and native full-length aFGF. The full-length aFGF-154 form of the protein had not been produced before in Escherichia coli by genetic engineering, and is obtained with its initiator methionine removed. The high production of the aFGF allows one to circumvent the use of reversed-phase chromatography (RPC) during the purification procedure. Here, it is shown that RPC, routinely used to obtain pure preparations of recombinant aFGF, modifies its chemical and physical properties in an unfavorable manner.
authors.affiliation: Centro de Investigaciones Biológicas, C.S.I.C., Madrid, Spain.
authors.forename: M
authors.initials: M
authors.lastname: Zazo
authors.forename: R M
authors.initials: RM
authors.lastname: Lozano
authors.forename: S
authors.initials: S
authors.lastname: Ortega
authors.forename: J
authors.initials: J
authors.lastname: Varela
authors.forename: R
authors.initials: R
authors.lastname: Díaz-Orejas
authors.forename: J M
authors.initials: JM
authors.lastname: Ramírez
authors.forename: G
authors.initials: G
authors.lastname: Giménez-Gallego
mesh_terms: D016475:3T3 Cells; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid; D002942:Circular Dichroism; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics*; D016220:Fibroblast Growth Factor 1 / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D005798:Genes, Bacterial; D005822:Genetic Vectors; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010957:Plasmids; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D012996:Solutions; D013050:Spectrometry, Fluorescence
chemical_list: D000596:Amino Acids; D011994:Recombinant Proteins; D012996:Solutions; D016220:Fibroblast Growth Factor 1