title: Purification and characterization of an enhancer-binding protein of the fibroin gene. I. Complete purification of fibroin factor 1.
abstract: An enhancer-binding protein of the fibroin gene, fibroin factor 1 (FF1), has been purified to homogeneity from crude nuclear extracts of posterior silk gland cells where this gene is transcribed specifically. There is a multiplicity of FF1; the FF1 activity was eluted as at least three major fractions on column chromatographies. FF1 is able to form a stable complex with the enhancer DNA sequence in the presence of another proteinous factor named FF2, which lacks ability to bind DNA molecules by itself. One of FF1 forms, FF1a, was purified with a combination of classical purification techniques without using a sequence-specific affinity column, and identified as a protein with molecular mass 125 kDa using sodium dodecyl sulfate-polyacrylamide gel electrophoresis. To obtain homogeneous protein of FF1a, purification of more than 26,000-fold from the starting nuclear extract was necessary.
authors.affiliation: Department of Developmental Biology, National Institute for Basic Biology, Okazaki, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Suzuki
authors.forename: K
authors.initials: K
authors.lastname: Matsuno
authors.forename: S
authors.initials: S
authors.lastname: Takiya
authors.forename: K
authors.initials: K
authors.lastname: Ohno
authors.forename: K
authors.initials: K
authors.lastname: Ueno
authors.forename: Y
authors.initials: Y
authors.lastname: Suzuki
mesh_terms: D000818:Animals; D001483:Base Sequence; D001667:Binding, Competitive; D012831:Bombyx; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D004247:DNA / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000302:isolation & purification / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D004742:Enhancer Elements, Genetic; D005349:Fibroins / Q000235:genetics*; D008969:Molecular Sequence Data
chemical_list: D004268:DNA-Binding Proteins; D004247:DNA; D005349:Fibroins