title: A rapid one-step method to purify baculovirus-expressed human estrogen receptor to be used in the analysis of the oxytocin promoter.
abstract: We have produced a truncated form of the human estrogen receptor (hER) as a fusion protein with glutathione S-transferase (GST) in Spodoptera frugiperda (Sf) cells using the baculovirus expression vector (BEV) system. The protein is correctly produced and can be purified from crude whole-cell extracts by a single-step, batch-wise affinity-purification procedure. We show that this GST-hER fusion protein binds at its DNA-binding site specifically and in a hormone-inducible manner. Furthermore, we used the purified hER to analyze the complex estrogen response element (ERE) in the promoter of the oxytocin-encoding gene.
authors.affiliation: Department of Cell Biology, Baylor College of Medicine, Houston, TX 77030.
authors.forename: J M
authors.initials: JM
authors.lastname: Beekman
authors.forename: A J
authors.initials: AJ
authors.lastname: Cooney
authors.forename: J F
authors.initials: JF
authors.lastname: Elliston
authors.forename: S Y
authors.initials: SY
authors.lastname: Tsai
authors.forename: M J
authors.initials: MJ
authors.lastname: Tsai
mesh_terms: D016367:Baculoviridae / Q000235:genetics; D001483:Base Sequence; D004247:DNA / Q000378:metabolism*; D005822:Genetic Vectors / Q000737:chemistry / Q000235:genetics*; D005982:Glutathione Transferase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D011960:Receptors, Estrogen / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
chemical_list: D011960:Receptors, Estrogen; D011993:Recombinant Fusion Proteins; D004247:DNA; D005982:Glutathione Transferase