title: Overproduction and single-step purification of GAL4 fusion proteins from Escherichia coli.
abstract: A DNA fragment encoding the yeast GAL4 transcriptional activator DNA-binding domain (amino acids 1-93) was cloned into an Escherichia coli expression vector such that the overproduced protein is tagged with six histidine residues and a factor Xa protease cleavage site. The vector also contains unique restriction sites at the 3' end of the gene to allow the construction of fusion proteins. These fusion proteins can easily be purified to homogeneity and their activity tested in vitro.
authors: Department of Biochemistry and Molecular Biology, Harvard University, Cambridge, MA 02138.
authors: R J
authors: RJ
authors: Reece
authors: R J
authors: RJ
authors: Rickles
authors: M
authors: M
authors: Ptashne
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004274:DNA, Recombinant; D004268:DNA-Binding Proteins; D004926:Escherichia coli; D005656:Fungal Proteins / Q000235:genetics* / Q000302:isolation & purification; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000235:genetics / Q000302:isolation & purification; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification; D015183:Restriction Mapping; D029701:Saccharomyces cerevisiae Proteins; D012995:Solubility; D014157:Transcription Factors
chemical_list: D004274:DNA, Recombinant; D004268:DNA-Binding Proteins; D005656:Fungal Proteins; C052130:GAL4 protein, S cerevisiae; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors