title: Expression in Escherichia coli: production and purification of both subunits of the human general transcription factor TFIIE.
abstract: Both subunits of the human class II transcription factor TFIIE (rTFIIE alpha and rTFIIE beta) have been overexpressed in Escherichia coli at 26 degrees C using a T7 RNA polymerase expression system and further purified to apparent homogeneity. As in this system rTFIIE alpha was poorly expressed and copurified with a truncated form, we expressed rTFIIE alpha as a fusion protein. These overexpressed subunits of TFIIE are similar to the endogenous subunits according to the following criteria: molecular weight, microsequencing, and transcription activity.
authors: Laboratoire de Génétique Moléculaire des Eucaryotes du CNRS, U184 de Biologie Moléculaire et de Génie Génétique de l'INSERM, Faculté de Médecine, Strasbourg, France.
authors: C
authors: C
authors: Chalut
authors: C
authors: C
authors: Lang
authors: J M
authors: JM
authors: Egly
mesh_terms: D000345:Affinity Labels; D000595:Amino Acid Sequence; D001483:Base Sequence; D012321:DNA-Directed RNA Polymerases; D004926:Escherichia coli / Q000235:genetics; D006639:Histidine; D006801:Humans; D008969:Molecular Sequence Data; D010455:Peptides; D011487:Protein Conformation; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D014157:Transcription Factors / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D019473:Transcription Factors, TFII; D014764:Viral Proteins
chemical_list: D000345:Affinity Labels; D010455:Peptides; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors; D019473:Transcription Factors, TFII; D014764:Viral Proteins; C066644:transcription factor TFIIE; C033223:polyhistidine; D006639:Histidine; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases