title: Expression in Escherichia coli: purification and characterization of cyclin H, a subunit of the human general transcription/DNA repair factor TFIIH.
abstract: The human cyclin H, a protein normally associated with the cyclin-dependent kinase cdk7, was overexpressed in Escherichia coli using a T7 RNA polymerase expression system and further purified to apparent homogeneity. The purified recombinant cyclin H is similar to the endogenous one according to the following criteria: molecular weight, microsequencing and mass spectra studies, ability to interact with cdk7, and regulatory kinase activity. The scale-up of cyclin H purification is described.
authors.affiliation: Institut de Génétique et de Biologie Moléculaire et Cellulaire, CNRS/INSERM/ULP, Illkirch, France.
authors.forename: A
authors.initials: A
authors.lastname: Poterszman
authors.forename: G
authors.initials: G
authors.lastname: Andersen
authors.forename: D
authors.initials: D
authors.lastname: Busso
authors.forename: M
authors.initials: M
authors.lastname: Rossignol
authors.forename: J M
authors.initials: JM
authors.lastname: Egly
authors.forename: J C
authors.initials: JC
authors.lastname: Thierry
mesh_terms: D000345:Affinity Labels; D000595:Amino Acid Sequence; D015153:Blotting, Western; D056748:Cyclin H; D016213:Cyclins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification*; D004926:Escherichia coli / Q000737:chemistry / Q000235:genetics*; D005822:Genetic Vectors / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D011485:Protein Binding / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification; D019473:Transcription Factors, TFII / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification
chemical_list: D000345:Affinity Labels; C534751:CCNH protein, human; D056748:Cyclin H; D016213:Cyclins; D011994:Recombinant Proteins; D019473:Transcription Factors, TFII