title: Expression and Purification of Recombinant CDKs: CDK7, CDK8, and CDK9.
abstract: Cyclin-dependent kinases have established roles in the regulation of cell cycle, in gene expression and in cell differentiation. Many of these kinases have been considered as drug targets and numerous efforts have been made to develop specific and potent inhibitors against them. The first step in all of these attempts and in many other biochemical analyses is the production of highly purified and reliable kinase, most frequently in a recombinant form. In this chapter we describe our experience in the cloning, expression, and purification of CDKs using CDK7/CycH, CDK8/CycC, and CDK9/CycT1 as an example.
authors.affiliation: Department of Molecular Biology and Genetics, University of Guelph, 50 Stone Road East, Guelph, ON, Canada, N1G 2W1.
authors.forename: Reena
authors.initials: R
authors.lastname: Pinhero
authors.forename: Krassimir
authors.initials: K
authors.lastname: Yankulov
mesh_terms: D000818:Animals; D002384:Catalysis; D002454:Cell Differentiation; D002845:Chromatography; D056850:Cyclin-Dependent Kinase 8 / Q000737:chemistry / Q000302:isolation & purification*; D042863:Cyclin-Dependent Kinase 9 / Q000737:chemistry / Q000302:isolation & purification*; D018844:Cyclin-Dependent Kinases / Q000737:chemistry / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism; D005786:Gene Expression Regulation; D005982:Glutathione Transferase / Q000378:metabolism; D007313:Insecta; D008967:Molecular Biology / Q000379:methods; D009532:Nickel / Q000737:chemistry; D010455:Peptides / Q000737:chemistry; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification*
chemical_list: D010455:Peptides; D011994:Recombinant Proteins; D009532:Nickel; D005982:Glutathione Transferase; D056850:Cyclin-Dependent Kinase 8; D042863:Cyclin-Dependent Kinase 9; D018844:Cyclin-Dependent Kinases