Expression and Purification of Recombinant CDKs: CDK7, CDK8, and CDK9

Methods Mol Biol. 2016:1336:13-28. doi: 10.1007/978-1-4939-2926-9_3.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Catalysis
  • Cell Differentiation
  • Chromatography
  • Cyclin-Dependent Kinase 8 / chemistry
  • Cyclin-Dependent Kinase 8 / isolation & purification*
  • Cyclin-Dependent Kinase 9 / chemistry
  • Cyclin-Dependent Kinase 9 / isolation & purification*
  • Cyclin-Dependent Kinases / chemistry
  • Cyclin-Dependent Kinases / isolation & purification*
  • Electrophoresis, Polyacrylamide Gel
  • Escherichia coli / metabolism
  • Gene Expression Regulation
  • Glutathione Transferase / metabolism
  • Insecta
  • Molecular Biology / methods
  • Nickel / chemistry
  • Peptides / chemistry
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / isolation & purification*

Substances

  • Peptides
  • Recombinant Proteins
  • Nickel
  • Glutathione Transferase
  • Cyclin-Dependent Kinase 8
  • Cyclin-Dependent Kinase 9
  • Cyclin-Dependent Kinases