Expression and purification of the human homeodomain oncoprotein HOX11.

摘要: HOX11 is a transcription factor belonging to the homeodomain family that essential for spleen development during embryogenesis. It also tumorigenic, being associated with T-cell acute lymphoblastic leukemia in children. In order understand functional role of both normal and malignancy, protein-DNA protein-protein interaction studies involving this are required. Such investigations would be facilitated by availability significant amounts purified protein. However, expression full-length bacteria has been reported problematic owing fusion protein instability. Here, we report purification human expressed Escherichia coli as soluble glutathione S-transferase (GST) addition, mutant version was produced (HOX11ΔH3) which lacked DNA-recognition helix (helix 3) homeodomain. Through single procedure using glutathione-Sepharose, 2 mg recombinant proteins were obtained per liter bacterial culture. Notably, GST-HOX11 had markedly higher stability when at low temperature (4°C). Purification near-homogeneity achieved judged SDS-PAGE recognized anti-HOX11 antibodies. The biological activity verified specific binding GST-HOX11, but not GST-HOX11ΔH3, DNA containing consensus recognition sites.