2008•Yixue fenzi shengwuxue zazhiRequires access
Expression and Purification of Human GST-PTEN Fusion Protein
Xiaohua Liu
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Abstract
Objective To construct a prokaryotic expression vector efficiently expressing PTEN and purify PTEN fusion protein.Methods The PTEN gene was inserted into the prokaryotic GST fusion protein expression plasmid pGEX-4T-1,to form pGEX-4T-1-PTEN.The recombinant plasmid was transformed into E.coli BL2l.After the PTEN protein was induced with IPTG,its expression was analyzed by SDS-PAGE and western blotting.GST-PTEN fusion protein was purified with Glutathion-Sepharose 4B beads.Results The prokaryotic cell expression vector pGEX-4T-1-PTEN was successfully constructed.Highly expressed and purified GST-PTEN fusion protein was obtained.The specificity of fusion protein was verified by SDS-PAGE and western blotting.Conclusion The preparation of GST-PTEN fusion protein can be used to study the biological function of PTEN and its interaction with other proteins.