Construction of GST-HDAC1 fusion protein vector and its expression in E.coli

Objective To construct GST-HDAC1 fusion protein expression vector and induce its expression in Escherichia coli().Methods The coding sequence of HDAC1 was amplified from the plasmid pcDNA3.1-HDAC1 by PCR and inserted into pGEX-5X-1 by EcoR1.The positive recombinants were identified by restriction endonuclease digestion and DNA sequencing.Then they were transformed into BL21,induced by IPTG and identified by SDS PAGE.Results The length of the forward fragment was 950bp identified by enzymes digestion and the length of reverse fragment was 500bp.The sequence was compared with that in GenBank for homology and the improved prokaryotic expression plasmid pGEX-5X-1-HDAC1 was successfully constructed.The desired GST-HDAC1 fusion protein was expressed in by Western blot.Conclusion The prokaryotic expression plasmid of HDAC1 was successfully constructed and the fusion proteins was expressed in,which provides the basis for the further research on purifying HDAC1 protein and the relationship with other protein.