Expression,Purification and Identification of GST-Dot4 Fusion Protein

[Objective] The paper aimed to improve the soluble expression of Dot4p in E.coli and to obtain higher purity protein.[Method]The pGEX-KG-Dot4 recombinant plasmid was transformed to E.coli BL2l and the expression of GST-Dot4 fusion protein was induced with IPTG.The expression product was identified by SDS-PAGE and Western Blot,and separated and purified by Glutathione Sepharose4B affinity chromatography column.[Result] The pGEX-KG-Dot4 recombinant expression vector was successfully expressed in E.coli;and obtained high purified Dot4 fusion protein through purifying Glutathione Sepharose4B affinity chromatography column;Western blot analysis showed that the fusion protein interacted with GST-tagged antibody.[Conclusion]GST-Dot4 fusion protein with high purity could be obtained after it purified in E.coli,which laid a foundation of its structure,function and mechanism of Dot4p.