Expression and Purification of PEP-1-SOD1 Fusion Protein

Objective To construct prokaryotic expression vector pET15b-PEP-1-SOD1 and express and purify PEP-1-SOD1 fusion protein.Methods The SOD1 cDNA was amplified by PCR using plasmid pBluscript Ⅱ SK-SOD1 as a template,identified by restriction analysis and used for construction of prokaryotic expression vectors pET15b-SOD1 and pET15b-PEP-1-SOD1.The constructed recombinant plasmids were identified by sequencing and transformed to E.coli BL21(DE3) for expression of SOD1 protein and PEP-1-SOD1 fusion protein.The expressed products were identified by SDS-PAGE and Western blot.Results SOD1 and PEP-1-SOD1 proteins,with relative molecular weights of 22 000 and 26 000 respectively,were expressed.The expressed product existed in a nature and soluble form and contained 30% of total somatic protein.Conclusion SOD1 protein and PEP-1-SOD1 fusion protein were successfully prepared.