Abstract: The mutations or loss of DJ-1 gene cause Parkinson’s disease（PD）, but the role of DJ-1 and its relative mechanisms for PD are still under study and investigation. Dictyostelium discoideum is one of the recognized models for study of neurodegenerative disease. To study the role of DJ-1 in PD and its mechanisms, the specific primers were designed to amplify the full-length Dictyostelium discoideum DJ-1 and its fragment from 75～479 bp using two-step cloning strategy. The DNA of interest and pUC18 vector were digested and ligated with T4 ligase. The product of ligation was transformed into Escherichia coli DH5α and followed by screening of strains of interest using Blue/white selection and gene sequencing. The D. discoideum prokaryotic expression construct pPROF681 and pPROF682 were built and verified by endonuclease digestion and gene sequencing. The genes of interest were then cut by SacⅠ, Bam HⅠ, Eco RⅠ and XhoⅠ designed in primers from pPROF681 and pPROF682 and inserted into D. discoideum vectors p DNeo2 and pPROF267 to create D. discoideum eukaryotic expression vectors pPROF688 and p PROF690. This work provides a very important basis for further achievement of D. discoideum strains con-taining pPROF688 and pPROF690, selection and verification of interested transformants, analysis of the relativities between expression level of DJ-1 and the phenotypes of D. discoideum and built of D. discoideum model of DJ-1 for PD.