Abstract: Objective To construct the expression vector pcDNA3.1/myc-his-DJ-1 and pcDNA3.1/myc-His-DJ-1^M26I to lay a foundation of further study on function of DJ-1^M26I and establish transgenic animals. Methods The 26^th amino acid of DJ-1 was mutated by gene mutation kit, then pcDNA3.1/myc-his-DJ-1 and pcDNA3.1/myc-His-DJ-1^M26I were constructed. The two plasmids were transfected into NIH3T3 cells respectively using liposome method, then the cells were selected with G418 and detected the DJ-1 gene expression at DNA, transcription and protein levels. Results 1 clone of pcDNA3.1/myc-his-DJ-1 and 3 clones of pcDNA3.1/myc-his-DJ-1^M26I were obtained after G418 selection by PCR analysis. The expression of DJ-1-His were detected by RT-PCR and Western blot in pcDNA3.1/myc-his-DJ-1 and pcDNA3.1/myc-his-DJ-1^M26I transfected cells. The profieration rate of DJ-1^M26I transfected cells was lower than normal NIH3T3 cells (P<0.05) by MTT essay. Conclusion The expression vectors pcDNA3.1/myc-his-DJ-1 and pcDNA3.1/myc-His-DJ-1^M26I were constructed successfully.

Key words: DJ-1, NIH3T3 cell, Parkinson's disease