Objective:To construct luciferase reporter plasmids of full-length and truncated promotors of rat response gene to complement 32 (RGC32) gene and detect their activity in HEK293 cells in response to myeloid zinc finger gene 1 (MZF1) overexpression,screening the possible binding sites for MZF1. Methods:Rat RGC32 promoter(-686~-1 nt) was amplified by PCR and cloned into the luciferase reporter plasmid (pGL3-basic). The recombinant plasmid (pGL3-RGC32-FL) and rat MZF1 expression plasmid (pIRES2-EGFP-MZF1) were co-transfected into HEK-293 cells and then the luciferase activity was detected to determine the role of MZF1 in RGC32 gene transcription. Meanwhile,the potential MZF1 binding sites within RGC32 promoter were predicted by bioinformatics software. Based on the predicted results,different luciferase reporter plasmid of truncated MZF1 gene promotor that named pGL3-RGC32-1,pGL3-RGC32-2 and pGL3-RGC32-3 were constructed. The promoter luciferase reporter plasmids of pGL3-RGC32-FL or pGL3-RGC32-1,pGL3-RGC32-2,pGL3-RGC32-3 and the plasmid of pIRES2-EGFP-MZF1 were co-transfected into HEK293 cells. Then,the luciferase activity was detected to screen the MZF1 binding sites. Results:It was verified that different kinds of plasmids were all constructed correctly by PCR analysis and nucleotide sequencing. The plasmids of pGL3-RGC32-FL and pIRES2-EGFP-MZF1 were also co-transfected into HEK293 cells,and then the luciferase activity was detected. The results showed that the transcriptional activity of RGC32 gene was increased markedly in response to MZF1 overexpression. In addition,the plasmids of pGL3-RGC32-FL or pGL3-RGC32-1,pGL3-RGC32-2 and pGL3-RGC32-3 and pIRES2-EGFP-MZF1 were co-transfected into HEK293 cells,and then the luciferase activity in different groups was determined. The result displayed that the activity of pGL3-RGC32-3 was much lower than that in pGL3-RGC32-FL,pGL3-RGC32-1 and pGL3-RGC32-3,indicating that the region of rat RGC32 promoter (-286~-86 nt) might contain MZF1 binding element. Conclusion:The rat full-length and truncated rat RGC32 promotor luciferase reporter plasmids were constructed successfully,overexpress MZF1 could increase the transcriptional activity of RGC32 gene,and the MZF1 binding region was identified.