Objective    To bioinformatically analyze the human NaDC1 promoter for predicting possible regulative zone and cis element and construct firefly luciferase report gene vector for 5’ flanking regulated elements of hNaDC1 gene.     Methods    The proximal promoter of hNaDC1 was analyzed by using FirstEF online software to obtain its sequence. Meanwhile

the binding sites of transcription factors were predicted by software Matinspector 5.0. The DNA fragment hNaDC1A (-2 232/+136) of 5’ flanking of hNaDC1 gene were amplified from the genomic DNAs of human renal proximal tubular cells (HK-2) by PCR

the products of which were cloned into pGL3-Basic after digestion with KpnⅠ and Hind Ⅲ. The recombinant plasmid pGL3-hNaDC1A was used for restriction endonucleases digestion and DNA sequencing.     Results    The 2.4 kb (-2 232/+136) sequence of hNaDC1 proximal promoter was obtained by FirstEF. Matinspector 5.0 showed 152 cis-acting elements (74 sorts) within the proximal promoter. The expression vector pGL3-hNaDC1A was constructed successfully.     Conclusion    The construction of firefly report gene vector offers experimental basis for studying the distribution of transcriptional regulation elements and their characteristics with the help of dual luciferase report gene assay system.