We have identified the functional promoter region of the human phospholipid transfer protein gene. Primer extension analysis indicates multiple sites for transcription initiation. Sequence analysis reveals that the promoter consists of TATA box, high GC region, and several consensus sequences for the potential binding of transcription factors. To assay promoter activity, DNA fragments with various lengths of the 5'-flanking region were fused upstream to the luciferase gene and transfected into HepG2, COS-7, and CHO cells. A minimal promoter of 159 base pairs between -230 and -72 relative to the first transcriptional initiation site is responsible for the full activity. Two motifs, Sp1 and AP-2, are located within this area. It may suggest that the PLTP promoter activity relies primarily on the putative cis-elements in the functional region.