Differential production of the two most common allelic variants of beta-lactoglobulin (LG), beta-LG A and beta-LG B, has been observed using PAGE. This study evaluated 733 bp of the beta-LG promoter region and 92 bp of the first exon for possible polymorphisms using denaturing gradient gel electrophoresis and nucleotide sequence analysis. Within this region, 13 single nucleotide substitution polymorphisms were detected. Twelve polymorphisms were allele specific, and one appeared to be polymorphic only for the B allele. Several potential binding sites for transcription factors were found within the promoter sequence. This study investigated the role of the G to C transversion within a consensus binding site for activator protein-2 at position-430 bp upstream from the transcription initiation site. Using the DNase-I footprint assay, we confirmed the functional importance of this point mutation and showed different binding affinities of activator protein-2 for both alleles. We discuss the possible regulatory role of activator protein-2 in the transcriptional regulation of the beta-LG gene and propose the activator protein-2 transcription factor as a modulator of gene expression of beta-LG.