The objective of this research was to further characterize the promoter regions of the bovine and porcine fatty acid-binding protein 4 (FABP4) genes relative to those of other mammals. The DNA sequences of FABP4 promoters for the human, mouse, cow, pig, and dog were obtained from the genomic database of the National Center for Biotechnology Information and also from the sequencing of bovine and porcine genomic DNA clones obtained by 5' PCR racing of genomic DNA. Sequence alignments of these FABP4 promoters using the basic local alignment search tool of the National Center for Biotechnology Information revealed 3 highly conserved promoter regions across the species. Two computational bioinformatics databases and the literature identified the conserved transcription factor-binding sites for C/EBP, adapter primer-1, and boxes of CAAT and TATA in the first conserved proximal promoter region, a direct repeat 1-type PPAR responsive element in the second distal conserved region, and another PPAR responsive element in the third distal conserved promoter region of FABP4 in all 5 mammals. Five new short interspersed repetitive elements (SINE) in the bovine FABP4 promoter and 2 new SINE in the porcine were found, but these SINE did not disrupt the 3 conserved regions, indicating that important regulatory elements are maintained regardless of evolutionary pressure. In conclusion, the conserved cis-acting elements, especially the 2 key adipocyte transcription factors C/EBP and PPAR, may contribute greatly to adipogenic regulation and adipose tissue-specific expression of FABP4 in these mammals. This helps to further characterize and decipher important cis-acting elements that are important for adipocyte development in adipose and muscle tissue.