Here we report the cloning and characterization of chicken visfatin (also called pre-B cell enhancing factor; PBEF, or nicotinamide phosphoribosyltransferase; Nampt) gene. Sequence analyses revealed that the coding region of visfatin is 1,482 bp in length and encodes a protein of 493 amino acids, which shares high amino acid sequence identity not only to visfatin of human (94%), rat (94%), carp (89%), and zebrafish (89%), but also to Nampt of sponge (58%) and cyanobacterium (48%). The reverse transcription PCR assay and Northern-blot analysis demonstrated that visfatin was widely expressed in all chicken tissues examined. Using a dual luciferase reporter system, we further demonstrated that the cloned 1,372-bp fragment upstream of the putative translation start site (ATG) displayed the maximal promoter activity in cultured CHO, DF-1, and HEK293 cells, whereas the removal of its 5'-region (1,075 bp) or 3'-region (297 bp) could only partially reduce its promoter activity, implying that visfatin gene transcription was likely controlled by multiple promoters near the translation start site. Taken together, results from present study will contribute to our better understanding of the expression and roles of visfatin gene in chickens.