In order to investigate the effect of ADFP gene on the carcass traits and intramuscular fat content of Da-heng high-quality chicken, we analyzed the coding sequence of the ADFP gene by single-strand conformation polymorphism (SSCP) and DNA sequencing in five pure lines and three crossbreds. We found three single nucleotide polymorphism sites, including A/T at base position 4,079 (locus A), C/T at base position 4,843 (locus B) and A/G at base position 7,070 (locus C). GLM analysis for the effect of genotypes on carcass traits and intramuscular fat content demonstrated a significant effect on leg muscle percentage, abdomen fat weight, abdominal fat percentage and intra-muscular fat content in locus A (P < 0.05), a significant association with living body weight and carcass weight for locus B (P < 0.05) and a significant association with breast muscle weight and intramuscular fat content (P < 0.05), and a very significant association with breast muscle percentage for locus C (P < 0.01). It was concluded that ADFP gene may be a major gene or linked to such a gene that affects the fat traits of birds. Polymorphic loci A and C could be used as molecular markers for abdominal fat weight, abdominal fat percentage and intramuscular fat content in birds in future studies.