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In this study, we report the investigation of extracellular fatty acid binding protein gene ( Ex- FABP) genetic polymorphism in a sample of 360 chicken individuals. The screening of the coding regions with their intron-exon boundaries and the proximal flanking regions was performed through a PCR-SSCP strategy. Following sequence analysis revealed 35 novel single nucleotide polymorphisms (SNPs) of chicken Ex- FABP gene. Among the 35 SNPs, twenty-five were found in the introns. And the remaining seven and three SNPs were in the coding region and the 5′UTR, respectively. Two SNPs in the coding region caused two missense mutants and the other five did not result in any amino acid changes. The nature and the distribution of Ex- FABP mutations in three chicken breeds were analyzed. Variations detected here might have an impact on Ex-FABP activity and function and underpin the development of gene markers for chicken fatty deposition and metabolism. The polymorphism, generated by C4715T mutation in exon5, was significantly associated with thickness of subcutaneous fat plus skin in cocks. Subcutaneous fat plus skin of cocks was more thick in TT genotype than in CC genotype ( P < 0.05). The Ex- FABP gene could be a candidate locus or linked to a QTL that significantly affects fatty deposition and metabolism in chicken. [ABSTRACT FROM AUTHOR]