This study was performed to detect genetic variation of the heart fatty acid-binding protein (H-FABP) gene by PCR-RFLPs approach and its association with intramuscular fat (IMF) content. Data from 223 individuals, including one Chinese native pig breed and four western pig breeds, were analyzed. The results showed that for the H-FABP gene, there was one polymorphic Hinf site Ⅰ in the 5'-upstream region, whereas there were one HaeIII and one HinfI (marked as HinfI*) polymorphic site in the second intron, respectively. The three PCR-RFLPs were present in all breeds tested. The allele frequencies, however, revealed significant differences between them (p<0.05). Furthermore, the allele frequency distribution of HinfI in the Laiwu Black and that of HinfI* in the Hampshire breed were at disequilibrium, which might be the result of selective breeding. Results also indicated that for HinfI, HaeIII and HinfI* H-FABP RFLP, significant (p<0.05) contrasts of 0.78%, -0.69% and 0.72% were detected in the least square means of IMF content between the homozygous genotype HH and hh, DD and dd, BB and bb classes, respectively. It implied that the HHddBB genotype had the highest IMF content in this experimental population and these H-FABP RFLPs could serve, to some extent, as genetic markers for use in improvement of IMF content.