Abstract: The objective of the present investigation was to study genetic variations in the exon II of leptin gene in Iranian Golpayegani and Taleshi cows. Blood samples were collected from 76 Golpayegani and 64 Taleshi cows randomly selected from Delijan city (Esfahan province) and Talesh city (Guilan province) in Iran, respectively. Genomic DNA was extracted from 100 micro litters of blood using guanidium thiocyanate-silica gel method. A 94 bp fragment from exon II of the bovine leptin gene was amplified using the polymerase chain reaction. Digestion of PCR products with Bsp13I restriction enzyme differentiated C and T alleles. The T allele frequencies were 0.29 and 0.45 in Golpayegani and Taleshi breeds, respectively. The frequencies of the genotypes CC, CT and TT were 0.42, 0.58 and 0 in Golpayegani and 0.36, 0.36 and 0.27 in Taleshi cows. The average heterozygosities were 0.41 and 0.50 in Golpayegani and Taleshi breeds, respectively. The χ2 test did not confirm the Hardy-Weinberg equilibrium in both populations.