The genotypes for Calpastatin, K-Casein, Leptin and BoLA-DRB3 Loci were determined by polymerase chain reaction and restriction enzyme digestion method in native Iranian breed cattle, Sistani. Blood samples were collected from Sistani breeding station located in Zehak, Zabol. The extraction of genomic DNA was based on Guanidin Thiocyanate-Silica gel method. After PCR reaction, amplicons were digested with restriction enzymes. The Calpastatin locus had 3 genotypes with frequencies of 0.62, 0.29 and 0.09 for MM, MN and NN, respectively; K-Casein and Leptin had 3 genotypes with frequencies of 0.27, 0.57 and 0.16 for K-Casein, 0.77, 0.22 and 0.01 for Leptin for AA, AB and BB genotypes, respectively. For BoLA-DRB3 we identified 19 alleles and DRB3.2*8 had the highest allelic frequency -22.4%-. One of the 19 allele had a new pattern. Average heterozygosity value for all loci was low. χ2 test did not confirm the Hardy-Weinberg equilibrium for Calpastatin in this population. These data provide evidence that Iranian’s Sistani breed have a variability, which opens interesting prospects for future selection programs, especially marker-assistant selection.

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