Calpastatin has been known as candidate gene in muscle growth efficiency and meat quality. This gene has been located to chromosome 5 of sheep. In order to evaluate the calpastatin gene polymorphism, random blood sample were collected from 111 Arabic ram sheep from different regions. The DNA extraction was based on Boom et al. (1989) method. Exon and entron I from L domain of the ovine calpastatin gene was amplified to produce a 622 bp fragment. The PCR products were electrophoresed on 1.2% agarose gel and stained by etidium bromide. Then, they were digested with restriction enzyme MspI and then electrophoresed on 2.5% agarose gel with ethidium bromide and revealed two alleles, allele A and allele B. Data were analysed using PopGene32 package. In this population, AA, AB, BB genotype have been identified with the 70.27, 28.82, 0.9% frequencies. A and B allele’s frequencies were 0.85, 0.15, respectively. The population was found to follow Hardy-Weinberg equilibrium.