Polymorphism investigation of calpastatin gene in Zel sheep population of Iran by PCR-RFLP method

Abstract

Meat tenderness is an importantquality characteristicfor which consumersare interested. Calpastatin is the calpain inhibitor enzyme and plays an important role in muscle growth and meat quality. The calpastin gene is located on sheep chromosome 5 and its polymorphisms are associated with economic traits. This study was performed to identify calpastatin gene polymorphisms using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. Random blood samples were collected from 100 Zel sheep. DNA was extracted by the salting out method and was amplified using PCR. The quantity and quality of the extracted DNA were determined by spectrophotometry and agarose gel electrophoresis. For amplification, a 622 bp fragment exon 1 of the calpastatin gene was used. Amplified fragments were digested by Msp1 restriction enzyme to determine the calpastatin genotypes, and two alleles. M and N were identified. The genotype frequencies of the MM, MN and NN were 0.62, 0.26 and 0.12, respectively in the population under study. Also, the allelic frequency of M and N were 0.75 and 0.25, respectively. Investigation of population genetic structure of Zel sheep revealed that it was not in Hardy-Weinberg equilibrium (p<0.05). The results indicate that it could be useful to consider genetic diversity at calpastatin locus in Zel sheep.

Key words: Calpastatin gene, polymorphism, Zel sheep, meat and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP).