Polymorphism of Calpastatin Gene in Crossbreed Dalagh Sheep Using PCR-RFLP

Abstract

In modern programs of animal breeding, the genetic polymorphisms of production traits can be used as marker systems. Calpastatin gene is located on ovine chromosome 5 and is polymorphic in many breeds of sheep. Calpastatin has a role in meat tenderness after slaughter. Blood samples were collected from 120 crossbreeds of Dalagh sheep located in Golistan province of Iran. Genomic DNA was extracted from blood sample. Gel monitoring and spectrophotometer methods were used to determine the quality and quantity of DNA. Intron I from L domain of the ovine calpastatin gene was amplified to produce a 565 bp fragment. The PCR products were digested by restriction endonucleases MspI. Digested products were separated by electrophoresis on 1.5% agarose gel and visualized after staining with ethidium bromide on UV transillumination. The MspI digestion of the PCR products produced digestion fragments of 306 and 259 bp. Data analysis was done using PopGen32 software. In this population, MM, MN and NN genotype were identified with 65.5, 29, 5.5% frequencies, respectively. The population was found to follow Hardy-Weinberg equilibrium.

Key words: Calpastatin gene, polymorphism, crossbreed, Dalagh sheep.