Genotype frequencies in calpastatin (CAST) and callipyge (CLPG) genes in Northeast Bulgarian Merino sheep breed using PCR-RFLP method

摘要

The purpose of the present work was to identify polymorphic allelic variants of ovine calpastatin and callipyge genes in Northeast Bulgarian Merino sheep breed. Blood samples were collected from 32 rams, reared in Agricultural Experimental Station - Targovishte. Genomic DNA was extracted using commercial kit for DNA purification from whole blood. After PCR amplification of CAST gene with specific primer set a fragment of 622 bp was obtained. CLPG gene also was amplified with specific primers and a fragment of 426 bp was obtained. Genotypes were determined by restriction fragment length polymorphism (RFLP) method with MspI restriction enzyme for CAST gene and BsmFl restriction enzyme for CLPG gene. Calpastatin locus was found to be polymorphic, whereas in callipyge gene it was observed uniformity - the presence of genotype AA only. Genotype frequencies established in CAST gene were 0.47% and 0.53% for MM and MN, respectively, whereas genotype NN was not found.