Determination of beta lactoglobulin and calpastatin gene polymorphism by PCR-RFLP method in akkaraman sheep in Yüksekova district of Hakkari province

This study was carried out to determine beta lactoglobulin (BLG) and calpastatin (CAST) gene polymorphism in Akkaraman sheep by PCR-RFLP method. The material of the study consisted of 100 Akkaraman ewes aged between 2 and 5 years, which were raised in a private sheep farming enterprise operating in Yüksekova district of Hakkari province. A total of 9 ml of blood was taken from the vein of the sheep's neck into tubes with EDTA. DNA samples were isolated from the blood samples using commercial ready-made kits. The target DNA region of BLG and CAST genes were amplified by PCR. The PCR products were cut with RsaI and MspI restriction enzymes to determine the genotypes of BLG and CAST genes, respectively. The restriction fragments were run on 2% agarose gel electrophoresis. Fragments were visualised under UV light. Gene and genotype frequencies were calculated by direct counting method. Chi-square (χ2) test was used to check whether the population was in Hardy-Weinberg equilibrium. In the study, the frequencies of AA, AB and BB genotypes of BLG gene were found to be 23.0, 68.0 and 9.0 %, respectively, and the frequencies of A and B alleles were 57.0 and 43.0 %, respectively. The frequencies of MM, MN and NN genotypes of CAST gene were 61.0, 34.0 and 5.0 %, respectively, and the frequencies of M and N alleles were 78.0 and 22.0 %, respectively. It was observed that Akkaraman sheep population was in Hardy-Weinberg equilibrium in terms of CAST gene, but not in equilibrium in terms of BLG gene. As a result, it was observed that heterozygous AB genotype of BLG gene was more common in Akkaraman sheep population and homozygous MM gene was more common in CAST gene. The distribution of BLG and CAST genes within the Akkaraman sheep population revealed that this breed has significant breeding potential. Key words: Akkaraman Sheep, Beta Lactoglobulin, Calpastatin, PCR-RFLP.