Sığır Kappa-Kazeinin A ve B Genetik Varyantlarının PCR-RFLP Analizi için Yeni Bir Primer Dizaynı

Authors : Mevlut ARSLAN

Pages : 6-11

Doi:10.31196/huvfd.651821

View : 27 | Download : 15

Publication Date : 2020-06-25

Article Type : Research Paper

Abstract :Bovine milk contains about 3.4% high-quality protein and it is very essential for human nutrition. Kappa (κ) casein is one of the milk proteins and encoded by the CSN3 gene. Studies have shown that κ-casein has an important influence on milk properties and the manufacturing of milk. κ-casein takes role as a stabilizing factor during the curdling of milk, which makes it desired in the cheese factory. A and B genetic variants of κ-casein are well-known and well-studied. For the genotyping of bovines, the PCR-RFLP approach has been used. For this aim, different primer pairs have been used to amplify the polymorphic region of the CSN3 gene. In a previous study, the HinfI enzyme digestion of the polymorphic region resulted in short and similar length fragments. In agarose gel electrophoresis, separation of very similar DNA fragments is almost impossible and interpretation of the short DNA fragment sometimes causes challenges. Therefore, in this study, a new primer design was described. Using the new primer, longer DNA fragment was amplified successfully and HinfI digestion of the PCR product let to a longer and very different lengths of DNA fragments, which can be easily separated and interpreted in agarose gel electrophoresis. The primer described in this study can be used in further studies related to allele frequency researches and breeding strategies.
Keywords : Milk, κ casein, CSN3, Polymorphism, PCR RFLP, Primer design