A Novel Test System for Genotyping rs43703016 Single-nucleotide Substitutions in the Bovine CSN3 Gene
Svetlana Kovalchuk
Center of Experimental Embryology and Reproductive Biotechnologies, Moscow, Russian Federation.
Arina Tagmazyan
Lomonosov Moscow State University, Faculty of Biology, Moscow, Russian Federation.
Eugene Klimov *
Center of Experimental Embryology and Reproductive Biotechnologies, Moscow, Russian Federation and Lomonosov Moscow State University, Faculty of Biology, Moscow, Russian Federation.
*Author to whom correspondence should be addressed.
Abstract
Aims: Caseins are among the main milk proteins that determine many of its properties. Bovine kappa-casein (CSN3) is associated with the qualitative composition of milk, as well as with the quality of cheese obtained from this milk. The rs43703016 single-nucleotide substitution (g.88532332A>C; Asp148Ala) in exon 4 of the bovine CSN3 gene plays an important role in the production of quality hard cheeses. Various methods for the DNA testing of this substitution have been developed in the last three decades. Emergent DNA technologies provide an opportunity to modernize methods of genotyping single-nucleotide polymorphisms.
Results: We have developed and verified a method to differentiate A/C alleles of the rs43703016 substitution in the bovine CSN3 gene by real-time PCR using allele-specific fluorescent probes.
Conclusion: Our new method allows fast genotyping of animals, and may be used for selection of cows carrying the CC genotype, which determines good cheese-making properties of milk.
Keywords: Cattle, CSN3 gene, rs43703016 single-nucleotide, genotyping, real-time PCR.